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Chapter 08
Control of
Microorganisms in
the Environment
Prescott’s Microbiology
Twelfth edition
Joanne Willey, Kathleen
Sandman, Dorothy Wood

© 2023 McGraw Hill, LLC. All rights reserved. Authorized only for instructor use in the classroom.
No reproduction or further distribution permitted without the prior written consent of McGraw Hill, LLC.
 8.1 Microbial Growth and Replication: Targets for
Control

a. Compare and contrast actions of disinfection, antisepsis,

chemotherapy, and sterilization.
b. Distinguish between cidal (lethal) and static (inhibitory)
agents.
c. Calculate the decimal reduction time (D value).
d. Correlate antisepsis, sanitization, disinfection, and
sterilization with agent effectiveness.

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 Common Microbial Control Methods

Biocide–Antimicrobial agents that control microorganisms.

• Physical, chemical, mechanical or biological.

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 Definition of Frequently Used Terms

Sterilization—process by which all living cells, spores, and acellular

entities are destroyed or removed from an object.
• Sterilant is the chemical agent.
Disinfection—killing, inhibition, or removal of disease-causing
microorganisms.
• Disinfectants—agents, usually chemical, used for disinfection,
usually used on inanimate objects.
• Does not always sterilize as spores or a few organisms can remain.
Sanitization—reduction of microbial population to levels deemed safe by
public health standards.
Antisepsis—destruction of microbes on living tissue.
• Antiseptics—chemical agents applied to tissue to kill or inhibit growth
of pathogen.
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 Antimicrobial Agents

Chemotherapy—generic term that describes application of

chemicals to kill microorganisms.
Cidal agents kill (-cide suffix indicates agent that kills).
• May be especially effective against a specific group.
• Include bactericides, fungicides, and viricides.

Static agents inhibit growth (-static suffix indicates growth

inhibiting agent).
• May be especially effective against a specific group.
• Include bacteriostatic and fungistatic.

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 Resistance to Biocides

Resistance to antimicrobial biocides has been increasing

similar to the rate of resistance to antibiotics.
Mechanisms of resistance similar to antibiotic resistance.
• Efflux pumps
• Alter membrane permeability
• Modify the target
• Specific resistance genes

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 The Pattern of Microbial Death Mirrors the Pattern of
Microbial Growth

Microorganisms are not killed instantly when exposed to

lethal agent.
Population death usually occurs exponentially.
• Reduced by the same fraction at constant intervals.

Decimal reduction time—time required to kill 90% of

microorganisms.
• D value is time required to drop by 10-fold.
• Z value is the temperature change that decreases the
microbial population by 90%.

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 D and Z Values

Minute Microbial Number Microorganisms Killed Viable Microorganisms Log10 of

at Start of Minute in 1 Minute (90% of total) at End of 1 Minute Survivors
1 106 9  105 105 5
2 105 9  104 104 4
3 104 9  103 103 3
4 103 9  102 102 2
5 102 9  101 10 1
6 101 9 1 0
7 1 0.9 0.1 −1

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 Impact of Biocide Exposure

Three possible population reduction curves from different

biocides.

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 8.2 Microbes Can Be Controlled by Physical Means

a. Explain the mechanism by which filtration removes

microorganisms.
b. Differentiate between methods of filter sterilizing liquids
and gases.
c. Describe the application of heat and radiation to control
microorganisms.
d. Explain the mechanisms by which heat and radiation
control microbes.

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 Filtration

Reduces microbial population in heat-

sensitive materials by removing
microorganisms.
Also used to reduce microbial
populations in air.
Depth filter—fibrous materials that
have been bonded into a think layer
filled with narrow, twisting channels.
• Solution with microbes is sucked
through the layer under vacuum
and microbes adsorb to the surface
of the filter material.

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 Filtering Liquids

Membrane filters
• Porous membranes with defined
pore sizes that remove
microorganisms.
• Often used after a depth filter.

(c ) Callista Images Cultura/Newscom

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 Filtering Air

Surgical masks
• N95 disposable mask
• Exclude 95% of particles.
(a) James Gathany/CDC
High-efficiency particulate air
(HEPA) filters
• Used in laminar flow
biological safety cabinets.
• Exclude 99.97% of particles.
• During the COVID-19
pandemic, airlines showed
this filter to be effective.

