Foundations in

Microbiology
Seventh Edition

Talaro
Chapter 11
Physical and Chemical
Agents for Microbial
Control
 11.1 Controlling Microorganisms
• Physical, chemical, and mechanical methods to
destroy or reduce undesirable microbes in a given
area (decontamination)
• Primary targets are microorganisms capable of
causing infection or spoilage:
– Vegetative bacterial cells and endospores
– Fungal hyphae and spores, yeast
– Protozoan trophozoites and cysts
– Worms
– Viruses
– Prions

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Figure 11.1

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 Relative Resistance of Microbes
• Highest resistance
– Prions, bacterial endospores
• Moderate resistance
– Pseudomonas sp.
– Mycobacterium tuberculosis
– Staphylococcus aureus
– Protozoan cysts
• Least resistance
– Most bacterial vegetative cells
– Fungal spores and hyphae, yeast
– Enveloped viruses
– Protozoan trophozoites 4
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 Terminology and Methods of Control
• Sterilization – a process that destroys all viable
microbes, including viruses and endospores
• Disinfection – a process to destroy vegetative
pathogens, not endospores; inanimate objects
• Antiseptic – disinfectants applied directly to exposed
body surfaces
• Sanitization – any cleansing technique that
mechanically removes microbes
• Degermation – reduces the number of microbes
through mechanical means
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 Microbial Death
• Hard to detect, microbes often reveal
no conspicuous vital signs to begin with
• Permanent loss of reproductive capability,
even under optimum growth conditions

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 Factors That Affect Death Rate
The effectiveness of a particular agent
is governed by several factors:
• Number of microbes
• Nature of microbes in the
population
• Temperature and pH of environment
• Concentration or dosage of agent
• Mode of action of the agent
• Presence of solvents, organic matter, or
inhibitors 9
Figure 11.2

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Practical Concerns in Microbial Control
Selection of method of control depends on circumstances:
• Does the application require sterilization?
• Is the item to be reused?
• Can the item withstand heat, pressure, radiation,
or chemicals?
• Is the method suitable?
• Will the agent penetrate to the necessary extent?
• Is the method cost- and labor-efficient and is it
safe?
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 Antimicrobial Agents’ Modes of Action
Cellular targets of physical and chemical agents:
1. The cell wall – cell wall becomes fragile and cell
lyses; some antimicrobial drugs, detergents, and
alcohol
2. The cell membrane – loses integrity; detergent
surfactants
3. Protein and nucleic acid synthesis – prevention of
replication, transcription, translation, peptide
bond formation, protein synthesis;
chloramphenicol, ultraviolet radiation,
formaldehyde
4. Proteins – disrupt or denature proteins; alcohols,
phenols, acids, heat
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Figure 11.3

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Figure 11.4

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11.2 Methods of Physical Control

1. Heat – moist and dry

2. Cold temperatures
3. Desiccation
4. Radiation
5. Filtration

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 Mode of Action and Relative
Effectiveness of Heat
• Moist heat – lower temperatures and shorter
exposure time; coagulation and
denaturation of proteins
• Dry heat – moderate to high temperatures;
dehydration, alters protein structure;
incineration

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 Heat Resistance and Thermal Death
• Bacterial endospores most resistant –
usually require temperatures above
boiling

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 Thermal Death Measurements
• Thermal death time (TDT) – shortest
length of time required to kill all test
microbes at a specified temperature
• Thermal death point (TDP) – lowest
temperature required to kill all microbes in
a sample in 10 minutes

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 Moist Heat Methods
• Steam under pressure – sterilization
• Autoclave 15 psi/121oC/10-40min
• Steam must reach surface of item
being sterilized
• Item must not be heat or moisture
sensitive
• Mode of action – denaturation of
proteins, destruction of membranes and
DNA
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Figure 11.5

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 Nonpressurized Steam
• Tyndallization – intermittent sterilization for
substances that cannot withstand autoclaving
• Items exposed to free-flowing steam for 30–60
minutes, incubated for 23–24 hours and then
subjected to steam again
• Repeat cycle for 3 days
• Used for some canned foods and laboratory
media
• Disinfectant

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 Boiling Water
• Boiling at 100oC for 30 minutes to destroy
non-spore-forming pathogens
• Disinfection

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 Pasteurization
• Pasteurization – heat is applied to kill
potential agents of infection and spoilage
without destroying the food flavor or value
• 63 C–66 C for 30 minutes (batch method)
• 71.6 C for 15 seconds (flash method)
• Not sterilization – kills non-spore-forming
pathogens and lowers overall microbe count;
does not kill endospores or many
nonpathogenic microbes
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 Dry Heat
Dry heat using higher temperatures
than moist heat
• Incineration – flame or electric
heating coil
– Ignites and reduces microbes and other
substances
• Dry ovens – 150–180oC – coagulate
proteins

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Figure 11.6

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 Cold
• Microbiostatic – slows the growth of
microbes
• Refrigeration 0–15oC and freezing
<0oC
• Used to preserve food, media, and
cultures

