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Microbiology: Principles and Explorations
Sixth Edition
Chapter 12:Sterilization and Disinfection
Copyright © 2005 by John Wiley & Sons, Inc.
Jacquelyn G. Black
Principles of Sterilization and Disinfection
• Disinfectant: Typically chemical agents that are applied to inanimate objects
• Antiseptics: Typically chemical agents that are applied to living tissues
• Table 12.1
• Sterilization: The killing or removal of all living cells, viable spores, viruses in a material or on an object
• Sterility: there are no living organisms in or on an object
• Disinfection: The reduction of the number of pathogenic microorganisms to the point that they pose no danger of disease
• X rays and gamma rays are forms of ionizing radiation
• So named because it can dislodge electrons from atoms creating ions
• Damages DNA and produces peroxides (powerful oxidizing agents in cells)
• Deinococcus radiodurans: Able to survive 1000X the amount of radiation that would kill a human (bioremediation of radioactive contaminated sites)
Ionizing Radiation
• If water is heated under pressure, its boiling point is elevated, so temperatures above 100oC can be reached
• Pressure: 15 lbs per square inch (psi)
• 15 – 20 minutes at 121oC
• Prions are highly resistant and must be sterilized longer and at higher temperature (134oC for 18 min)
Autoclaving
Pasteurization
• A process invented by Pasteur to destroy microbes that caused wine to sour, does not achieve sterility
• Kills pathogens: Salmonella and Mycobacterium
• Milk is pasteurized by heating it to 71.6oC for at least 15 seconds (flash method)
• Milk is pasteurized by heating it to 62.9oC for 30 minutes (holding method)
• A definite proportion of the organisms die in a given time interval
• The fewer organisms present, the shorter the time needed to achieve sterility
• Microorganisms differ in their susceptibility to antimicrobial agents
• The most susceptible phase for most organisms is the logarithmic growth phase
The Control of Microbial Growth
• Germicide: An agent capable of killing pathogens and nonpathogens but not necessarily endospores
• Bacteriostatic agent: An agent that inhibits the growth of bacteria
1. Bactericide
2. Bacteriostatic
The Use of Physical Methods in Control of Microbial Growth
• Heat and other physical agents are normally used to control microbial growth and sterilize objects:
1. Heat2. Low Temperatures3. Filtration4. Radiation
Heat• One of the most popular ways to destroy
microbes (flame or boiling)
• Moist or Dry heat may be used
• Exposure to boiling water for 10 minutes is sufficient to destroy vegetative cells and eukaryotic spores
• Steam sterilization (autoclaving) is necessary to destroy the bacterial endospore
Radiation
• Ultraviolet radiation (260nm): is quite lethal in destroying microbes but it does not penetrate glass, dirt films, or water
• Ionizing radiation (IR): An excellent sterilizing agent because of its ability to penetrate deep into objects
• IR will destroy bacterial endospores and both prokaryotic and eukaryotic vegetative cells
Filtration
• Can be used to sterilize substances that are destroyed by heat (drugs, serum, vitamins, sucrose)
• To separate viruses from bacteria (manufacture of vaccines)
• To collect microorganisms from air and water samples (water quality testing)