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 Moist Heat

Destroys viruses, fungi, and bacteria by degrading nucleic

acids, denaturing proteins, and disrupting cell membranes.
Boiling will not destroy endospores and does not sterilize.
• Can be used to disinfect drinking water.
Average conditions for moist heat inactivation:
Entity Vegetative Cells Spores
Yeasts 5 minutes at 50 to 60°C 5 minutes at 70 to 80°C

Molds 30 minutes at 62°C 30 minutes at 80°C

Bacteria 10 minutes at 60 to 70°C 2 to over 800 minutes at 100°C 0.5
to 12 minutes at 121°C
Viruses 30 minutes at 60°C
Prions 90 minutes at 134°C

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 Steam Sterilization

Autoclave—device used in steam

sterilization.
Carried out above 100oC which
requires saturated steam under
pressure.
(a) BERANGER/BSIP SA/Alamy Stock Photo

Effective against all types of

microorganisms, including spores.
Quality control
• Geobacillus stearothermophilus
strips or color changing tape.

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 Pasteurization

• Controlled heating at temperatures well below boiling.

• Used for milk, wine, and other beverages.
• Process does not sterilize, but does kill pathogens and
slows spoilage by reducing the total load of organisms
present.

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 Tyndallization

For materials that cannot withstand the high temperature of

the autoclave.
Intermittent sterilization
• 30 to 60 minutes of steam exposure with 23 to 24 hours
between incubations.
• 2 or 3 times this is repeated.

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 Dry Heat Sterilization

Less effective than moist heat sterilization, requiring higher

temperatures and longer exposure times.
• Items subjected to 160 to 170oC for 2 to 3 hours.

Oxidizes cell constituents and denatures proteins which

causes microbial death.
Not suitable for heat-sensitive materials (that is, plastic)
Advantage
• Does not corrode glassware and metal instruments.

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 Ultraviolet (UV) Radiation

• Wavelength of 260 nm is most

lethal.
• Causes thymine dimers
preventing replication and
transcription.
• UV limited to surface
sterilization because it does
not penetrate glass, dirt films,
water, and other substances.
Sundry Photography/Shutterstock

• Has been used for water

treatment.

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 Cold Plasma

Fourth state of matter.

Made of ionized gases where the atoms and/or molecules

are stripped of outer-shell electrons.

Emits UV radiation and reactive oxygen and/or nitrogen

species.
• Damages nucleic acids
• Oxidizes nucleic acids, proteins, and lipids.

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 Ionizing Radiation

• Gamma radiation penetrates deep into objects.

• Dislodges electrons from atoms or molecules, producing
chemically reactive free radicals.
• Used for sterilization and pasteurization of antibiotics,
hormones, sutures, plastic disposable supplies, and food.

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 8.3 Microorganisms Are Controlled with Chemical
Agents

a. Describe the use and mechanism of action for phenolics,

alcohols, halogens, heavy metals, quaternary ammonium
compounds, aldehydes, and oxides to control
microorganisms.
b. Design novel antimicrobial control applications using
phenolics, alcohols, halogens, metals, quaternary
ammonium compounds, aldehydes, and oxides.

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 Chemical Control Agents and Their Requirements

Disinfection
• Ideally the biocide is effective against wide variety of
infectious agents at low concentrations and in the
presence of organic matter.
• Balance between effectiveness and low toxicity.

Antisepsis
• Reduce number of pathogens on human tissues to
prevent infection.

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 Phenolics

• Commonly used as laboratory and hospital disinfectants.

• Act by denaturing proteins and disrupting cell membranes.
• Tuberculocidal, effective in presence of organic material, and
long lasting.
• Lysol®, a commercial disinfectant, is a mixture of phenolics.
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 Alcohols

• Among the most widely used disinfectants, antiseptics,

and sanitizers.
• Two most common are ethanol and isopropanol.
• Bactericidal, fungicidal, but not sporicidal.
• Inactivate some viruses.
• Act by denaturing proteins and dissolving membrane
lipids.
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 Halogens—Iodine

Skin antiseptic.
Acts by oxidizing cell constituents and iodinating proteins.
At high concentrations may kill endospores.
May cause skin damage, staining, and allergies can be a
problem.
Iodophor
• Iodine complexed with organic carrier.
• Released slowly to minimize skin burns.

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 Halogens—Chlorine

• Acts by oxidizing cellular materials and destroys

vegetative bacteria and fungi.

• Important in disinfection of water supplies, swimming

pools, and used in dairy and food industries.

