Yrd. Doç. Dr.

Suzan BİRAN AY
Microbial Growth

• an increase in cell number, not in cell size!

• from 1 mother cell 2 daughter cells are formed.

• most bacterial cells reproduce asexually by binary fission. This

involves several stages:
o increasing cell size (growth)
o DNA replication, and
o division (septum formation)
Microbial Growth
Microbial Growth

• Generation Time: Time required for a cell population to

double
DNA

DNA Replication

12 Cell Elongation
9 3
Septum Formation
6

Cell Separation
 Microbial Growth

• Generation time varies considerably:

• Depends on environmental conditions
• Depends on microbial type
o E. coli divides every 20 minutes.
o Most bacteria divide every 1 to 3 hours.
o Some bacteria require over 24 hours to
divide.
• We can express the number of cells in a
bacterial generation as 2n, where n is the
number of doublings that have occurred.
 Microbial Growth

• Bacterial Growth Curve: When bacteria

are inoculated into a liquid growth
medium, we can plot of the number of
cells in the population over time.
• Phases of growth (Batch fermentation)
1. Lag phase
2. Exponential (logarithmic [log])
phase )
3. Stationary phase
4. Death phase
 Microbial Growth

1. Lag Phase:
• Period of adjustment to new conditions.
• Little or no cell division occurs,
population size doesn’t increase.
• Phase of intense metabolic activity, in
which individual organisms grow in size;
usually by synthesizing new cell
components (ATP, Ribosomal proteins, rRNA,
tRNA,Co-factors,Enzymes)
• May last from one hour to several days
Microbial Growth

2. Log Phase:
• Cells begin to divide and generation
time reaches a constant minimum.
• Period of most rapid growth.
• Number of cells produced > Number
of cells dying
• Cells are at highest metabolic activity.
• Cells are most susceptible to adverse
environmental factors at this stage.
•Radiation
•Antibiotics
Microbial Growth

2. Log Phase:
• Calculation of generation time:
• If 100 cells growing for 5 hours produced 1,720,320

𝒍𝒐𝒈 𝒆𝒏𝒅 𝒏𝒖𝒎𝒃𝒆𝒓 − 𝒍𝒐𝒈(𝒊𝒏𝒊𝒕𝒊𝒂𝒍 𝒏𝒖𝒎𝒃𝒆𝒓)

𝑵𝒖𝒎𝒃𝒆𝒓 𝒐𝒇 𝒈𝒆𝒏𝒆𝒓𝒂𝒕𝒊𝒐𝒏𝒔 =
𝒍𝒐𝒈𝟐

log 1720320 − log(100)

𝑁𝑢𝑚𝑏𝑒𝑟 𝑜𝑓 𝑔𝑒𝑛𝑒𝑟𝑎𝑡𝑖𝑜𝑛𝑠 = = 14.07
0,301

𝟔𝟎 𝒎𝒊𝒏 × 𝒉𝒐𝒖𝒓𝒔
𝑮𝒆𝒏𝒆𝒓𝒂𝒕𝒊𝒐𝒏 𝒕𝒊𝒎𝒆 =
𝒏𝒖𝒎𝒃𝒆𝒓 𝒐𝒇 𝒈𝒆𝒏𝒆𝒓𝒂𝒕𝒊𝒐𝒏𝒔
60 𝑚𝑖𝑛 × 5
𝐺𝑒𝑛𝑒𝑟𝑎𝑡𝑖𝑜𝑛 𝑡𝑖𝑚𝑒 = = 21 𝑚𝑖𝑛𝑢𝑡𝑒𝑠/𝑔𝑒𝑛𝑒𝑟𝑎𝑡𝑖𝑜𝑛
14.07
Microbial Growth

3. Stationary Phase:
• Population size begins to stabilize.
• Number of cells produced = Number of
cells dying
• Overall cell number does not increase.
• Cell division begins to slow down.
• Factors that slow down microbial
growth:
o Accumulation of toxic waste materials
o Acidic pH of media
o Limited nutrients
o Insufficient oxygen supply
Microbial Growth

4. Death or Decline Phase:

• Population size begins to
decrease.
• Number of cells dying >
Number of cells produced
• Cell number decreases at a
logarithmic rate.
• Cells lose their ability to divide.
• A few cells may remain alive for
a long period of time.
Microbial Growth

1. Batch culture (closed system)

2. Semi-continuous (Fed-batch)
3. Continuous (open system)
Microbial Growth

1. Batch culture (closed system)

• a closed-system microbial culture of
fixed volume
• in each batch culture there are:
o cleaning
o fill-up
o sterilization
o fermentation
o emptying
Microbial Growth

2. Fed-Batch culture
• At t=0 almost all substrates are added,
fermentor strilized and microbial culture
put in the reactor
• One of the substrates is not added in high
concentration because it may be toxic or as
in yeast its addition should be slow
Microbial Growth

3. Continuous culture
• Used to maintain cells in the exponential growth phase at a
constant biomass concentration for extended periods of time
• Conditions are met by continual provision of nutrients and
removal of wastes = OPEN SYSTEM
• Constant conditions are maintained
Microbial Growth

3. Continuous culture
A. Chemostat
• A continuous culture device that maintains a
constant growth rate by:
o supplying a medium containing a limited
amount of an essential nutrient at a fixed
rate
o removing medium that contains
microorganisms at the same rate
• As fresh media is added to the chamber,
bacteria are removed
• Limiting nutrients control growth rates
• Cell density depends on nutrient concentration
Microbial Growth

