Terminology of Microbial Control 2 Major Types of Microbial Control

I. Sterilization (all microbes)

sterilization II. Disinfection or antisepsis (vegetative cells and
 removal or destruction of all microbes, many viruses)
including viruses and bacterial a. Degerming
endospores, in or on an object b. Sanitization
 the eradication of harmful c. Pasteurization
microorganisms and viruses
microbial death
aseptic  permanent loss of reproductive ability
 environment or procedure that is free of under ideal environmental conditions
contamination of pathogens
microbial death rate
disinfection  calculating technique for evaluating the
 destruction of most microorganisms and efficacy of an antimicrobial agent which is
viruses on nonliving tissue (inanimate usually found to be constant over time for
objects) any particular microorganisms under a
 use of disinfectants that are more particular set of conditions
concentrated than antiseptics and can be
left on surface for longer periods of time
Action of Antimicrobial Agents
antisepsis
 reduction in the number of Alteration of Cell Walls and Membranes
microorganisms and viruses, particularly  a cell wall maintains cellular integrity by
potential pathogens, on living tissues counteracting the effects of osmosis when
 use of antiseptic the cell is in a hypotonic solution
 when damaged, effects of osmosis
degerming cause cells to burst
 removal of microbes from a surface by  cytoplasmic membrane acts as a bag that
scrubbing, such as when you wash your contains the cytoplasm and controls the
hands, or a nurse prepares an area of skin passage of chemicals into and out of the
for an injection cell
 chemicals such as soap or alcohol are  when damaged, cellular contents
commonly used leak out
 enveloped viruses have envelope that acts
sanitization as a membrane that is responsible for the
 process of disinfecting places and utensils attachment of the virus to its target cell
used by the public to reduce the number of  when damaged, viral replication is
pathogenic microbes to meet accepted prevented
public health standard  nonenveloped viruses can better tolerate
harsh conditions
pasteurization
 use of heat to kill pathogens and reduce Damage to Proteins and Nucleic Acids
the number of spoilage microorganisms in  proteins regulate cellular metabolism,
food or beverages function as enzymes in most metabolic
reactions, and form structural
-stasis or -static components in membranes and
 indicate that a chemical or physical agent cytoplasm
inhibits microbial metabolism and growth  denatured proteins cease to
but does not necessarily kill microbes function, bringing about cellular
death
-cide or -cidal  genes of a cell or virus are composed of
 indicate the agents that destroy or nucleic acids
permanently inactivate a particular  chemicals, radiation, and heat can
microbe alter or destroy nucleic acids
 disruption can produce fatal
mutations
 halt protein synthesis through
action on RNA
The Selection of Microbial Control  Effectiveness of germicides: (depends on
Methods the proficiency in inactivating or
destroying microorganisms that cannot
Ideal agents are: be sterilized with heat)
 inexpensive a. High-level germicides
 fast-acting o Kill all pathogens, including
 stable during storage bacterial endospores
 capable of controlling microbial growth o Used to sterilize invasive
while being harmless to humans, animals, instruments
and objects b. Intermediate-level germicides
Factors affecting the efficacy of Antimicrobial o Kill fungal spores, protozoan
Methods: cysts, viruses, and pathogenic
Site to be treated bacteria, but not bacterial
 harsh chemicals and extreme heat cannot endospores
be used on humans, animals, and fragile o used to disinfect instruments
objects that come in contact with
 when performing medical procedures, mucous membranes but are
medical personnel must choose a method noninvasive
and level of microbial control based on c. Low-level germicides
the site of the procedure o Eliminate vegetative bacteria,
fungi, protozoa, and some
Relative Susceptibility of Microorganisms viruses
o Used to disinfect items that
contact only the skin

