STERILIZATION,

DISINFECTION AND
ANTISEPSIS V.Snegarova-Toneva M.D
Department of Microbiology and Virology,Medical Faculty -Varna
Aim of sterilization:Asepsis
◦ Asepsis is the practice to reduce or eliminate contaminants
(bacteria,viruses,fungi,parazites) from entering the operating field in surgery
or medicine to prevent infection.
◦ Ideally ,a field is sterile-free of contaminants
◦ This aim is difficult to achieve
◦ However the aim is to eliminate infection of the patient ,not absolute sterility
 Sterilization
Sterilization is the total destruction of all microbes,including the more resilient
forms such as bacterial spores,mycobacteria,nonenveloped(nonlipid)viruses and
fungi.
A surface or an object is either sterile or not sterile,there is no gradations of
sterility!
There are 3 main ways of sterilization:physical sterilants,gas vapor
sterilants,chemical sterilants
 Physical sterilants
Physical sterilization:dry heat and moist
◦ Dry heat sterilization is the most common way in hospitals indicated for most materials.
The simple flaming of Bunsen lamp of the loop fot 5-7 sec under 1560 C,until red
1 Dry heat sterilizator(Pasteur sterilizator,hot air oven) to have good effect we need more
time and higher temperature than with moist ,double wall insulation with thermostat
160-180 C for 1-2 hours
Cheap and easy way
NB!!glass instruments and all laboratory utensils,scissors,scalpels,swabs,paraffin ,fats always
put well cleaned and dry
Risk of breaking and bad sterilization if opened before temperature 40-50 degrees
 Physical sterilization

◦ 2 Sterilization with moist

The oldest way (1881)with boiling water 100 degrees,but some spores and viruses don’t die it’s
used for clothes,shoes,linen,gown and personal belongings if no other choice
Pasteurization-named of the famous chemist and microbiologist Louis Pasteur in 1862,this
method is used nowadays for liquids reach of proteins and vitamins like milk,juices,wine,which
lose their quality under high temperatures,70-80C for 15 min,kills food spoiling and labile
pathogens but not spores
Tindall method(English physist 1876)-fractional sterilization by intermittent heating for sugar
media,liquids like serums,medications,several days under 50-60 degrees for 20-45 min,vegetative
forms die and spores vegetate between heatings
 Physical sterilization

◦ Robert Koch non pressure flowing steam sterilizer(Koch’s sterilizer)

introduced 1881
 Physical sterilization
◦ Koh’s sterilizer-uses steam atmospheric pressure
Steam is generated by steamer
Consists of a tin cabinet
Has a conical lid to assure the drainage of condensed steam
Perforated tray above ensures that materials are surrounded by steam
90 min for routine exposure! Vegetative forms die but some spores
survive
 Physical sterilization
Autoclave
This is a high pressure device used to allow the application of
moist heat above the normal athmosphere boiling point of water
Exposure to 121C for 15+ min at 1 atm is typically sufficient
Large items,large volumes and items poorly penetrated by
steam take much longer to sterilize
The best method for sterilization-even for spores
 Physical sterilization
Radiation sterilization(cold sterilization)-radiation ionizes
water and turns it into an oxidizing agent,it has different degrees
of penetration
Non ionizing radiation-infrared,UV light,ultrasound and
radiofrequent radiation
Ionizing radiation(DNA targeting)-particulate
radiation(accelerated electrons),high energy gamma rays and Re
rays
 Filtration sterilization
◦ This method does not destroy the microorganisms but removes them
◦ This is a mechanical way of sterilization
◦ Capable of preventing the passage of viable and non viable particles
◦ Used for purifying and sterilization of liquids and gases
◦ The major mechanisms of filtration are sieving,adsorption and trapping within
the matrix of the filter material e.g. HEPA filters
◦ Sterilization of ophthalmic solutions,biological products,heat sensitive i.v.
medications
◦ Part of machines like centrofuges,autoclaves,freeze driers,fermenters etc
 Sterilization vs Disinfection
◦ Disinfection and sterilization are terms casually interchanged and this causes
confusion
◦ Disinfection could be :high,intermediate and low level
◦ High level disinfectants approach sterilization in effectiveness,used for items
involved with invasive procedures that cannot withstand
sterilization(endoscopes,surgical instruments with plastic parts)
◦ Remove all organic residue!!
E.g. glutaraldehyde,hydrogen peroxide,peracetic acid,chlorine
 Disinfection
◦ Factors influencing their activity:
Concentration and kind of an agent used
The length of exposure to the agent
The temperature
The kinds of microorganisms present
The number of microorganisms present
The nature of material bearing the microorganism
 Sterilization vs Disinfection
◦ Intermediate-level disinfectants(alcohols,iodophor compounds,phenolic
compounds)
◦ Used for instruments and surfaces where spores and highly resilient organisms
are exception
◦ These instruments are semicritical :flexible endoscopes,laryngoscopes,vaginal
specula,anesthesia breathing circuits
◦ Low level disinfectants-(quaternary ammonium compounds)
◦ They are used for noncritical instruments like blood pressure
cuffs,electrocardiogram electrodes,stethoscopes
 Antisepsis
◦ Antiseptic agents are used to reduce the number of microbes on skin surfaces.These
compounds are selected for their safety and efficacy.
◦ Alcohols:
◦ excellent activity(but not spores!)
◦ Nontoxic,but tend to dry the skin,because they destroy lipids
◦ They don’t have residual activity and are inactivated from organic matter
◦ Iodofors
◦ More toxic,allergy
◦ Inactivated from organic material,limited residual activity
 Antisepsis
◦ Chlorhexidine
◦ Broad antimicrobial activity
◦ It’s activity persists longer,organic material decreases it
◦ Parachlorometaxylenol-only Gram positive bacteria
◦ Non toxic,high residual activity
◦ Used in handwashing products
◦ Triclosan-active against bacteria but no other
◦ Used in deodorants and toothpaste
Desinfection