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Sterilization Methods

Ranjith Kumar Inbasekaran M.Sc Renal Sciences and Dialysis Technology SRU Joseph Lister

Sterilization
Free of all microorganisms, either in the vegetative form or Spore state

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Microbiology/ Sterilization Methods

Descending Order of Resistance


Prions Bacterial Spores Mycobacteria Non Lipid or Small Viruses Fungi Vegetative Bacteria Lipid or Medium Size Viruses
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Ideal Sterilization Properties


1. Effective 2. Safe 3. Fast 4. Penetrative 5. Removable 6. Monitorable 7. Cost effective 8. Adaptable 9. Maintainability 10 Compatibility

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Microbiology/ Sterilization Methods

Sterilization Process (Physical)


Sunlight Drying Heat Radiation Filtration Pressure release Plasma Ultrasonic and Sonic Vibrations
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Sun Light
Natural Source Presence of UV rays and Heat Rays

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Drying
Absence of moisture Removal of water from bacterial cell Not effective

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Microbiology/ Sterilization Methods

Heat
Dry Heat Moist Heat - Flaming -Pasteurization - Incineration -Inspissator - Hot air Oven - Boiling - Steam with Pressure -Steam without Pressure

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Microbiology/ Sterilization Methods

Flaming
Inoculation loop or Wire, the tip of Forceps and spatulas are held in a bunsen flame till they are red hot.

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Incineration
This is an excellent method of destroying materials such as contaminated cloth, animal carcasses and pathological materials.
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Hot Air Oven


Kills by oxidation effects Dry heat sterilize articles Operating Temp 50oC to 250/300oC. Holding time 160oC for 1 hr

Thermostat controls temp.


Double walled insulation. Forceps, Scissors, Scalpels, Liquid paraffin, fats and grease
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Pasteurization
Process of killing of pathogens in the milk but does not sterilize it . Milk is heated at 63oC for 30 mins. (Holder Method) At 72oC for 15-20 Sec. Rapid cooling to 13oC (Flash Method) Mycobacteria, salmonella etc are destroyed
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Inspissator
Sterilizes by heating at 80-85oC for half an hour for 3 successive days Used to sterilize media such as Lowenstein-Jensen & Loefllers serum
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Boiling
Kills vegetative forms of bacterial pathogens. Hepatitis virus: Can survive up to 30 minutes of boiling. Endospores: Can survive up to 20 hours or more of boiling
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Steam Sterilizer (Atmospheric Pressure)


Steam is generated using a steamer (Koch/ Arnold) Perforated tray above ensures materials are surrounded by steam. For routine sterilization exposure of 90 mins is used
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Steam Sterilizer Under Pressure


Charles Chamberland Auto clave.

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Autoclave Machines- Types


Downward Displacement Vacuum Assisted Pulse Steam Dilution

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Advantages
Fast Penetration of loads Complete Sterilization No Residual Toxicity No Damage Easy quality control Economical Reliable
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Disadvantage
Not suitable for heat sensitive materials Not suitable for non permeable materials Potential for burns

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Autoclave
Sterilization Time (Holding time +Safety Time) Pressure Temperature

2+1 min 8+2 min 12+3 min

30 Psi 20 Psi 15 Psi

134 C 126 C 121 C

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Autoclave

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Radiation
Ionizing - X Ray - Cathode - Gamma Non Ionizing - UV
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Radiation
Source: Cobalt 60 / Caesium 137/ Electron Beam Reliable Good penetration No residual activity at 2.5 mega Rhontgens Glass becomes dark, cotton losses tensile property
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Radiation

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Filtration
Candle Filters Asbestos filters Sintered glass filters Membrane filters

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Ultrasound

More effective for gram-negative bacteria Lack of thoroughness - survivors remain

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Sterilization Validation
Effective sterilization process- a spore kill to achieve SAL (106)

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Time
171 C, 60 minutes, Dry heat 160 C, 120 minutes, Dry heat 149 C, 150 minutes, Dry heat 141 C, 180 minutes, Dry heat 121 C, 12 Hours, Dry heat 121 C, 15 minutes, Moist Heat With Pressure
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Thank You

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References
Microbiology by Ananthanarayanan and Panicker F.T Sanders Sterilization and Disinfection http://generalbacteriology.weebly.com/steriliz ation-and-disinfection.html http://lpc1.clpccd.cc.ca.us/lpc/zingg/Micro/lect s.htm
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