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Chapter 7
Control of Microorganisms by Physical and Chemical Agents

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Figure 7.1
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Definition of Frequently Used Terms

sterilization
destruction or removal of all viable organisms

disinfection
killing, inhibition, or removal of pathogenic organisms disinfectants
agents, usually chemical, used for disinfection usually used on inanimate objects
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More definitions
sanitization
reduction of microbial population to levels deemed safe (based on public health standards)

antisepsis
prevention of infection of living tissue by microorganisms antiseptics
chemical agents that kill or inhibit growth of microorganisms when applied to tissue
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Antimicrobial agents
agents that kill microorganisms or inhibit their growth -cidal agents kill -static agents inhibit growth

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-cidal agents
-cide
suffix indicating that agent kills germicide
kills pathogens and many nonpathogens but not necessarily endospores

include bactericides, fungicides, algicides, and viricides

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-static agents
-static
suffix indicating that agent inhibits growth include bacteriostatic and fungistatic

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The Pattern of Microbial Death

microorganisms are not killed instantly population death usually occurs exponentially microorganisms were previously considered to be dead when they did not reproduce in conditions that normally supported their reproduction
however we now know that organisms can be in a viable but nonculturable (VBNC) condition
once they recover they may regain the ability to reproduce and cause infection
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Table 7.1
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Figure 7.2
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Conditions Influencing the Effectiveness of Antimicrobial Agent Activity

population size
larger populations take longer to kill than smaller populations

population composition
microorganisms differ markedly in their sensitivity to antimicrobial agents
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More conditions
concentration or intensity of an antimicrobial agent
usually higher concentrations or intensities kill more rapidly relationship is not linear

duration of exposure
longer exposure more organisms killed
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More Conditions
temperature higher temperatures usually increase amount of killing local environment many factors (e.g., pH, viscosity and concentration of organic matter) can profoundly impact effectiveness organisms in biofilms are physiologically altered and less susceptible to many antimicrobial agents
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The Use of Physical Methods in Control

heat low temperatures filtration radiation

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Moist Heat Sterilization

must be carried out above 100oC which requires saturated steam under pressure carried out using an autoclave effective against all types of microorganisms including spores degrades nucleic acids, denatures proteins, and disrupts membranes
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Table 7.2
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The Autoclave or Steam Sterilizer

Figure 7.3
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Pasteurization
controlled heating at temperatures well below boiling used for milk, beer and other beverages process does not sterilize but does kill pathogens present and slow spoilage by reducing the total load of organisms present
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Dry Heat Sterilization

less effective than moist heat sterilization, requiring higher temperatures and longer exposure times
items subjected to 160-170oC for 2 to 3 hours

oxidizes cell constituents and denatures proteins

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Dry Heat Incineration

bench top incinerators are used to sterilize inoculating loops used in microbiology laboratories
Figure 7.4
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Measuring Heat-Killing Efficiency

thermal death time (TDT)
shortest time needed to kill all microorganisms in a suspension at a specific temperature and under defined conditions

decimal reduction time (D or D value)

time required to kill 90% of microorganisms or spores in a sample at a specific temperature
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Z value
increase in temperature required to reduce D by 1/10

Figure 7.5
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Table 7.3
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Low Temperatures
freezing
stops microbial reproduction due to lack of liquid water some microorganisms killed by ice crystal disruption of cell membranes

refrigeration
slows microbial growth and reproduction
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Filtration
reduces microbial population or sterilizes solutions of heat-sensitive materials by removing microorganisms also used to reduce microbial populations in air
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Filtering liquids
depth filters
thick fibrous or granular filters that remove microorganisms by physical screening, entrapment, and/or adsorption

membrane filters
porous membranes with defined pore sizes that remove microorganisms primarily by physical screening
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Figure 7.6
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Figure 7.7
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Filtering air
surgical masks cotton plugs on culture vessels high-efficiency particulate air (HEPA) filters
used in laminar flow biological safety cabinets
Figure 7.8 (a)
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Figure 7.8 (b)

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Ultraviolet (UV) Radiation

limited to surface sterilization because it does not penetrate glass, dirt films, water, and other substances has been used for water treatment
Figure 7.9
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Ionizing Radiation
penetrates deep into objects destroys bacterial endospores; not always effective against viruses used for sterilization and pasteurization of antibiotics, hormones, sutures, plastic disposable supplies, and food
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Figure 7.10
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Table 7.4
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Table 7.5
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Chemical Control Agents Disinfectants and Antiseptics

Figure 7.11
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Phenolics

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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 disagreeable odor and can cause skin irritation
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Alcohols

bactericidal, fungicidal, but not sporicidal inactivate some viruses denature proteins and possibly dissolve membrane lipids
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Halogens
any of five elements: fluorine, chlorine, bromine, iodine, and astatine iodine and chlorine are important antimicrobial agents

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Halogens - Iodine
skin antiseptic oxidizes cell constituents and iodinates proteins at high concentrations may kill spores skin damage, staining, and allergies can be a problem iodophore
iodine complexed with organic carrier
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Halogens - Chlorine
oxidizes cell constituents important in disinfection of water supplies and swimming pools, used in dairy and food industries, effective household disinfectant destroys vegetative bacteria and fungi, but not spores can react with organic matter to form carcinogenic compounds
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Heavy Metals
e.g., ions of mercury, silver, arsenic, zinc, and copper effective but usually toxic combine with and inactivate proteins; may also precipitate proteins
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Quaternary Ammonium Compounds

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Quaternary Ammonium Compounds

detergents that have antimicrobial activity and are effective disinfectants
organic molecules with hydrophilic and hydrophobic ends act as wetting agents and emulsifiers

cationic detergents are effective disinfectants

kill most bacteria, but not Mycobacterium tuberculosis or endospores safe and easy to use, but inactivated by hard water and soap
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Aldehydes

highly reactive molecules sporicidal and can be used as chemical sterilants combine with and inactivate nucleic acids and proteins
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Effects of Glutaraldehyde

Figure 7.12
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Sterilizing Gases

used to sterilize heat-sensitive materials microbicidal and sporicidal combine with and inactivate proteins
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Figure 7.13
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Chemotherapeutic Agents
chemicals that can be used internally to kill or inhibit the growth of microbes within host cells their selective toxicity allows them to target the microbe without harming the host most are antibiotics, chemicals synthesized by microbes that are effective in controlling the growth of bacteria
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Evaluation of Antimicrobial Agent Effectiveness

complex process regulated by US federal agencies
Environmental Protection Agency Food and Drug Administration

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Phenol coefficient test

Table 7.6
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potency of a disinfectant is compared to that of phenol

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Other evaluation methods

use dilution test
determines rate at which selected bacteria are destroyed by various chemical agents

in-use testing
testing done using conditions that approximate normal use of disinfectant
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