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Sterilization & Disinfection Physical Agents. Hugh B. Fackrell PhysSter.ppt. Sterilization & Disinfection. Definitions Bacterial Cell Death Physical methods Heat Radiation Filtration. Definitions. Sterilization Complete Destruction of ALL microbes Disinfection
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Sterilization & DisinfectionPhysical Agents Hugh B. Fackrell PhysSter.ppt
Sterilization & Disinfection • Definitions • Bacterial Cell Death • Physical methods • Heat • Radiation • Filtration
Definitions • Sterilization • Complete Destruction of ALL microbes • Disinfection • Killing of pathogens on inanimate objects • physical or chemical • Antiseptic • A chemical agent for disinfection of living tissue • External skin, mouth vagina
Definitions • Bacteriocide • kill microbes • also germicide, fungicide, virucide • Bacteriostatic • Prevents or stops microbial growth • also fungistatic, virustatic • Aseptic(Asepsis) • Prevent contamination of person or object by microbes
Definitions • Sanitize • Removal of pathogens from inanimate objects • Mechanical or chemical cleaning • need not sterilize of disinfect • Contamination • Presence of living microbes on object
Definitions • Infection • Presence of living multiplying microbes in host tissues • often pathogenic
Survivors (log) 1 log decrease = 90% kill 5 1 log Death rate (D) varies with bacteriocide 4 3 2 D 1 Time(min) to kill 90% 0 Time (min) Bacterial Cell Death
Physical Methods • Heat • Radiation • Filtration
Heat Sterilization • Factors • length of exposure • temperature • “Moist Heat” • “Dry Heat” Method of choice
“Moist” heat • Effective quickly against most pathogens • low thermal death point • 60-70% in 10 min • Pasteurization 60-70 oC Sanitizes • Hot water 60-100 oC Disinfects • Steam 15 psi 121oC Sterilizes Endospores resist boiling for hours!!
Heat: Mechanism of Action • Inactivation of enzymes • Structural changes • coagulation of proteins • Oxidation • membrane disruption
Pasteurization • Liquids • Minimal heat • 145 oF (62.9 oC) - 30 min • 162 oF (71.6 oC) - 15-30 sec • Flavour of drink preserved • Kills non spore forming pathogens • TB. Brucella, Salmonella, Streptococcus • Kills spoilage bacteria • Lactobacillus, Streptococcus
“Flash” Pasteurization • 72 oC - 15 sec • 88 oC - 1 sec • 90 oC 0.5 sec • 96 oC 0.05 sec Raw milk Heating coil Cooling coil
Boiling Water • 100 oC (212 oF) 10- 30 min • Disinfectant • spores & some viruses are resistant • Safe for clean instruments that DO NOT penetrate the tissue • Soiled items -add detergent
Steam Under Pressure • Autoclave • 15 psi 121 oC 250 oF • Time 15 min - dependent on volume • Steam displaces air & penetrates • Packing of autoclave critical! • Airlocks in packing • reduce effective temp & pressure
Dry Heat • Longer & hotter • Oxidation • Carbonization • cotton plugs
Types of Dry Heat • Oven • 160-165 oC 2 hours • 170-175oC 1 hr • Glass, oils, waxes, glycerin • Incineration • destruction of pathological material • Flaming • used as aseptic techniques • scissors, scalpels, loops, needles
Radiation Sterilization • Ionizing Radiation • Ultraviolet Radiation * Point mutations involve changes in only one or a few nucleotides
Ionizing Radiation Increased energy Radiant Energy spectrum UV light
Sterilization by Ionizing Radiation • Gamma Rays • Food • Plastics • Electron Beams • pharmaceuticals Human Hazard!
Ionizing Radiation • Creates free radicals • Causes breaks in two DNA strands • Results in deletions • Results in translocations
Free radicals can breaks both phosphodiester bonds of a DNA helix Free Radicals • High Energy Radiation • energy absorbed by atoms • energy added to electrons • electrons ejected from atoms • leaves ionized atoms & unpaired electrons
Sterilization by Ultraviolet light • Circulating air • Surfaces • benchtops • Small samples of liquid Surfaces only poor penetration Photoreactivation white light reverses effect of ultraviolet
Ultraviolet Radiation • Pyrimidine dimers • errors during repair • Peroxides, Free radicals • See ionizing radiation • Direct molecular damage • intense exposure
TT GTA GTC CATAACAG TT A GT GTC CATAACAG TT A G GT GTC CATAACAG GTA T GTC CATAACAG GTA TTGTC CATAACAG Repair of thymine dimer
Filtration Sterilization • HEPA filters • High Efficiency Particulate Air filter • Remove airborne microbes • Filters particles >0.3 mm • Operating rooms • “Clean rooms” instrument assembly • Burn patients • Immunocompromized
Filter sterilization • Membrane filters • cellulose nitrate or synthetic polymers • Pore size • bacteria 0.22mm 0.45 mm • viruses 0.01 mm • Sterilize heat or energy sensitive materials • bacterial media • drug solutions