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By S. B. Sangavi (2011 Batch) Department of Microbiology Dr. Sophia Dr. Padmini Mrs.

Kanchana devi


A process by which an article, surface or medium is made free of all micro-organisms either in the vegetative or spore form. Disinfection: The destruction of all pathogens or organisms capable of producing infections but not necessarily spores. Antiseptics: The chemical disinfectants that can be safely applied to the skin or mucous membrane are called as antiseptics.

Agents Of Sterilisation
Agents of sterilisation

Physical agents

Chemical agents





Alcohols Aldehydes Phenols Halogens Oxidising agents Salts

Dry Heat

Moist Heat

Surface active agents Dyes Gases

> Sunlight has an active germicidal effect due to its contents of ultraviolet rays. > It is a natural method of sterilisation in case of water in tanks, rivers & lakes. > It cannot be used in sterilising medical instruments

> Heat is the most reliable and commonly employed method of sterilisation. > Two types of heat are used dry heat and moist heat. Principle: > Dry heat - denaturation of bacterial protien, oxidative damage and by the toxic effect of elevated levels of electrolytes. > Moist heat - denaturation & coagulation of protiens.

Dry Heat Sterilisation


Red heat: Inoculating wires or loops, tips of forceps and needles are held in the flame of a Bunsen burner till they become red hot. Flaming: Glass slides, scalpels and mouths of culture tubes are passed through Bunsen flame without allowing them to become red hot. Incineration: By this method infective material of no more use is reduced to ashes by burning in an incinerator. Soiled dressings, animal carcasses, bedding & pathological materials are dealt with this method.

Hot Air Oven

It is the most widely used method. The oven is electrically heated and is fitted with a fan. It is also fitted with a thermostat.

160 :C and a holding time of about 2hrs.

Hot Air Oven

Uses: It is used for the sterilisation of Glasswares like glass syringes, flasks, test tubes, petridishes and pipettes Surgical instruments like scalpels, scissor, forceps etc. Chemicals such as liquid paraffin, fats, sulphonamide powder etc.

Hot Air Oven

Precautions: It should not be overloaded. The oven must be allowed to cool for atleast 2 hrs.

Hot Air Oven

Sterilisation control: Biological indicator- The spores of non-toxigenic strains of Clostridium tetani are kept inside the oven. The spores will get destroyed if the sterilisation is proper. Thermocouples are used to record the temperature directly by a potentiometer Brownes tube with a green spot is available.

Moist Heat Sterilisation

At a temperature below 100:C

Moist Heat Sterilisation

At a temperature of 100:C

At a temperature above 100:C

At A Temperature Below 100:C

Pasteurisation of milk: By two methods a) Holder method- 63:C for 30 min b) Flash method- 72:C for 20 s followed by cooling quickly to 13:C or lower

Inspissation: Sterilisation of Loefflers serum or egg media by heating at 80 85:C for half an hour daily on three consecutive days using an inspissator. Vaccine bath: Bacterial vaccines at 60:C for one hour.


At A Temperature Of 100C
Boiling: For 10 30 min. Many spores may withstand boiling for a considerable time. Used for sterilising glass syringes & rubber stoppers. Tyndallisation or intermittent steam sterilisation: Steam at 100:C for 20 min is used. Principle- The first exposure kills all the vegetative forms, in the intervals between the heatings the remaining spores germinate to vegetative forms which are killed on subsequent heating. Use- to sterilise egg, serum or sugar containing media. Steam steriliser: at 100:C for 90 min. Kochs or Arnolds steam steriliser

At A Temperature Above 100C ( Under Pressure )

Principle: When steam comes in contact with a cooler surface it condenses to water and liberates its latent heat to the surface. Components of autoclave:
Asbestos washer Pressure gauge Lid Discharge tap Safety valve Screw clamp


Working Of Autoclave

At 121:C and 15 pounds per square inch for 15 minutes

Uses: To sterilise culture media, dressings, gloves, gowns, rubber materials etc. Sterilisation Control: Thermocouple Bacterial Spores: Spores of Bacillus stearothermophilus are used as the test organism. Chemical Indicators: Brownes tube contains red solution which turns green at 121:C for 15 min in an autoclave Autoclave tapes


Cholera vibrio

Used for substances which get damaged by heat process. Eg. Sera, sugars, antibiotic solutions etc. Uses: Separation of toxins and bacteriophages from bacteria. To obtain bacteria free filtrates of clinical samples to isolate viruses. Salmonella typhi Purification of water. Types of filters: Earthenware candles Asbestos disc filters (Seitz filter) Sintered glass filters Membrane fiilters

Types Of Filters
Candle filters:
a) b) Diatomaceous earth filters eg. Berkfeld Unglazed porcelain filters eg. Chamberland

Types Of Filters
Asbestos disc filters (Sietz filters) :

Types Of Filters
Sintered glass filters:

Types Of Filters
Membrane filters: Made up of cellulose esters. Nitrocellulose membrane filters are widely used.

Two types of radiations Ionising & non ionising radiations. Ionising radiation: (cold sterilisation) Gamma rays, X- rays & cosmic rays. Gamma rays disposable items such as plastic syringes, swabs, culture plates, catheters etc.

Non-ionising radiation: Infra-red & ultraviolet radiation. IR rapid mass sterilisation of syringes & catheters. UV rays 240 280 nm have bactericidal activity. UV rays bacteriological laboratory, operation theatres. Spores are highly resistant to UV rays.

Stainless steel instruments & mirrors any accepted method Handpieces autoclaved Burs dry heat Metal impression trays all accepted methods

Towels autoclaved
Choosing a reliable sterilising equipment of proper size & cycle

compatible with needs of practice is essential.

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