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 Desiccation
• Gradual removal of water from cells,
leads to metabolic inhibition
• Not effective microbial control – many cells
retain ability to grow when water is
reintroduced
• Lyophilization – freeze drying;
preservation

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 Radiation
• Ionizing radiation – deep penetrating power
that has sufficient energy to cause electrons to
leave their orbit, breaks DNA
– Gamma rays, X-rays, cathode rays
– Cold (low temperature) sterilization
– Used to sterilize medical supplies and food
products

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Figure 11.6

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Figure 11.8

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 Radiation
• Nonionizing radiation – little penetrating
power – must be directly exposed
• UV light creates pyrimidine dimers,
which interfere with replication

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Figure 11.9

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 Filtration
• Physical removal of microbes by passing
a gas or liquid through filter
• Used to sterilize heat sensitive liquids and
air in hospital isolation units and industrial
clean rooms

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Figure 11.1

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 11.3 Chemical Agents in
Microbial Control
• Disinfectants, antiseptics, sterilants, degermers,
and preservatives
• Some desirable qualities of chemicals:
– Rapid action in low concentration
– Solubility in water or alcohol, stable
– Broad spectrum, low toxicity
– Penetrating
– Noncorrosive and nonstaining
– Affordable and readily available

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Levels of Chemical Decontamination
• High-level germicides – kill endospores; may be
sterilants
– Devices that are not heat sterilizable and intended to be
used in sterile environments (body tissue)
• Intermediate-level – kill fungal spores (not endospores),
tubercle bacillus, and viruses
– Used to disinfect devices that will come in contact
with mucous membranes but are not invasive
• Low-level – eliminate only vegetative bacteria,
vegetative fungal cells, and some viruses
– Clean surfaces that touch skin but not mucous
membranes
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 Factors that Affect Germicidal
Activity of Chemicals
• Nature of the material being treated
• Degree of contamination
• Time of exposure
• Strength and chemical action of
the germicide

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 Germicidal Categories
1. Halogens
2. Phenolics
3. Chlorhexidine
4. Alcohols
5. Hydrogen peroxide
6. Detergents & soaps
7. Heavy metals
8. Aldehydes
9. Gases
10. Dyes

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 Halogens
• Chlorine – Cl2, hypochlorites (chlorine bleach),
chloramines
– Denaturate proteins by disrupting disulfide bonds
– Intermediate level
– Unstable in sunlight, inactivated by organic matter
– Water, sewage, wastewater, inanimate objects

• Iodine - I2, iodophors (betadine)

– Interferes with disulfide bonds of proteins
– Intermediate level
– Milder medical and dental degerming agents, disinfectants,
ointments

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 Phenolics
• Disrupt cell walls and membranes
and precipitate proteins
• Low to intermediate level – bactericidal,
fungicidal, virucidal, not sporicidal
– Lysol
– Triclosan – antibacterial additive to
soaps

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Figure 11.2

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 Chlorhexidine
• A surfactant and protein denaturant
with broad microbicidal properties
• Low to intermediate level
• Hibiclens, Hibitane
• Used as skin degerming agents for
preoperative scrubs, skin cleaning,
and burns

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 Alcohols
• Ethyl, isopropyl in solutions of 50-95%
• Act as surfactants dissolving membrane
lipids and coagulating proteins of
vegetative bacterial cells and fungi
• Intermediate level

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 Hydrogen Peroxide
• Produce highly reactive hydroxyl-free
radicals that damage protein and DNA
while also decomposing to O2 gas – toxic to
anaerobes
• Antiseptic at low concentrations;
strong solutions are sporicidal

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Figure 11.13

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 Aldehydes
• Glutaraldehyde and formaldehyde kill by alkylating
protein and DNA
• Glutaraldehyde in 2% solution (Cidex) used as
sterilant for heat sensitive instruments
• High level
• Formaldehyde – disinfectant, preservative, toxicity
limits use
– Formalin – 37% aqueous solution
• Intermediate to high level

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Figure 11.14

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 Gases and Aerosols
• Ethylene oxide, propylene oxide
• Strong alkylating agents
• High level
• Sterilize and disinfect plastics and prepackaged
devices, foods

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Figure 11.15

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 Detergents and Soaps
• Quaternary ammonia compounds (quats) act as
surfactants that alter membrane permeability
of some bacteria and fungi
• Very low level
• Soaps – mechanically remove soil and
grease containing microbes

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Figure 11.16

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Figure 11.17

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 Heavy Metals
• Solutions of silver and mercury kill
vegetative cells in low concentrations
by inactivating proteins
• Oligodynamic action
• Low level
• Merthiolate, silver nitrate, silver

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Figure 11.18

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 Dyes as Antimicrobial Agents
• Aniline dyes are very active against
gram- positive species of bacteria and
various fungi
• Sometimes used for antisepsis and wound
treatment
• Low level, narrow spectrum of activity

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 Acids and Alkalis
• Low level of activity
– Organic acids prevent spore germination and
bacterial and fungal growth
– Acetic acid inhibits bacterial growth
– Propionic acid retards molds
– Lactic acid prevents anaerobic bacterial
growth
– Benzoic and sorbic acid inhibit yeast

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