Environmental Conditions
 Temperature and pH affect microbial
death rates and the efficacy of
antimicrobial methods
 Acidic conditions enhance the
antimicrobial effect of heat
 Clean objects before sterilization or
disinfection so that antimicrobial agents
 most resistant microbes: can thoroughly contact all the object’s
a. Bacterial endospores surfaces
o Endospores of Bacillus and
Clostridium are the most resilient Biosafety Levels:
forms of life Biosafety Level 1 (BSL-1)
o Can survive environmental  Handling microbes that do not cause
extremes of temperature, acidity, disease in healthy humans
and dryness and can withstand  Precautions are minimal and include
many chemical disinfectants handwashing with antibacterial soap and
b. Species of Mycobacteria disinfecting surfaces
o Cell walls of members of the
Biosafety Level 2 (BSL-2)
genus Mycobacterium such as
 Handling moderately hazardous agents
Mycobacterium tuberculosis,
 Procedures that may produce aerosols
contain a large amount of a waxy
are conducted within safety cabinets
lipid
o The wax allows the bacteria to Biosafety Level 3 (BSL-3)
survive drying and protects them  Stricter, requiring manipulations be done
from most water-based within safety cabinets containing high-
chemicals efficiency particulate air (HEPA) filters
o Must use strong disinfectants or  Entry through double sets of doors and
heat lower air pressure in the laboratory
c. Cysts of Protozoa Biosafety Level 4 (BSL-4)
o Protozoan cyst’s wall prevents entry  Handling dangerous or exotic microbes
of most disinfectants, protects that cause severe or fatal diseases in
against drying, and shields against humans
radiation and heat
  Lab space is isolated and personnel wear
protective suits
Physical Methods of Microbial Control  Autoclave
Heat-Related Methods o used to sterilize chemicals and
 Heat – older and more common means of objects that can tolerate moist
microbial control heat
 Effects of high temperature: o consists of pressure chamber,
 Denature proteins pipes to introduce and evacuate
 Interfere with the integrity of steam, valves to remove air and
cytoplasmic membranes and cell control pressure, and pressure
walls and temperature gauges to
 Disrupt the function and structure monitor the procedure
of nucleic acids c. Pasteurization
 Thermal death point – lowest  Developed by Louis Pasteur which
temperature that kills all cells in a broth is a method of heating beer and
in 10 minutes wine just enough to destroy the
 Thermal death time – time it takes to microorganisms that cause
completely sterilize a particular volume of spoilage without ruining the taste
liquid at a set temperature  Used to kill pathogens in milk, ice
cream, yogurt, and fruit juices
 Decimal reduction time (D) – time
 It is not sterilization because heat-
required to destroy 90% of the microbes
tolerant and heat-loving microbes
in a sample
survive
 Pasteurization of milk:
Moist Heat
o Batch method – 30 minutes at
 Commonly used to disinfect, sanitize,
63°C
sterilize, and pasteurize
o Flash pasteurization – high-
 Kills cells by denaturing proteins and
destroying cytoplasmic membranes temperature, short-time method
wherein milk flows through
 More effective in microbial control than
heated tubes that raise its
dry heat because water is a better
temperature to 72°C for only 15
conductor of heat than air
seconds
 Methods of moist heat:
o Ultra-high-temperature
a. Boiling
pasteurization – heats the milk
 Kills the vegetative cells of bacteria
to at least 135°C for only 1
and fungi, the trophozoites of
second
protozoa, and most viruses within
d. Ultra-High-Temperature Sterilization
10 minutes at sea level
 Involves flash heating milk or
 Boiling time is the critical factor –
other liquids to rid them of all
different elevations require
living microbes
different boiling time
 Involves passing the liquid through
 Bacterial endospores, protozoan
superheated steam at about 140°C
cysts, and some viruses can
for 1 to 3 seconds
survive boiling
 Treated liquids can be stored at
 Not recommended when true
room temperature
sterilization is required
 Effective for sanitizing restaurant
Dry Heat
tableware or disinfecting baby
bottles  Used for materials that cannot be
b. Autoclaving sterilized with moist heat
 True sterilization using heat  Hot air is an effective sterilizing agent
requires higher temperatures than because it denatures proteins and fosters
that of boiling water the oxidation of metabolic and structural
 Pressure is applied to boiling chemicals
water to prevent escape of heat  Requires higher temperatures for longer
 Temperature of 121°C, which times than moist heat because dry heat
requires the addition of 15 psi of penetrates more slowly
pressure above that of normal
pressure, destroys all microbes in Refrigeration and Freezing
a small volume in about 15  Decrease microbial metabolism, growth,
minutes and reproduction because chemical
 reactions occur more slowly at low  Energy without mass traveling in
temperatures because liquid water is not waves at the speed of light
available at subzero temperatures  The shorter the wavelength, the
 Refrigeration halts the growth of most more energy it carries
pathogens, which are predominantly  All types of radiation are described as
mesophiles either ionizing or nonionizing based on its
 Slow freezing, during which ice crystals effects on the chemicals within cells
have time to form and puncture cell
membranes, is more effective than quick Ionizing Radiation
freezing in inhibiting microbial  Wavelengths shorter than 1nm
metabolism  When they strike molecules, they have
sufficient energy to eject electrons from
Desiccation and Lyophilization atoms, creating ions
 Desiccation (drying)  Ions disrupt hydrogen bonding, oxidize
 Preserve foods such as fruits, peas, double covalent bonds, and create highly
beans, grain, nuts, and yeast reactive hydroxy radicals which in turn
 Inhibits microbial growth because denature other molecules, particularly
metabolism requires liquid water DNA, causing fatal mutations and cell
 Lyophilization (freeze-drying) death
 A technique that combines freezing  Nonionizing radiations:
and drying, to preserve microbes and a. Electron beams
other cells for many years  Produced by cathode ray
 Used for long-term preservation of machines
microbial cultures  Highly energetic and very
 Prevents formation of damaging ice effective in killing microbes in just
crystals few seconds
 Cannot sterilize thick objects or
Filtration objects coated with large amount
 Passage of a fluid through a sieve of organic matter
designed to trap particles and separate b. Gamma rays
them from the fluid  Emitted by some radioactive
 Viruses (filterable viruses) – pathogens elements
that are too small to be trapped in the  Penetrate much farther than
pores of filters electron beams but require hours
 Use filtration to estimate the number of to kill microbes
microbes in a fluid by counting the  Kills not only microbes but also
number deposited on the filter after larvae and eggs of insects
passing a given volume through the filter  Prevents both microbial spoilage
 Health care and laboratory workers and overripening
routinely use filtration to prevent c. X-rays
airborne contamination by microbes  Travel the fastest through matter
but have less energy than gamma
Osmotic Pressure rays
 Use of high concentrations of salt and  Require long time to kill microbes
sugar in foods to inhibit microbial growth  Not practical for microbial control
by osmotic pressure
 Removal of water inhibits cellular Nonionizing Radiation
metabolism because enzymes are fully  Wavelength greater than 1nm
functional only in aqueous environments  Does not have enough energy to force
 Fungi have greater ability than bacteria to electrons out of orbit but contains enough
survive hypertonic environments energy to excite electrons and cause them
to make new covalent bonds, which can
Radiation affect the 3-D structure of proteins and
 2 types of radiation: nucleic acids
a. Particulate radiation  Ultraviolet (UV) light
 Consists of high-speed subatomic  Has sufficient energy to be a practical
particles, such as protons, that antimicrobial agent
have been freed from their atoms  Causes pyrimidine dimers in DNA
b. Electromagnetic radiation  Does not penetrate well
 Suitable for disinfecting air,
transparent fluids, and surface of
objects
Chemical Methods of Microbial Control