Sterilisation

Dr.B.Reena Rajkumari
 Controlling Microorganisms
• General Considerations in Microbial Control
– Sterilization: the destruction of all microbial life
– Disinfection: destroys most microbial life,
reducing contamination on inanimate surfaces
– Antisepsis: destroys most microbial life, reducing
contamination on a living surface
– Decontamination: the mechanical removal of
most microbes from an animate or inanimate
surface

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Terminology and Methods of Microbial
Control
• Sterilization
– Removes all viable microorganisms including
viruses
– Material is said to be sterile
– Usually reserved for inanimate objects
– Mostly performed with heat
– Sometimes chemicals called sterilants are used

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 Disinfection
• The use of a physical process or chemical
agent (disinfectant) to destroy vegetative
pathogens
• Does not destroy bacterial endospores
• Usually used only on inanimate objects
• Also removes toxins
• 5% bleach solution

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 Antisepsis
• Antiseptics applied directly to exposed body
surfaces to destroy or inhibit vegetative
pathogens
• Sepsis: the growth of microorganisms in the
blood and other tissues
• Asepsis: any practice that prevents the entry
of infectious agents into sterile tissues

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 Decontamination
• Used when actual sterilization isn’t needed but need to
decrease the risk of infection or spoilage (ex. food
industry)
• Sanitization: any cleansing technique that
mechanically removes microorganisms to reduce
contamination to safe levels
• Sanitizer: compound such as soap or detergent that
sanitizes
• Sanitary: may not be free from microbes but are safe
for normal use
• Degermation: reduces the numbers of microbes on
the human skin (ex. alcohol wipes)

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 The Agents Versus the Processes
• –cide: to kill
– Bactericide: chemical that destroys bacteria (not endospores)
– Fungicide: a chemical that can kill fungal spores, hyphae, and
yeasts
– Virucide: a chemical that inactivates viruses
– Sporicide: can destroy bacterial endospores
– Germicide and microbicide: chemical agents that kill
microorganisms
• Stasis and static: to stand still
– Bacteristatic: prevent the growth of bacteria
– Fungistatic: inhibit fungal growth
– Microbistatic: materials used to control microorganisms in the
body, for example

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Relative Resistance of Microbial Forms
• Primary targets of microbial control: microorganisms
that can cause infection or spoilage that are constantly
present in the external environment
• Contaminants that need to be controlled
– Bacterial vegetative cells and endospores (so resistant, the
goal is sterilization)
– Fungal hyphae and spores
– Yeasts
– Protozoan trophozoites and cysts
– Worms
– Viruses
– Prions

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Practical Concerns in the Use of Heat:
Thermal Death Measurements
• Temperature and length of exposure must be
considered
• Higher temperatures generally allow shorter
exposure times; lower temperatures generally
require longer exposure times
• Thermal death time (TDT): the shortest length of
time required to kill all test microbes at a
specified temperature
• Thermal death point (TDP): the lowest
temperature required to kill all microbes in a
sample in 10 minutes
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 Using Heat to Destroy Microorganisms and
Viruses

• Heat treatment useful for microbial control

– Reliable, safe, relatively fast, inexpensive, non-toxic
– Can be used to sterilize or disinfect
– Methods include moist heat, dry heat
• Moist heat: irreversibly denatures proteins
– Boiling destroys most microorganisms and viruses
• Does not sterilize: endospores can survive

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Classification of Moist Heat Methods

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• Tyndallisation -100°C for 20 minutes – 3days
– First day – Vegetative Bacteria are killed
– Second day – endospores -> bacteria are killed
– Third day – Complete sterility of media is ensured
• Inspissation - 80°C for 30 minutes – 3days (similar
to Tyndallisation)

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 Pasteurization
• Used to disinfect beverages
• Heat is applied to liquids to kill potential agents of infection and
spoilage, while retaining the liquid’s flavor and food value
• Special heat exchangers
– Flash method: expose to 71.6°C for 15 seconds
– Batch method: expose to 63°C to 66°C for 30 minutes
– High-temperature–short-time (HTST): most products
– Milk: 72°C for 15 s; ice cream: 82°C for 20 s
– Ultra-high-temperature (UHT): shelf-stable boxed juice and milk;
known as “ultra-pasteurization”
– Milk: 140°C for a few seconds, then rapidly cooled
• Does not kill endospores or thermoduric microbes

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 Autoclave

• Moist heat
denatures
proteins
• Autoclave:
Steam under
pressure

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Figure
Autoclave - Vertical

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Steam Jacketed Horizontal autoclave

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Biological Indicator

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Time temperature chart

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Hot air oven

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Hot air oven

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Laminar Air Flow

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