DISINFECTION , AND STERILIZATION
Learning objectives
Up on completion of this chapter, the student will be
able to:
– Define disinfection, antiseptic and sterilization.
– Describe means of sterilization and disinfection.
– Discuss the principles of sterilization and
disinfection.
Sterilization & disinfection
Definition of terms:
Sterilization:
Killing or removing all forms of microbial life in a
material or an object.
involves the removal of both vegetative or endospores.
Disinfection:
Reducing the number of pathogenic microorganisms
capable of giving rise to infection.
Usually involves the removal of Vegetative form of
pathogens.
May not be effective in killing spores.
3
Definition of terms(cont…)
• Antisepsis: destruction or inhibition of micro-
organisms in living tissue
Abate A. 4
Methods of sterilization & disinfection
Abate A. 5
I. Physical means of sterilization and disinfection
– Physical means of sterilization and disinfection
include Heat, Filtration and Radiation
I. Heat: is the most reliable and universally
applicable method of sterilization.
Mechanism of Action:
• Dry heat-oxidative damage and denatures
protein.
• Moist heat-coagulation and denaturation of
proteins
Abate A. 6
Physical means…
1.1. Dry Heat
Dry heat sterilization requires higher temperature and
often takes longer than moist heat sterilization.
The longer time and higher temperature require in a
dry heat sterilization is because
heat in water can ready transfer to cold object
than heat in air.
Abate A. 7
Dry heat (cont’d)
A. Incineration:
Effective way to sterilize disposable items (paper cups,
dressings) and biological waste.
B. Flaming:
It is a technique of passing an object over a flame with
out allowing it to become red hot.
Used to disinfect glass slides, mouth of culture test
tubes etc.
C. Red hot:
Sterilization of an object by holding them in a flame
till they become red hot
It is used for sterilizing Abate
needles and inoculating wires
A. 8
Dry heat (cont’d)
D. Hot air sterilizer (hot air oven/ dry oven)
It is essential that hot air should circulate between
the objects being sterilized.
These must be loosely packed and adequate air
space to ensure optimum heat transfer.
It is done by applying 140-160 0C for 45 to 60 min
or 180 0C for 30 minutes .
Use: Usually used to sterilize glass wares and
metallic objects.
Physical means…
1.2. Moist heat
It is preferred to dry heat due to more rapid killing
action.
Moist heat can be used by the following methods.
Temperature below 100 0c
• This method does not destroy spores.
- Pasturization: It is the process of application of heat
at temperature of 620c for 30 minutes or 720c for 15
seconds
Uses: Pasteurization of milk ( disinfecting milk) and
Preparation of bacterial vaccines.
Moist heat (cont’d)
Temperatures around 100oC
A. Boiling- (Hot water boilers)
– are still a common methods in many hospitals.
– the maximum temperature is 100oC and will
therefore, not kill all the spores, but for 20 minute
exposure all vegetative forms of bacteria and
viruses can be destroyed provided instruments are
cleaned before putting them in boilers.
Moist heat (cont’d)
Temperatures around 100oC (cont’d)
B. Tyndallization: Intermittent steaming
– Steaming of the material is done at 1000c for 30
minutes for 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.
Moist heat (cont’d)
Temperature above 100oC
Autoclave: Steam under pressure
• It is based on the principle that when microorganism is boiled at
increased pressure (in closed container), hot saturated steam will
be formed which penetrates and gives up its latent heat when it
condenses on objects.
- Hot saturated steam in autoclaving acts as an excellent agent for
sterilization because of:
- High temperature
- Wealth of latent heat (stem under pressure)
- It destroys bacterial endospores and vegetative cells
Uses: Sterilize culture media, gowns, medical and surgical
equipment.
Moist heat (cont’d)
Table: Time-Temperature-Pressure Level Relationship in moist
heat sterilization (Autoclaving)
Temperature Time Pressure level Routinely used
1210c 15 minutes 15 lb/inch2
1260c 10 minutes 20 lb/inch2
1340c 3 minutes 30 lb/inch2
N.B: Most autoclave work by 1210c ,15 minutes and 15 lb/inch2
(temperature, time and pressure respectively).
Abate A. 14
2. Radiation
Gamma rays, x-rays, beta rays, cosmic rays,
ultraviolet light, and even visible light are all forms
of radiation.
When these rays strike an organism, energy may
be absorbed by the cells, often causing cell
damage or death.
Radiation (cont’d)
Uses of radiation:
– Pharmaceutical products such as hormones,
antibodies and enzymes.
– Heat sensitive articles such as surgical suture,
disposable plastic syringes and catheter
– Used to sterilize large room like operation theatre
3. Filtration
• Liquids and gases can be sterilized by passing them
through filters.
• The filter acts as a strainer, a microbial sieve.
• Standard bacteriological membrane filters are
composed of nitrocellulose and have pore diameters
of 0.45 µm, small enough to prevent passage of most
bacteria.
Figure: Micropore filter apparatus
Filtration (cont’d)
Uses of filtration:
Preparing heat sensitive ingredients of culture
media like blood, serum,
Preparing pharmaceuticals and biological solutions
such as vaccines, enzymes, antibiotics, and
injection fluid and IV fluids
Microbial evaluation of water purity
II. Chemical Methods
Classification of disinfectants:
1. Based on consistency
– Liquid (E.g., Alcohols, Phenols)
– Gaseous (Ethylene oxide)
2. Based on mechanism of action
– Action on membrane (Eg., Alcohol, detergent)
– Denaturation of cellular proteins (E.g., Alcohol, Phenol)
– Oxidation of essential sulphydryl groups of enzymes (E.g.,
H2O2, Halogens)
– Alkylation of amino-, carboxyl- and hydroxyl group (E.g.,
Ethylene Oxide, Formaldehyde)
– Damage to nucleic acids (Ethylene Oxide, Formaldehyde)
Abate A. 19
