STERILIZATIO

N AND
ASEPSIS
 INTRODUCTION
Microorganisms are ubiquitous. Since they cause
contamination, infection and decay, it becomes
necessary to remove or destroy them from materials
or from areas. This is the objective of sterilization.

The methods of sterilization employed depend on

the purpose for which sterilization is carried out, the
material which has to be sterilized and the nature of
the microorganisms that are to be removed or
destroyed.
 DEFINITION
Sterilization is defined as the process by
which an article, surface or medium is freed
of all microorganisms either in the vegetative
or spore state.

Sterilization is the total destruction of all

microbes including the more resilient forms
such as bacterial spores, mycobacteria
nonlipid viruses, and fungi.
Disinfection means destruction of all pathogenic organisms,
or organisms capable of giving rise to infection.
Use of physical procedure or chemical agents to destroy
most microbial forms, bacterial spores and other relative
resistant organisms (eg: mycobacteria, nonlipid viruses,
fungi); may remain viable.

Antisepsis
The term antiseptic is used to indicate the prevention of
infection, usually by inhibiting the growth of bacteria.
Chemical disinfectants can be applied safely to skin/
mucous membrane surfaces and are used to prevent
infection by inhibiting the growth of bacteria are called
antiseptics.
 CLASSIFICATION OF INSTRUMENT
STERILIZATION
The categorization of instrument depends on the contact with
different tissue types to determine whether sterilization /
disinfection is required. The categories are as follows.

1. Critical items : Instrument that touches sterile areas of the body

enter the vascular system and those that penetrates the oral
mucosa. Eg : scalpel, curettes, burs
2. Semi – critical items : Instrument that touches mucous membrane
and oral fluids but do not penetrate tissue, Eg.: saliva ejectors,
radiograph, bite blocks, mouth micros etc.
3. Non critical items : Those items that does not come in contact with
oral mucosa but are touched by saliva / blood contaminated hands
while treating patient. Eg.: Chair light, switches, drawer pills etc.
These should be properly disinfected.
INSTRUMENT PROCESSING

Holding solution

Rinse and dry

Instrument cleaning
(ultrasonic)

Rinse and dry

Clinical use

Package

Seal

Sterilize

Store packages
METHODS OF CLEANSING INSTRUMENTS
1. Ultrasonic processing 2. Manual cleaning
Advantage of Ultrasonic processing
Increased efficiency in obtaining a high degree of cleanliness.
Reduced danger to clinician from direct contact with potential
pathogenic microorganisms.
Improved effectiveness for disinfection
Elimination of possible dissemination of microorganisms through
release of aerosols and droplets, which can occur during
scrubbing process.
Penetration into areas of the instruments where the bristles of a brush
may be unable to contact.
Removal of tarnish.

Manual cleansing
Ultrasonic processing is the method of choice, but when manual
cleaning is the only alternative, precaution must be taken to
prevent contamination
 CLASSIFICATION OF VARIOUS
STERILIZATION AGENTS
A) Physical agents B) Chemicals
1) Sunlight 1. Alcohols : Ethyl, isopropyl,
2) Drying Trichlorobutanal
3) Dry heat : Flaming
2. Aldehydes : Formaldehyde
Incineration
Hot air Glutaraldehyde
4) Moist heat : Pasteurization
Boiling 3. Dyes
Steam under normal pressure 4. Halogens
Steam under pressure 5. Phenols
5) Filtration : Candles 6. Surface –active agents
Asbestos pads 7. Metallic salts
Membranes
8. Gases : Ethyleneoxide,
6) Radiation : ultra –violet radiation
Ionizing radiation formaldehyde,
7) Ultrasonic and sonic vibrations beta propiolactone
1) SUNLIGHT

