CHAPTER 3

Sterilization &
Disinfection

Subheading goes here

 IMPORTANT TERMS

• Sterilization: complete killing of all forms of

microorganisms, including bacterial spores
• Disinfection: killing or removing of harmful
vegetative microorganisms.
• Disinfectant: chemical substance used to
achieve disinfection.
• Antiseptic: disinfectant that can be safely
used on living tissues.
 IMPORTANT TERMS
• “-static” - treatments that inhibit rather than kill.
• Eg: refrigeration. (bacteriostatic, fungistatic, etc.).
• Agents or processes which prevent the multiplication / growth of bacteria

• “-cidal” - treatments that kill.

– (bactericidal, fungicidal, viricidal, etc.) (germicidal is a more general).
– Agents or processes by which the bacteria are killed

• Bacteriostatic - Biological or chemical agent that stops

bacteria from reproducing, while not necessarily harming them
otherwise.

• Virucide - An agent (physical or chemical) that kills or destroys

viruses.
 IMPORTANT TERMS
• Aseptic(Asepsis)
– Prevent contamination of person or object by
microbes
• Sanitize
– Removal of pathogens from inanimate objects
– Mechanical or chemical cleaning
– need not sterilize of disinfect
• Contamination
– Presence of living microbes on object
 IMPORTANT TERMS
• Infection
– Presence of living multiplying microbes in
host tissues
– often pathogenic
• Preservation
– Prevention of spoilage
– Control of Contamination
– Bacteriostatic
 SANITIZE vs DISINFECT
SANITIZE
Some types of pathogenic
bacteria can not be killed using the
sanitizers.
30 seconds
agents used for sanitization are
usually used in destroying germs on
surfaces in order to make them
sufficiently safe to be used
DISINFECT
Can kill almost all bacteria.
5 to 10 minutes
normally involves the use of
commercial products, such as heat
or bleach in order to destroy or kill
bacteria found on the surfaces.
The methods chosen depend on…..
 General Rule for Disinfection
• Clean then disinfect
– organic matter inactivates many chemicals
• Use at recommended strength
– Undiluted NOT always best
– 70% alcohol better than 100% alcohol
• Prevent contamination of disinfectant
• Prepare fresh
– deterioration after prolonged storage
 METHODS OF STERILIZATION
• PHYSICAL • CHEMICAL
• Sunlight • Acids & alkalines
• Salts
• Drying
• Halogens
• