Chapter 7

The Control of Microbial

Growth
Dr. Ayman El-Shibiny
aelshibiny@zewailcity.edu.eg

Objectives
Learn the terminology of microbial control
Study the physical methods of microbial control
Heat
Filtration
Low temperature
High pressure
Desiccation
Radiation

Objectives
Learn the terminology of microbial control
Study the physical methods of microbial control
Heat
Filtration
Low temperature
High pressure
Desiccation
Radiation

The Terminology of Microbial Control

Sterilization: the removal or destruction of all
living microorganisms
Sterilant : A sterilizing agent
Commercial sterilization: killing C. botulinum
endospores
Disinfection: the destruction of vegetative,
non-endospore forming, pathogens
Antisepsis: the destruction of vegetative
pathogens on living tissue
Antiseptic: the chemical used for antisepsis

The Terminology of Microbial Control

Degerming (Degermation): removing
microbes from a limited area
Sanitization: lowering microbial counts on
eating utensils
Biocide/germicide: killing microbes
Bacteriostasis: inhibiting, not killing, microbes

The Terminology of Microbial Control

Sepsis refers to microbial contamination, as in
septic tanks for sewage treatment.
Asepsis is the absence of significant
contamination
Aseptic surgery techniques prevent microbial
contamination of wounds

The Rate of Microbial Death

When bacterial populations are heated or
treated with antimicrobial chemicals, they
usually die at a constant rate.

Microbial Exponential Death Rate: An Example

Understanding the Microbial Death Curve.

One log decrease =

90% of population
killed

Arithmetic number of surviving cells

log10 of number of surviving cells

Plotting the typical microbial death curve

logarithmically (red line) results in a straight line.

Time (min)

(a) Plotting the typical microbial death curve arithmetically

(blue line) is impractical: at 3 minutes the population of
1000 cells would only be a hundredth of the graphed
distance between 100,000 and the baseline.

Effectiveness of Treatment
Depends on:
Number of microbes
Environment (organic matter, temperature,
biofilms). In suspending medium, fats and proteins
protect the microbes
Because their activity is due to temperaturedependent chemical reactions, disinfectants work
somewhat better under warm conditions.
Time of exposure
Microbial characteristics

Understanding the Microbial Death Curve.

log10 of number of surviving cells

sterile surgical equipment

Time (min)

(b) Logarithmic plotting (red) reveals that if the

rate of killing is the same, it will take longer to kill
all members of a larger population than a smaller
one, whether using heat or chemical treatments.

Actions of Microbial Control Agents

Alteration of membrane permeability (plasma

membrane)
Damage to proteins and lipids of the plasma
membrane by antimicrobial agents
After damage, cellular contents leak into the
surrounding medium
Destroy the hydrogen bonds which link the
amino acids of enzymes (proteins) by heat or
certain chemicals
Damage to nucleic acids by heat, radiation, or
chemicals is frequently lethal to the cell

Objectives
Learn the terminology of microbial control
Study the physical methods of microbial control
Heat
Filtration
Low temperature
High pressure
Desiccation
Radiation

Physical Methods of Microbial Control

Drying (desiccation) and salting at the stone
age
The best method to control microbes?
1. Heat might inactive vitamins or antibiotics in
a solution
2. Repeating heat may damage many laboratory
and hospital materials such as rubber and
latex tubing

Heat
Heat: Canned foods, laboratory media and
glassware, and hospital instruments
Heat denatures enzymes
Thermal death point (TDP): lowest
temperature at which all cells in a culture are
killed in 10 min
Thermal death time (TDT): time during which
all cells in a culture are killed
Decimal Reduction Time (DRT) or D value:
Minutes to kill 90% of a population at a given
temperature

Understanding the Microbial Death Curve.

One log decrease =

90% of population
killed

Arithmetic number of surviving cells

log10 of number of surviving cells

Plotting the typical microbial death curve

logarithmically (red line) results in a straight line.

Time (min)

(a) Plotting the typical microbial death curve arithmetically

(blue line) is impractical: at 3 minutes the population of
1000 cells would only be a hundredth of the graphed
distance between 100,000 and the baseline.

Moist Heat Sterilization

Moist heat denatures (coagulates) proteins by
breaking the hydrogen bonds that hold the
protein in their three dimensional structure.
Sterilization, a type of mist heat, is boiling
which kills vegetative cells of bacterial
pathogens in 10 about minutes
Autoclave: steam under pressure
The higher the pressure in the autoclave, the
higher the temperature
In autoclave, Steam must contact items surface

An autoclave.
Exhaust valve
(removes steam
after sterilization)

Steam to
chamber

Safety
valve

Pressure gauge

Operating valve
(controls steam from
jacket to chamber)

Steam

Door
Steam
chamber

Air

Perforated shelf
Steam jacket

To waste line

Sediment
screen
Thermometer

Automatic ejector valve

Pressure regulator
(thermostatically controlled;
for steam supply
closes on contact with
pure steam when air is
Steam supply
exhausted)

Examples of sterilization indicators.

Pasteurization

Reduces spoilage organisms and pathogens

Different products have different pasteurization
times and temperatures.
Phosphatase test, in milk, to test pasteurization
Equivalent treatments
63C for 30 min
High-temperature short-time (HTST): 72C for 15
sec. to store milk under refrigeration condition
Ultra-high-temperature (UHT): 140C for <1 sec.
To store the product for several months without
refrigeration

 Many relatively heat-resistant (thermoduric)

bacteria survive pasteurization, but these are
unlikely to cause disease or cause refrigerated
milk to spoil.
Equivalent treatments (140C for 4 seconds ):
as the temperature is increased, much less time
is needed to kill the same number of microbes
The destruction of highly resistant endospores
might take 70 minutes at 115C, whereas only
7 minutes might be needed at 125C

Dry Heat Sterilization

Kills by oxidation
Dry heat
Flaming
Incineration
Hot-air sterilization

Equivalent Treatments

Hot-Air

Autoclave

170C, 2 hr

121C, 15 min

Objectives
Learn the terminology of microbial control
Study the physical methods of microbial control
Heat
Filtration
Low temperature
High pressure
Desiccation
Radiation

Filtration
Is the passage of a liquid or gas through a
screenlike material with pores small enough to
retain microorganisms.
It is used to sterilize heat-sensitive materials
High-efficiency particulate air (HEPA) removes
microbes >0.3 m
Membrane filtration removes microbes >0.22
m

Filter sterilization with a disposable, presterilized plastic unit.

Flask of
sample
Cap

Membrane filter

Cotton plug in
vacuum line
ensures sterility
Sterile
filtrate

Vacuum line

Objectives
Learn the terminology of microbial control
Study the physical methods of microbial control
Heat
Filtration
Low temperature
High pressure
Desiccation
Radiation

Physical Methods of Microbial Control

The effect of low temperatures on
microorganisms depends on the particular
microbe and the intensity of application
Low temperature inhibits microbial growth
Refrigeration: the metabolic rate is reduced that
microbes cannot reproduce or synthesis toxins
Pathogenic bacteria generally will not grow at
refrigeration tempreature

 Deep-freezing: do not necessary kill the bacteria

Slow freezing is more harmful to bacteria
Once frozen, one third of the population of some
vegetative bacteria might survive a year
Lyophilization

Objectives
Learn the terminology of microbial control
Study the physical methods of microbial control
Heat
Filtration
Low temperature
High pressure
Desiccation
Radiation

 High pressure denatures proteins: if the

pressure is high enough, it alters the
molecular structures of proteins and
carbohydrates
Endospore is relatively resistant to high
pressure
Can be used in food industry

Objectives
Learn the terminology of microbial control
Study the physical methods of microbial control
Heat
Filtration
Low temperature
High pressure
Desiccation
Radiation

 Desiccation (lyophilization) prevents

metabolism: microorganisms can remain
viable for years. When water is made available
to them, they can resume their growth and
division
Osmotic pressure causes plasmolysis due to
the use of salts and sugars
High concentrations of these substances
create a hypertonic environment that causes
water to leave the microbial cell.
This process resemble desiccation

Objectives
Learn the terminology of microbial control
Study the physical methods of microbial control
Heat
Filtration
Low temperature
High pressure
Desiccation
Radiation

Radiation

Radiation has various effects on cells depending

on its wavelength intensity and duration
Ionizing radiation (X rays, gamma rays, electron
beams) has a wavelength shorter than that of
non-ionizing radiation, less than about 1 nm.
Therefore, it carries much more energy
Gamma rays penetrate deeply but may require
hours to sterilize large masses; high-energy
electron beams have much lower penetrating
power but usually require only a few seconds of
exposure.

 Ionizing radiation
Ionizes water to release OH
Damages DNA

Nonionizing radiation (UV, 260 nm) has a

wavelength longer than that of ionized
radiation, usually greater that about 1 nm
Damages DNA by causing bonds to form between
adjacent pyrimidine bases, usually thymines, in
DNA chains.
UV light can damage human eyes, and prolonged
exposure can cause burns and skin cancer

 Microwaves do not have much direct effect

on microorganisms, and bacteria can readily
be isolated from the interior of recently
operated microwave ovens.
Microwaves kill by heat; not especially
antimicrobial

The radiant energy spectrum.