STERILISATION AND DISINFECTION

OBJECTIVES
Knowledge of what is sterilisation
Knowledge of what are the different methods of sterilisation the principle and procedure of each, their applications and limitations

Sterilization
Sterilization: is a process by means of which an article,surface or medium is rid of all living microorganisms including spores Disinfection:is a process of destruction of vegetative forms of pathogenic organisms which are capable of producing infection but not necessarily resistant spores.

Sterilization
Physical agents Sunlight -uv rays and heat has germicidal activity Drying - spores are unaffected Heat - most reliable, certain and rapid method of sterilisation Filteration radiation

Chemical agents
Phenols and cresols Halogens Metallic salts Aldehydes Alcohol Dyes Vapour-phase disinfectants Surface- active disinfectants

Sterilization
Red Heat - inoculating loops and wires, points of forceps and spatulas Flaming- scalpel blades, needles mouth of culture tubes, glass slides and cover slips
Incineration rapidly destroys contaminated material i.e. soiled dressings Hot air oven

Hot air oven

Principle: eg of dry heat -killing effect is due to protein denaturation, oxidative damage and toxic effect of elevated levels of electrolytes Used for glass ware like glass syringes, test tubes, petri dishes, pipettes, and flasks, metal instruments, such as forceps, scissors and scalpels Oils, jellies and powders and swab sticks packed in test tubes. Holding time at 160o C 1 hour

Hot Air Oven contd

Precautions: should not be overloaded, substances to be sterilised should be absolutely dry Oven should be allowed to cool for 2 hours before opening to prevent cracking of glassware articles like rubber goods, fabrics,inflammable or volatile substances should not be placed inside

DRY HEAT STERILISATION-

Hot air oven

HOT AIR OVEN

Sterilization by moist heat

At temp below 100oC At a temp of 100oC Boiling Free steam At temp above100oC

At temperature below

o 100 c

Heat labile fluids may be disinfected PASTEURIZATION : *holder method 63oc for 30 minutes *flash method- 72oc for 20 seconds followed by rapid cooling at 13 oC By this method nonsporing organisms like mycobacteria, brucella,and salmonellae are destroyed

Coxiella burneti survive Heat labile fluids such as serum may be disinfected by heating at 56oc
Vaccines may be inactivated in a water bath at 60oc for one hour Household utensils and patients clothings may be disinfected by washing at 70oc-80oc

Moist heat sterilisation at

o 100 c

Boiling at 100oc for 10 to 30 min- at this temp vegetative bacteria and some spores are killed Metals glass and rubber items are boiled, dried cooled and used.

Free steam at

o 100 c

Steam at normal atm pressure is at 100oc It has latent heat which on condensing on the article to be sterilised releases its latent heat. One single exposure to steam for 90min ensures complete sterilization but for media containing sugar and gelatin

cont
An exposure of 100oc for 20 minutes on three consecutive days TYNDALLISATION First exposure to steam kills all vegetative bacteria and any spores present will germinate and will be killed on subsequent occasions

Moist heat at temp above

o 100 c

It provides greater lethal action of moist heat It is quicker in heating up the exposed articles It can penetrate easily porous material such as cotton wool stoppers, paper,and clothwrappers, bundles of surgical linen and hollow apparatus.

AUTOCLAVE
Principle: water boils when its vapour pressure equals that of surrounding atmosphere. Hence when pressure inside a closed vessel increases the temperature at which water boils also increases. Saturated steam has greater penetrating power.When steam comes into contact with a cooler surface it condenses to water and gives up its latent heat to that surface.

Autoclave
Sterilisation by steam under pressure (autoclaving) is suitable for culture media and aqueous solutions, dressing materials, linen, gloves etc Satisfactory sterilisation can be achieved at 15 pounds pressure per square inch at 121oc for 20 min

AUTO CLAVE

AUTOCLAVE cont

Sterilization controls
Biological control Bacillus stearothermophilus Bacillus subtilis Chemical control Brownes tube

Sterilization controls
Chemical controls Biological indicators

Filtration
Liquids such as sera and heat labile substances such as sugars and urea used for media preparation. Separation of bacterial toxins from bacteria Isolation of organisms which are scanty in fluids. Viruses are too small so they cannot be filtered

Filtration contd
Earthen ware filters Asbestos filters (Seitz) Sintered filters Membrane filters Syringe filters

BACTERIAL FILTERS

Seitz filter

Sintered glass filter

Membrane filter

Radiation
Non ionizing UV rays- in the range of 250 to 260 nm wavelength Low pressure mercury vapour lamp Spores are resistant to radiations Radiations induce interference in the DNA replication

Cold sterilization
Ionizing X-rays Gamma rays Cosmic rays High penetrative power lethal to bacteria Prepacked disposables such as plastic syringes, transfusion sets, catheters, canulas, culture plates that are unable to withstand heat.