•Sunlight possesses appreciable bactericidal activity

•It is the sterilization which occurs under normal condition.
•The action is primarily due to its content of ultraviolet rays, most
of which, however, are screened out by glass and presence of
ozone in the outer regions of the atmosphere.
•Direct sunlight, as in tropical countryside where it is not filtered off
by impurities in the atmosphere, has an active germicidal effect
due to the combined effect of ultraviolet and heat rays.
•Simple and Grieg showed that, in India, typhoid bacilli exposed to
the sun on pieces of white drill cloth were killed in two hours,
whereas controls kept in the dark were still alive after six days.
Bacteria suspended in water are readily destroyed by exposure to
sunlight.
2) Drying

Moisture is essential for the growth of bacteria. Four-fifth by

weight of the bacterial cell consists of water. Drying in air has
therefore, a deleterious effect of many bacteria. This method
is unreliable and is only of theoretical interest spores are
unaffected by drying.
3) Heat
Materials damageable by heat can be sterilized at lower temperatures, for
longer periods or by repeated cycles.
Moist heat is more effective than dry heat
Moist heat kills microorganisms probably by coagulation and
denaturing their enzymes and structural proteins, a process in
which water participates.
Sterilization ie killing of the most resistance spores, generally
requires moist heat at 121C for 15-30 minute.
Moist heat is effective at lower temperature. A properly
designed autoclave will effectively sterilize media and other
water – containing materials in sealed or unsealed bottles,
laboratory discards and porous packaged materials.
Killing effect of dry heat is due to protein denaturation,
oxidative damage and the toxic effect of elevated levels of
electrolytes.
Dry heat requires a temperature of about 160C for 60 minutes.
Dry heat is suitable for glasswares, instruments and paper-
wrapped articles not spoiled by very high temperatures, and for
water-impermeable oils, waxes and powders. Dry heat cannot
be used for water containing culture media.
a) Flaming / Red Heat
DRY HEAT
b) Incineration
c) Hot air oven
Advantages
More convenient
Instruments are less likely to corrode
Do not require drying after sterilization
Disadvantages
Much harder to kill than when they are wet.
Has very little power of penetration and unless very high temperatures
are used, the method is slow.
Instruments need a considerable time to cool after sterilization and the
temper of metal instruments may be lost.
Used to sterilize glass ware, forceps, scissors, scalpels, all glass
syringes, swabs, measuring cylinders, some pharmaceutical
products such as liquid paraffin, dusting powder, greases.
Sterilization is achieved by dry heat at 160C for 45 minutes, 170C for 15
minutes,180C for 7½ minutes, 190C for 1½ minutes. These exposure
times are considered to be sufficient to kill the spores.
Hot air sterilizer should be fitted with a fan to provide forced air circulation
throughout the oven chamber, a temperature indicator, a control
thermostat, and timer, open mesh shelving and adequate wall insulation.

The oven is usually treated by electricity, with heating elements in the wall
of the chamber. Door interlocks should be fitted to ensure the heating cycle
will not start until the door is shut and that the door will not open while the
cycle is in progress.
The materials should be arranged in a manner which allows free circulation
of air in-between the objects.
The oven must be allowed to cool slowly for about 2hrs before the door is
opened since the glass ware may be cracked by sudden or uneven
cooling.
Moist heat can be employed
At temperature below 100C
At a temperature of 100C (either in boiling water / in free steam) or
At a temperature above 100C (in saturated steam under increased
pressure).

a) Moist heat at temperature below 100C

The best known Eg is in posturisation of milk.
The temperature employed is either 63C for 30min (the holder method) or
72C for 20 sec (the flash method) followed by cooling quickly to 130C or
lower. By these processes all nonsporing pathogens such as mycobacteria,
brucellae and salmonella that may be found in milk are destroyed.
Vaccines of nonsporing bacteria are heat inactivated in special vaccine
baths at 60C for one hour. Serum or body fluids containing coagulable
proteins can be sterilized by heating for one hour at 56C in a water bath on
several successive days.
Human immunodeficient virus (HIV) in serum may require 25 min at 56C for
inactivation.