Sterilisation and Disinfection

Lab 5

Sterilisation and Disinfection

Sterilisation

Killing or removal of all living organisms in or on a substance or object, usually be denaturing proteins, nucleic acids and membrane permeability Killing of most microorganisms on a substance or object (kill may not be complete removes a high % of organisms to numbers too low to cause disease) Procedure carried out in the absence of living things

Disinfection

Sterile techniques

Aseptic techniques

Procedures performed in a way to prevent contamination with infectious microorganisms

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Methods of sterilisation

Heat

Dry hot air oven 160 deg. Moist Autoclave 121 deg. /20 min

Ionising radiation disposable plastics, surgical materials, unsuitable for glass and metal Filtration Chemicals

Ethylene oxide Formaldehyde Hydrogen peroxide Chlorine dioxide Peracetic acid

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Heat - Denatures enzymes

Thermal death point (TDP): lowest temp at which all microorganisms are killed in 10 minutes
Thermal death time (TDT): minimal length of time in which all bacteria will be killed Decimal reduction time (DRT): time, in minutes, in which 90% of a population of bacteria at a given time will be killed Moist heat Coagulation of proteins and breakage of H bonds Boiling kills all vegetative cells, most viruses and fungi within 10 minutes
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Steam under pressure

121C Kills all microorganisms and their endospores in about 20 minutes Monitor efficacy of Autoclave Bacillus stearothermophilus spores

Pasteurization

63C for 30 minutes 72C for 15 seconds (high temperature short-time (HTST) pasteurization ultra-high-temperature (UHT) treatment

Factors affecting the action of disinfection

Period of exposure Concentration Temperature pH Organic matter Number of bacteria present

Disinfection

Chemicals, as well as those used for sterilisation:

Alcohols Dyes -propiolactone Cholorhexidine Hydrogen peroxide Hypochlorites Mercury compounds Phenolics Tetrachlorosalicilanide (TCS)

Variety of mode of action on bacterial cells

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