3. Continuous culture
B. Turbidostat
• A continuous culture device that regulates the
flow rate of media through the vessel in order
to maintain a predetermined turbidity or cell
density
o There is no limiting nutrient
o Absorbance is measured by a photocell
(optical sensing device)
o The number of cells in culture controls
the flow rate and the rate of growth of
culture adjusts to this flow rate
Microbial Growth

1. Physical
a) Temperature
b) pH
c) Osmotic pressure
2. Chemical
a) Carbon
b) Nitrogen, sulfur, phosphorous
c) Trace elements
d) Oxygen
 Microbial Growth

• Microorganisms are sensitive to

temperature changes
o Enzymes have temperature optima
o If temperature is too high, proteins
denature, including enzymes,
carriers and structural components
• Temperature ranges are
enormous (-20 to 100oC)
Microbial Growth

Microbes are loosely classified into several groups based on their

preferred temperature ranges.
A. Psychrophile: low temperature
B. Mesophile: midrange temperature
C. Thermophile: high temperature
D. Hyperthermophile (extreme thermophile): very
high temperature
Microbial Growth

Microbes are loosely classified into several groups based on their

preferred temperature ranges.
A. Psychrophiles: “Cold-loving”. Can grow at 0oC. Two
groups:
o True Psychrophiles: Sensitive to temperatures over
20oC. Optimum growth at 15oC or below. Found in very
cold environments (North pole, ocean depths).
Seldom cause disease or food spoilage.
o Psychrotrophs: Optimum growth at 20 to 30oC.
Responsible for most low temperature food spoilage.
Microbial Growth

B. Mesophiles: “Middle loving”. Most bacteria.

o Include most pathogens and common spoilage organisms.
o Best growth between 25 to 40oC.
o Optimum temperature commonly 37oC.
o Many have adapted to live in the bodies of animals.
Microbial Growth

C. Thermophiles: “Heat loving”.

o Optimum growth between 50 to 60oC.
o Many cannot grow below 45oC.
o Adapted to live in sunlit soil, compost piles, and hot springs.
o Some thermophiles form extremely heat resistant
endospores.
D. Extreme Thermophiles (Hyperthermophiles): Optimum
growth at 80oC or higher. Archaebacteria. Most live in volcanic
and ocean vents.
Microbial Growth

• Growth Rates of Bacterial Groups at Different Temperatures

Microbial Growth

• Most bacteria prefer neutral pH (6.5-7.5).

• Molds and yeast grow in wider pH range, but prefer pH
between 5 and 6.
• Acidity inhibits most microbial growth and is used frequently
for food preservation (e.g.: pickling).
• Alkalinity inhibits microbial growth, but not commonly used for
food preservation.
• Acidic products of bacterial metabolism interfere with growth.
Buffers can be used to stabilize pH.
Microbial Growth

Organisms can be classified as:

A. Acidophiles: “Acid loving”.
o Grow at very low pH (0.1 to 5.4)
o Lactobacillus produces lactic acid, tolerates mild acidity.
B. Neutrophiles:
o Grow at pH 5.4 to 8.5.
o Includes most human pathogens.
C. Alkaliphiles: “Alkali loving”.
o Grow at alkaline or high pH (7 to 12 or higher)
Microbial Growth

• Microbes require water to dissolve enzymes and nutrients

required in metabolism
• Water is important reactant in many metabolic reactions
• Most cells die in absence of water
– Some have cell walls that retain water
– Endospores and cysts cease most metabolic activity in a dry environment
for years
• Two physical effects of water
– Osmotic pressure
– Hydrostatic pressure
 Microbial Growth

• Is the pressure The

exerted on a solution … the height
semipermeable increases of solution
in volume exerts the
membrane by a
until … osmotic
solution containing
pressure (π)
solutes that cannot of the
freely cross solution.
membrane; related
to concentration of
dissolved
molecules and ions
in a solution Net flow of water from
the outside (pure H2O)
to the solution.
Microbial Growth

• Water is solvent for biomolecules, and its availability is critical

for cellular growth
• The availability of water depends upon its presence in the
atmosphere (relative humidity) or its presence in solution or a
substance (water activity, (aw))
• aw of pure water is 1.0; affected by dissolved solutes such as
salts or sugars.
• aw is inversely related to osmotic pressure
o If the solution has a high osmotic pressure (high
extracellular solute concentration), then its aw = low
Microbial Growth

Cells are 80 to 90% water.

A. Hypertonic solutions: High osmotic pressure removes water
from cell, causing shrinkage of cell membrane (plasmolysis).
o Used to control spoilage and microbial growth.
 Sugar in jelly
 Salt on meat
B. Hypotonic solutions: Low osmotic pressure causes water to
enter the cell. In most cases cell wall prevents excessive entry of
water. Microbe may lyse or burst if cell wall is weak (osmotic
lysis).
Microbial Growth

• Effects of Osmosis on
Bacterial Cells
Microbial Growth

Classification:
A. Halophiles: Require moderate to large salt concentrations.
Ocean water contains 3.5% salt.
o Most bacteria in oceans.
B. Extreme or Obligate Halophiles: Require very high salt
concentrations (20 to 30%).
o Bacteria in Dead Sea, brine vats.
C. Facultative Halophiles (or Halotolerants): Do not require high
salt concentrations for growth, but tolerate 2% salt or more.
Microbial Growth

•
Microbial Growth

• Water exerts pressure in proportion to its depth

– For every addition of depth, water pressure increases
1 atm
• Organisms that live under extreme pressure are barophiles
– Their membranes and enzymes depend on this pressure to
maintain their three-dimensional, functional shape
Will continue next week