Sterilization is the process of destruction or removal

of all micro-organisms including the spores.

Disinfection is the process of destruction of

vegetative forms of pathogenic organisms which are
capable of producing infection but not necessarily
resistant spores.

Disinfectant: These are chemical substances which

destroy harmful microbes(usually not spores) on the
surface of an article
Example of disinfectants: Phenol, chlorine, Iodine,
formaldehyde

Antiseptic: These are substances which either kill

micro-organisms or inhibit their growth on the
surface of skin or mucous membrane.
Use:-1. On skin for surgical incision
2. For surgical wounds
Example of antiseptic-Alcohol-70%, Iodophores-1-
2%(povidone iodine), Chlorhexidine
 Methods of sterilization
 Physical methods:
A. Heat:
i) Dry heat-
Red heat
Flaming
Hot air woven
Incineration
ii) Moist heat-
Below 1000C:
Pasteurization
Water bath
 Vaccine bath
Inspissator
At 1000C:
 Boiling
 Steaming-Simple steaming, Tyndalization

Below 1000C:
 Autoclave
B. Radiation:
i) Ionizing Radiation:
 X-Ray
 Y-Ray(gamma Ray)
 β-Ray
ii)Non Ionizing Radiation: Ultraviolet ray
C. Filtration:
i) Earthen ware filter-Berkefeld, Chamberland
ii) Sintered glass filter
iii) Asbestos –Seitz filter
Iv) Membrane filter-Nitrocellulose membrane
v) HEPA(High efficiency particulate air) filter
 Chemical methods
 Alcohols
 Phenols
 Halogens
 Formaldehyde
 Glutaraldehyde
 Ethylene oxide
 Detergents
Dry heat:
Mechanisms-
1. Destructive oxidation or denaturation of essential
cell constituents
2. Toxic effects of electrolytes
Red heat:
Uses- inoculating wires, point of
forceps, needles

Flaming:
Uses- Scalpel blade, needle, glass
slides, mouth of culture tubes and
bottles
Hot air woven:
 It is an electrically heated thermostat controlled
chamber
 Holding times: Hot air woven
 1600C for 60 mins
 1700C for 40 mins
 1800C for 20 mins
Hot air oven : Uses
Sterilization of –
1. Glass wares like syringes, test tubes,
petridishes, flasks, pipettes
2. Sharp cutting surgical instruments like forceps,
scalpels & scissors
3. Oily fluids which are impermeable to water
such as oils and fats
4. Chemicals such as powders which would
clump or form cake in presence of moisture
 Moist heat
Mechanisms
 Denaturation and coagulation of protein in which
water participates
 Membrane damage and leakage of
micromolecules
 DNA damage
 Pasteurization
 Is not sterilization
 It does not destroy spores
 Milk is heated at 630C for 30 min.
 Kills milk-borne pathogens such as:
M. bovis, Brucella, Salmonella, Streptococcus,
staphylococcus, Listeria, Coxiella burnetti.
 Boiling
 Uses: Ryles tube, catheter and other medical
and surgical instruments.
 Method:
1000C for 15-20 min.
 Efficiency: inadequate for sterilization purpose,
many spores withstand.
 Steaming
 Simple Steaming: Exposure at 1000C for 90 min.
Uses: DCA, TCBS, Selenite F broth.

 Tyndallization(Intermittent steaming):
Principle- First exposure at 1000C for 30 min. Vegetative
bacteria are destroyed, but spores remain Incubation
overnight at 370C Germination of spores Second
exposure at 1000C for 30 min. Vegetative bacteria are
destroyed But, few spores may remain Incubation

Germination 3rd exposure All are destroyed.

Uses: To sterilize culture media containing serum, egg,
sugar.
 Autoclave
Sterilization time:
At 1210C under 15 lbs/sq inch for 15-20minutes
 Uses of autoclave
 Linen substances such as operation theatre
musk, gown, cap, draping sheet, bed cover,
aprons
 Dressings
 Blunt surgical instruments: Forceps, Retractors
 Suture materials e.g. silk(except catgut which is
sterilized by ionizing radiation)
 Rubber goods e.g. catheter, gloves, syringes
 Bottle with rubber cap
 Agar based culture media
 Advantages of Autoclave
 As it is moist heat, it has more penetrating power.
 Pressure is applied, so effective under low
temperature
 Liberation of latent heat occurs during
condensation of steam. So, internal temperature
increases
 Less time consuming
 It is one of the best method of sterilization as
bacteria, virus, parasite, fungus including their
spores are killed
 There is no churring of cotton materials
 Disadvantages of Autoclave

 Sharp surgical instruments cannot be sterilized.

Because sharpness is destroyed due to erosive
effect of steam.

 Powder , grease and oily substances cannot be

sterilized as they are not penetrated by autoclave.
 Filtration
Uses: Drugs, vaccines, serum, sugar, water, air
 UV radiation: application

 Disinfection of hospital( particularly operation

theatre)
 Pharmaceutical industries (disinfection of blood
components and derivatives)
 Ionizing radiation
Use: for sterilizing catgut, nylon suture,
disposable needles, syringes and gloves,
transfusion sets, adhesive dressing, tissue
grafts.
 Methods of sterilization that can
destroy bacterial spores
Physical methods: Chemical methods:
 Autoclave  Halogen(chlorine, Iodine)
 Hot air oven  Formaldehyde
 Tyndallization  Glutaraldehyde
 Ionizing radiation  Ethylene oxide
 Gas plasma sterilizer
 Peracetic acid