STERILIZATION

AND
DISINFECTION
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Microorganisms are the agents of contamination
and infection.

Hence it becomes necessary to remove them from

materials and areas.

Early civilization practiced salting, smoking,

pickling and exposure to sunlight.

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Definition of terms
Cleaning: The physical removal of organic
material or soil from objects, is usually done by
using water with or without detergents.

Sterilization: It is a process by which articles are

freed of all microorganisms both in vegetative as
well spore state.

Disinfection: It is a process of destruction of

pathogenic organisms capable of giving rise to4
infection.
Important: First Objects to Be Sterilized
Should Be Cleaned by Washing

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APPLICATION OF DISINFECTANTS

In Bacteriology

- For disposal of culture (Lysol).

- For preservation of agar or phenol agar,
vaccine, etc.
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In Surgical Procedures
- Washing the hand.
- To prepare and clean the area of operation.
- To collect the blood under aseptic precautions.
- For safe disposable of excreta and surgical
dressing.
- Cleaning of infected wounds. 7

- For disinfection of used instrument.

In Hospitals

- To disinfect the operation theaters.

- To disinfect costly equipments like
endoscopes and cystoscopes, etc.
- To control the spread of cross infection.
- To disinfect linen and surgical dressing.
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 In Public Health Services

- For providing safe drinking water (e.g.

chlorinated water).
- For disinfection of sewage before its disposal
into the fields.

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Antiseptic: It means prevention of infection by
inhibiting growth of bacteria.

Bacteriocidal agents: These are those which are

able to kill bacteria.

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 Physical Methods

Sunlight: It possesses appreciable bacteriocidal

activity. The action is due to ultraviolet rays.
This is one of the natural methods of sterilization
in case of water in tanks, river and lakes.

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Drying: Drying in air has deleterious effect on
many bacteria. Spores are unaffected by drying.
Hence, it is a very unreliable method.

Heat: The factors influencing sterilization by

heat are:

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 Dry Heat
Killing by dry heat is due to:

1- Protein denaturation.
2- Oxidative damage.
3- Toxic effect of elevated levels of electrolytes.

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A- Red Heat: It is used to sterilize metallic
objects by holding them in flame till they are red
hot, e.g. inoculating wires, needles, forceps, etc.

B- Flaming: The article is passed over flame

without allowing it to be red hot, e.g. mouth of
culture tubes, cotton-wool plugs and glass slides.
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C- Incineration: This is an excellent method for
rapidly destroying material, e.g. soiled dressing,
animals carcasses, bedding and pathological
material, etc.
D- Hot air oven: Sterilization by hot air oven
requires temperature of 160°C for one hour. We can
sterilize all glass syringes, petridishes, test tubes,
flask, pipettes, cotton swabs, scalpel, scissors,
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liquid paraffin, dusting powder, etc.

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 Sterilization Control of Hot Air Oven

1- The spores of non-toxigenous strain of

Clostridium tetani are used to test dry heat
efficiency.
2- Browne’s tube (green spot) is available for
sterilization by dry heat. A green color is
produced after 60 minutes at 160°C.
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 Moist heat

The lethal effect of moist heat is by denaturation

and coagulation of protein.

Temperature below 100°C

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1- Pasteurization of milk: Temperature employed
is either 63°C for 30 minutes (holder method) or
72°C for 15 to 20 seconds (flash method). Heating
is always followed by sudden and instant cooling
of milk. Organisms like Myco-bacterium,
Salmonellae and Brucellae are killed, Coxiella
burnetii is relatively heat resistant and hence, may
survive the holder method. 21
2- Vaccine bath: It is used for killing non-sporing
bacteria which may be present in vaccine. In
vaccine bath the vaccine is treated with moist heat
for one hour at 60°C.

3- Inspissation: The slow solidification of serum

or egg is carried out at 80°C in an inspissator, e.g.
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serum slopes, Lowenstein-Jensen’s medium, etc.
 Temperature at 100°C
1- Tyndallization: This is a process by which
medium is placed at 100°C in flowing steam for 30
minutes each on 3 successive days. The
mechanism underlying this method is that
vegetative cells get destroyed at 100°C and
remaining spore which germinate during storage
interval are killed on subsequent heating. 23
2- Boiling: Most of vegetative form of bacteria,
fungi and viruses are killed at 50 to 70°C in short
time. For needles and instruments boiling in water
for 10 to 30 minutes is sufficient to sterilize them.
Addition of little acid, alkali, or washing soda,
markedly increases the sterilizes power of boiling
water. Spores and hepatitis viruses are not readily
destroyed by such procedure. 24
3- Steam at atmospheric pressure (100°C):

here free steam is used to sterilize culture

media which may decompose is subjected to
higher temperature. A Koch or Arnold steamer
is used. This is a cheap method of sterilization.

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Temperature above 100°C

Steam under pressure: For bacteriological and

surgical work boiling is not sufficient because
spores survive boiling. Hence, high pressure
sterilizer or autoclave is used.

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 AUTOCLAVE
In this apparatus, material for sterilization is
exposed to 121°C for 15 to 20 minutes at 15 lb
pressure per square inch. Saturated steam heats the
article to be sterilized rapidly by release of latent
heat.

Autoclave is used for culture media, rubber goods, syringes,

gowns and dressing, etc. 27
 Sterilization control

1- Bacillus stearothermophilus.
2- Browne’s tube.
3- Autoclave tapes.
4- Automatic controls or pen records.
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 Types of Autoclave

1- Simple iron jacketed.

2- Low pressure/low temperature.
3- High pressure high vacuum type having facility
to expel 98 percent of air rapidly by an electric
pump and hence sterilization is done quickly.

