STERILIZATION AND
DISINFECTION
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Learning Objectives:
At the end of this chapter, the student will be able to:
1. Define sterilization and disinfection.
2. Identify the various ways of sterilization and disinfection
techniques.
3. Explain why materials are sterilized in medical laboratory.
4. Describe how to disinfect and decontaminate laboratory
waste.
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Outline
STERILIZATION AND DISINFECTION
Definitions
Methods of sterilization.
Factors affecting sterilization and disinfection
Controlling of sterilization
Disinfection and decontamination of laboratory wastes
Disinfection
Decontamination using chemical disinfectants
Decontamination of laboratory space
Incineration
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Definitions
Sterilization: Destruction of all forms of microbial life
including spores.
Disinfection: Disinfection of microbes that cause disease;
may not be effective in killing spores
Antiseptics: Are chemicals applied in living tissue which are
able to destruct or inhibit the growth of microorganisms
there by limiting or preventing the harmful effect of infection.
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Methods of sterilization
Sterilization and disinfecting agents are divides into two groups.
These are:
Chemical means’s of sterilization and disinfection.
Physical means’s of sterilization
I. Chemical means’s of sterilization & disinfection
These chemical agents destroy any type of microbes with
out showing any form of selectivity unlike antibiotics.
Classification of chemical mean’s of sterilization and
disinfection
1.Chemical agents that damage the cell membrane
Surface Active Agents
Phenols
Organic solvents ( Alcohol e.g. Ethyl alcohol,
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Methods……
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b)Oxidizing agents
Converts functional-SH group into non-functional-S-S
group.
Halogens e.g. Chlorine, iodine
Chlorine: inactivated by organic materials.
c). alkaylating agents
E.g. Formaldehyde, Glutaraldehdyde ( 10X more effective than
formaldehyde)
d). Anti-septic agents
Disinfectants that are applied on an animate bodies
Never be toxic to cells,
never be corrosive,
should never change nature of skin
E.g. Savlon, 70% alcohol, Tincture of Iodine
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2. Physical methods
1. Heat
It is the most reliable and universally applicable method
Types
A. Dry heat
B. Moist heat
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A. Dry heat
It is less efficient and requires high temperature and long period heating
than moist heat.
a. Incineration: It is an efficient method of sterilization and disposal of
contaminated needles, syringes and cover slips at high temperature.
b. Red heat: Inoculating wires, loops and points of forceps are sterilized by
holding them in the flame of a Bunsen burner until they are red hot.
c. Flaming: Scalpels and neck of flasks, bottles and tubes are exposed for
a few seconds, but it is of uncertain efficacy.
d. Hot Air Sterilizer (Oven): it is essential that hot air should circulate
between the objects being sterilized and these must be loosely packed
and adequate air space to ensure optimum heat transfer. It is done by
applying 1600c for 1 hour.
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B. Moist heat:
It
is preferred to dry heat due to more rapid killing.
Moist heat can be used by the following methods.
1. Boiling: It is not reliable method of sterilization.
It is done by applying 1000c for 30 minutes.
Used for sterilizing catheters, dressing
2. Tyndallization: Intermittent steaming
Steaming of the material is done at 1000c for 30 minutes on
three consecutive days.
The principle is that spores which survived the heating
process would germinate before the next thermal
exposure and then would be killed.
It is used for sterilizing heat sensitive culture media
containing materials such as carbohydrates, egg or
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3. Pasteurization:
It is the process of application of heat at temperature of 620c
for 30 minutes or 720c for 15 seconds followed by rapid
cooling to discourage bacterial growth.
Uses:
Pasteurization of milk
Preparation of bacterial vaccines.
4. Autoclave: Steam under pressure
It is based on the principle that when microorganism is
boiled at increased pressure, hot saturated steam will be
formed which penetrates and gives up its latent heat when it
condenses on objects.
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Autoclaving …
Uses: Sterilize solid and fluid culture media, gowns,
medical and surgical equipment.
Time-Temperature-Pressure Level Relationship in
moist heat sterilization (Autoclaving)
Temperature Time Pressure level
1210c 15 minutes 15 Ib/inch2
1260c 10 minutes 20 Ib/inch2
1340c 3 minutes 30 Ib/inch2
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Other methods
5.freezing ( lyophilization)
6. Filtration
7. Radiation
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Factors affecting sterilization and disinfection
The efficacy of the above on the of methods following factors.
Concentration of the agent
There is a relationship between the concentration of the agent and
the time required to kill a given fraction of the microbial population.
Time of exposure
Microbes are killed with a reasonable length of time with chemical
agents.
PH of the medium; where action determines degree of
ionization of the chemical and bacterial surface charge.
Temperature
Bactericidal potency of the chemical agent increases with an
increase in temperature.
An increase in 100c doubles the bacterial death rate.
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Factors…..
Natureof the organism
Species of the bacteria
Growth phase of bacteria in culture
Presence of capsule, spore and other special structures
Number of bacteria in test system
Presence of extraneous materials
Organic materials like serum, blood or pus makes chemicals inert that
are highly active in their absence.
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Methods of Controlling Sterilization
1. Following scientific procedures strictly.
2. Using color indicator:
e.g. Heat-sensitive autoclave tape fixed to the outside of
each pack.
Color change of autoclave tape from blue to brown-black indicates
complete sterilization.
3. Biological indicator:
Use of paper strips impregnated with spores of Bacillus
thermophilus.
Put the paper strip in the culture medium after autoclaving and
observe for germinating bacteria to check for growth.
In a complete sterilization there should not be bacterial growth.
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