Chapter 11

Physical and Chemical Agents for

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 Controlling Microorganisms
Primary targets of microbial control are microorganisms capable
of causing infection or spoilage that are constantly present in the
external environment and on the human body:
 bacterial vegetative cells and endospores
 fungal hyphae and spores, yeast
 protozoan trophozoites and cysts
 worms
 viruses
 prions

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 Relative Resistance of Microbes
Highest resistance
 bacterial endospores, prions
Moderate resistance
 viruses - naked viruses: hepatitis B virus, poliovirus
 Mycobacterium tuberculosis, Staphylococcus aureus, Pseudomonas sp.
 protozoan cysts
 some fungal sexual spores (zygospores)
Least resistance
 most bacterial vegetative cells
 fungal spores and hyphae, yeasts
 enveloped viruses
 protozoan trophozoites

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 Terminology and Methods of Control
Through the years, a growing terminology has emerged for
describing and defining measures that control microbes
These measures include:
 sterilization
 disinfection
 asepsis and antisepsis
 sanitization
 degermation

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 Sterilization
Sterilization
is a process that destroys or removes all viable
microorganisms, including viruses and endospores
Any material that has been subjected to this process is said to be
sterile
An object is either “sterile” or “not sterile”
Control methods that sterilize are generally reserved for
inanimate objects
Sterilized products are essential to human well-being
Chemicals called sterilants are also used as sterilizing agents

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 Microbicidal Agents
-cide means to kill
Bactericide - destroys bacteria (except endospores)
Fungicide - kills fungal spores, hyphae, and yeasts
Virucide - inactivate viruses especially on living tissue
Sporicide - destroys bacterial endospores

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 Microbistasis
Microbistasisis a condition in which microbes are temporarily
prevented from multiplying but are not killed outright
Bacteriostatic agents prevent bacterial growth on tissues or on
objects in the environment
Fungistatic chemicals inhibit fungal growth
Antiseptics
and drugs used to control microorganisms in the body
have microbistatic effects

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 Disinfection
Disinfection is the use of a physical process or a chemical agent
(disinfectant) to destroy vegetative pathogens but not bacterial
endospores
Normally used only on inanimate objects
The concentrations required to be effective can be toxic to human
and other animal tissue
Disinfection
processes also remove the harmful products of
microorganisms (toxins) from materials

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 Asepsis and Antisepsis
Asepsis refers to any practice that prevents the entry of infectious
agents into sterile tissues and thus prevent infection
Antisepsis is the application of chemical agents (antiseptics)
directly to exposed body surfaces, wounds, and surgical incisions
to destroy or inhibit vegetative pathogens
Commonly practiced in health care

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 Sanitization
Sanitizationis referred to as any cleansing technique that
mechanically removes debris, microorganisms, and toxins
Sanitization reduces the potential for infection and spoilage
Soaps and detergents are the most commonly employed sanitizers
Sanitization is in general far less expensive than sterilization
Applies to inanimate objects and surfaces

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 Degermation
Degermation refers to the reduction in microbial load on a living
tissue through mechanical means
Usually involves scrubbing the skin or immersing it in chemicals,
or both
Microorganisms are removed both mechanically and, indirectly
by emulsifying the oils that coat the skin and provide a habitat for
their growth

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 Microbial death
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

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 Practical Concerns in Microbial Control
Selection of method of control depends on circumstances:
 does the application require sterilization or is disinfection adequate?
 is the item to be reused or permanently discarded?
 can the item withstand heat, pressure, radiation, or chemicals?
 is the control method suitable for a given application?
 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:
 the cell wall – cell wall becomes fragile and cell lyses; some
antimicrobial drugs, detergents, and alcohol
 the cell membrane - loses integrity; detergent surfactants
 cellular synthetic processes (DNA, RNA) – prevention of replication,
transcription; some antimicrobial drugs, radiation, formaldehyde,
ethylene oxide
 proteins – interfere at ribosomes to prevent translation, disrupt or
denature proteins; alcohols, phenols, acids, heat

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 Methods of Physical Control
Heat – moist and dry
Cold temperatures
Desiccation
Radiation
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
Bacterialendospores are the most resistant – usually require
temperatures above boiling
Vegetative states of bacteria and fungi are the least resistant

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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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 Common Methods of Moist Heat Control
The four ways that moist heat is employed to control microbes
are:
 steam under pressure
 nonpressurized steam
 boiling water
 pasteurization

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 Steam Under Pressure
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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 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
Boilingat 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
63oC – 66oC for 30 minutes (batch method)
71.6oC 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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 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
Noteffective 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
 used to sterilize medical supplies and food products
Nonionizing radiation – little penetrating power – must be
directly exposed
UV light creates thymine dimers, which interfere with replication

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 Sterilization by 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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 Chemical Agents in Microbial Control
Antimicrobial chemicals occur in the liquid, gaseous, or even
solid state
They serve as disinfectants, antiseptics, sterilants, degermers, and
preservatives (chemicals that inhibit the deterioration of
substances)

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 Desirable Qualities in a Germicide
rapid action in low concentrations
solubility in water or alcohol and long-term stability
broad spectrum, non toxic to human and animal tissues
penetration of inanimate surfaces to sustain a cumulative or
persistent action
resistant to becoming inactivated by organic matter
noncorrosive or nonstaining properties
sanitizing and deodorizing properties
affordability and ready availability

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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 microorganisms being treated
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
Halogens
Phenolics
Chlorhexidine
Alcohols
Hydrogen peroxide
Detergents and soaps
Heavy metals
Aldehydes
Gases
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)
 denature proteins
 intermediate level
 milder medical & 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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 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
Weak (3%) to strong (25%)
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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 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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 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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 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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 Gases and Aerosols
Ethylene oxide, propylene oxide
Strong alkylating agents
High level
Sterilize and disinfect plastics and prepackaged devices, foods

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