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 Sterilization by Filtration

This is a method of sterilization useful for

antibiotic solutions, sera, carbohydrate
solution, etc. We may also get bacteria free
filtrates of toxin. It is also useful when we
want to separate microorganisms which are
scanty in fluid.
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 Other Uses of Filter

- Separation of soluble products of bacterial

growth, e.g. toxins.
- Sterilization of hydrated fluid.
- Sterilization of serum.
- Sterilization of antibiotic solution.
- Sterilization of blood products.
- Purification water. 31
Membrane filter:

They may be comprised of cellulose esters. They

can be used as under:
1- Water analysis
2- Sterility testing of solution
3- Preparation of parental solution.
Nitrocellulose membrane filter is commonly used.
It is also called multipore. Membrane
filters are available in pore sizes of 0.015 µm,
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0.12 µm and 0.22 µm.
Vacuum Pump for Filtration

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 Radiations

A- Ultraviolet radiations:
It is chief bacteriocidal factor present in sunlight.
Commonly used UV lamp is of low pressure
mercury vapor type whose length is 253.7 mm. It
causes following changes is cell:

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1- Denaturation of protein.
2- Damage to DNA.
3- Inhibition of DNA replication.
4- Formation H₂O₂ and organic peroxide in
culture media.

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Ultraviolet lamps are used in:
A- Killing of organisms.
B- Making bacterial and viral vaccines.
C- Prevention of airborne infection in operation
theater, public places and bacteriological
laboratories.

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B- X-rays and other ionizing radiations:
Ionizing radiations have greater capacity to
induce lethal changes in DNA of cell. They are
useful for the sterilization of disposable material
like disposable syringes, adhesive dressing, etc.

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C- Gamma radiations
X-ray are utilized using two types of mechanics.
1- Linear accelerator for X-ray.
2- Cobalt-60 for gamma rays.

A dose of 2.5 M rad is sufficient to kill both

vegetative and spore form of bacteria. They
are used to sterilize rubber or plastic syringes,
surgical catgut, bone tissue graft, adhesive dressings,38
etc.
Common Radiation Unit

1 Gy = 100 rad or
1 rad = 0.01 Gy
1Mrad = 10 kGy

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Cobalt 60 for Sterilization

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Cobalt-60(Co60) Radiation Source

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What can be Sterilised by gamma irradiation?

• Medical devices.
• Pharmaceuticals.
• Combination drug/device products.
• Tissue-based and biological
products.
• Animal retail products.
• Cosmetics and toiletries.
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 Chemical Methods
Following chemicals are of common use:

1- Acid and alkalies: They are inhibitory to the

growth of bacteria. Mycobacteria are more
resistant to acid than alkalies. Boric acid is weak
antiseptic.

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2- Halogens: Three of the halogens, i.e. chlorine,
iodine and bromine are among the best
bacteriocidal agents.

Halogens kill vegetative bacteria, fungi, viruses

but not tubercle bacillus or bacterial spores.

Iodine is used chiefly for skin.

Chlorine combines with water to form

hydrochloric acid which is bacteriocidal.
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3- Oxidizing agents: They are weak antiseptic.
e.g. H₂O₂, potassium permanganate.

4- Formaldehyde: It is useful in sterilizing

bacterial vaccine and in inactivating bacterial
toxin without affecting their antigenicity. 5 to 10
percent solution in water kills many bacterial. It is
bacteriocidal, sporicidal and lethal to viruses also
uses:

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1- Disinfection woolen blankets, wool hides
to destroy bacterial spores.
2- Footwear of person with fungal infection
(athletic foot).
3- Wards and operation theater.

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5- Phenol: It is used for sterilizing surgical instruments,
and for killing culture accidentally split over in the
laboratory. It is generally used in 3 solution.

6- Soap and detergents: They are bacteriocidal and

bacteriostatic for Gram-positive and some acid-fast
organisms. Detergent acts by concentrating at cell
membrane and thus disrupting its normal function or it
may denaturate protein and enzyme.

7- Alcohol: Ethyl alcohol is most effective in 70 percent

solution than 100 percent alcohol. It is so because 70
percent alcohol has better penetration power than 100 48
percent alcohol. It does not kill spores.
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Uses of Disinfectant:
In practice disinfectant is useful and necessary for:

1- Contaminated disposable material before

incineration.
2- Surfaces like table and trolley top.
3- Cleaning material when contaminated material
has been split.
4- Disinfection of instruments, not amendable to
heat.
5- Disinfection of skin. 51
Cleaning Disinfection and
Sterilization in Dentistry
Practice

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- Spread of microbes between patients and Dental
team is to be prevented.

- Especial attention should be given to

instruments decontamination together with
surface disinfectant.

- Key notes of infection control involves

cleaning, disinfection and sterilization.
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- Instruments must be rinsed after cleaning with
clean water to remove, chemical residue.
Following rinsing instruments must be dried
using disposable paper towels.

- Disinfection using moist heat is the first choice. It

does not leave any toxic residue on devise
processed.

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- Other techniques like disinfection by some
liquid chemicals, ultraviolet radiation,
filtration and gases are not in much use in
dentistry practice.

- Common disinfectants used in dental practice

includes:
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1- Ethyl, isopropyl alcohol:
A- It required at least 3 minutes contact to
achieve disinfection.

B- It is not sporicidal.

C- It is bacteriocidal, fungicidal and veridical as

well.

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2- Quanternary ammonium compounds
(Cetylpridinium Chloride):
A- It is inactivated quickly by organic matters.

B- It is incompatible with soaps.

C- It has poorly activity against environmental

Gram-negative bacteria.

D- Not sporicidal.
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3- Halogens (Chlorine containing agents):

A- Extremely active.
B- It is bactericidal and virucidal.
C- Inactivated by organic matter.
D- Not sporicidal.

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