02-Aug-18

Sterilization and Disinfection

Definitions
• Sterilization: complete killing, or removal, of all organism
from a particular location or material
• Pasteurization: the use of heat at a temperature sufficient
to inactive important pathogenic organism in liquids
• Disinfection: the destruction of pathogenic microorganisms
by processes that fail to meet the criteria for sterilization
• Antiseptic: disinfectant agents that can be used on body
surfaces
• Sanitization: a less precise term with meaning somewhere
between disinfection and cleanliness. (in house-keeping and
food preparations contexts)
• Asepsis: processes designed to prevent microorganisms
from reaching a protected environment.

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Definitions
• Bactericide: agent to destroy bacteria
• Bacteriostatic: substance that prevent
bacterial growth
• Disinfectant (germicide): agent that kills
micro-organisms capable of producing an
infection.
• Decontamination: get rid of contamination
some of the microorganisms.

DEFINITIONS
• Sterilization: total inactivation of all forms of
microbial life → ability to reproduce
• Disinfectant (germicide): agent that kills micro-
organisms capable of producing an infection.
• Antiseptic: opposes sepsis or putrefaction by killing
bacteria or preventing their growth. (on living
tissue)
• Decontamination: get rid of contamination (some of
the microorganisms.

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Sterilization versus Disinfection

• Sterilisation reduction in microorganisms by one million

(106 or more than 99.9999% are killed)
• High Level Disinfection - all microorganisms, with the
exception of small numbers of bacterial spores are killed.
• Intermediate Level Disinfection - Mycobacterium
tuberculosis, most viruses and fungi are killed, but not
necessarily bacterial spores.
• Low Level Disinfection - most bacteria, some viruses and
some fungi are killed. Complete absence of resistant
microorganisms e.g. tubercle bacilli or bacterial spores
cannot be relied on.

Disinfectant vs. Antiseptic

• Disinfectant
– antimicrobial agent used on inanimate objects

• Antiseptic
– antimicrobial agent used on living tissue

• May exert cidal or static effect

• cidal vs. static
– Bactericidal - kills bacteria
– Bacteristatic - inhibits bacterial growth

– Fungicidal vs Fungistatic
– Virucidal vs Virustatic

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Method of Disinfection and Sterilization

DEATH RATE OF MICROORGANISMS

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Physical Methods used to control Microbial

Growth
• 1. Heat
– Dry Heat
– Moist Heat
• 2. Filtration
• 3. Radiation
– Ionizing Radiation
– Non-Ionizing Radiation
• 4. Pasteurization (Heat)

Dry Heat
• 1. Direct Flaming
– Inoculating Loop and Needle 100% effective

• 2. Incineration
– disposable wastes (paper cups, bags, dressings)

• 3. Hot Air Sterilization

– Oven ( 170 C for 2 hours)
– mainly use for metal and glass
• gauzes, dressings or powders

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Moist Heat
• 1. Boiling Water
– Maximum temperature +/- 100C
– kills vegetative bacterial cells, Fungi and many viruses
– not effective for endospores and some viruses
– Hepatitis (20 min)
– Some spores may survive boiling water for up to 20 hrs
– Require minimum equipment but cumbersome

Moist Heat
2. Autoclave (Steam under pressure)

• Preferred method of sterilization

• Moist heat (steam) at increased pressure for a defined time.
• Specifically designed for porous loads (comprising all wrapped
articles), unwrapped instruments and utensil, or aqueous
liquids.
• Can be used for most items (e.g. surgical instruments, fabrics,
etc.).
• Ability to kill spores should be checked weekly.

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Autoclave

Filtration
• Sterilization: heat-labile
materials (lab)
• Mechanisms:
- Electrostatic & absorption
phenomena
- Physical construction of filters
• Types:
- Old types : Berkefeld,
Chamberland, Seitz
- Membrane filters : porous disks
of cellulose

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Filtration
• Removes microorganisms from heat-labile
fluids

– culture media
– enzymes
– vaccines
– antibiotics

Membrane filters
- Pore size: 14-0.023 µm.
- Most widely used: 0.023 µm.
- Practical used: serum, plasma, etc.
- Two-dimensional screen.
- Mechanisms:
a. Particles > pore : retained on filter surface
b. Particle < pore: retained by van der Waals
forces.

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Filter

Radiation
• 1. Ionizing Radiation

– gamma rays & x-rays

• penetrates most substances

– Used on substances that could be damaged by heat

• plastic petri dishes
• plastic syringes
• catheters
• surgical gloves

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Radiation
• 2. Non-Ionizing Radiation
– UV Light
• does not penetrate plastic, glass or proteinaceous
matter

– Used to reduce microbial populations

• hospital rooms
• nurseries
• operating rooms

Laminar Flow

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Pasteurization
• Disinfection - not sterilization (removes unwanted
organisms)
• Mycobacterium tuberculosis
– 63 C for 30 minutes
– 72 C for 15 seconds (HTST)

Gas
• Used for objects damaged by heat or radiation.
• Requires aeration step after sterilization

CHEMICAL AGENTS
Factors Affecting Disinfectant Potency
1. Concentration (Cnt = K)
2. Time of Exposure
3. pH
4. Temperature
5. Nature of the Organisms
6. Presence of Extraneous materials → organic
matter: serum, blood, pus, etc.

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Chemical Methods used to control Microbial

Growth
Use of chemical agent
“High Level Disinfectants”
• Substances able to kill spores, tubercle bacilli, and
viruses given enough time.
– Only acceptable alternative when heat-labile and absence
of other low temperature sterilization process
– Examples
• Glutaraldehyde
• Formaldehyde

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