• Chlorine is a household disinfectant as well because it is

inexpensive and effective.

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 Metals

Ions of mercury, silver, arsenic, zinc, and copper were used

for many years as germicides.
• Silver and copper only used now.

Silver sulfadiazine is used topically on burns.

Copper sulfate is an effective algicide in lakes and swimming

pools.

Act by inactivating proteins, often via sulfhydryl groups.

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 Quaternary Ammonium Compounds

Detergents that have broad spectrum antimicrobial activity and are

effective disinfectants.
• Detergents—amphipathic organic cleansing agents.
Cationic detergents are effective disinfectants.
• Kill most bacteria, but not M. tuberculosis or endospores.
• Stable and nontoxic, but inactivated by hard water and soap.
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 Aldehydes

• Commonly used agents are formaldehyde and

glutaraldehyde.
• Highly reactive molecules that act by inactivating nucleic
acids and proteins.
• Sporicidal and can be used as chemical sterilants.

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 Effects of Glutaraldehyde

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 Sterilizing Gases

• Used to sterilize heat-

sensitive materials.
• Microbicidal and
sporicidal.
• Ethylene oxide
sterilization is carried out (a) Anderson Products, www.anpro.com

in equipment resembling
an autoclave.
• Vaporized hydrogen
peroxide can also be
used.

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 8.4 Antimicrobial Agents Must Be Evaluated for
Effectiveness

a. Predict the effects of microbial population size and

composition, temperature, exposure time, and local
environmental conditions on antimicrobial agent
effectiveness.
b. Describe the processes used to measure microbial killing
rates, dilution testing, and in-use testing of antimicrobial
agents.

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 Evaluation of Antimicrobial Agent Effectiveness

Complex process regulated by two U.S. federal agencies:

• Environmental Protection Agency regulates disinfectants.
• Food and Drug Administration regulates agents used on
humans and animals.
Establish guidelines under which these agents are used and
agent effectiveness is measured.

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 Conditions Influencing the Effectiveness of
Antimicrobial Agent Activity 1

Population size
• Larger populations take longer to kill than smaller
populations.
Population composition
• Microorganisms differ markedly in their sensitivity to
antimicrobial agents (endospores more resistant).
Concentration or intensity of an antimicrobial agent
• Usually higher concentrations kill more rapidly.
• Not a linear relationship, small changes can have large effect.

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 Conditions Influencing the Effectiveness of
Antimicrobial Agent Activity 2

Contact time
• The longer the exposure, the more organisms killed.

Temperature
• Higher temperatures enhance chemical activity.

Local environment
• Population to be controlled is not isolated but surrounded
by environmental factors that can either prevent or aid in
destruction.
• pH, viscosity, and concentration of organic matter.
• Organisms in biofilms are less susceptible to many antimicrobial
agents.

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 Efficiency Evaluation of Chemical Agents

Phenol coefficient test

• Potency of a disinfectant is compared to that of phenol.
• Useful for initial screening but may be misleading.

Use dilution test

• Determines rate at which selected bacteria are destroyed
by various chemical agents.

Normal in-use testing

• Testing done using conditions that approximate normal
use of disinfectant.

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 8.5 Microorganisms Can Be Controlled by Biological
Methods

a. Propose predation, competition, and other methods for

biological control of microorganisms.

b. Suggest alternative decontamination and medical

therapies using viruses of bacteria, fungi, and protozoa.

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 Biological Control of Microorganisms

Emerging field showing great promise.

Natural control mechanisms:
• Predation by Bdellovibrio.
• Bacteriophages
• A spray format recently approved by the F DA can be used on food
products.
• Enzybiotics
• Proteins purified from bacteriophage that cause host cell lysis.
• Toxin-mediated killing using bacteriocins.

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 Take Home Message

• Controlling microbial growth can be achieved by many

different methods—some physical, some chemical.
• Understanding what microbes need to grow and thrive is
important for then understanding how to kill them off or
control their growth.
• While many of these methods are useful, each has pros
and cons—and each may not be suitable in particular
circumstances.
• Always be thinking about what the appropriate and most
useful way to control microbe growth in a given
environment or situation might be using the information
found in this chapter.

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© 2023 McGraw Hill, LLC. All rights reserved. Authorized only for instructor use in the classroom.
No reproduction or further distribution permitted without the prior written consent of McGraw Hill, LLC.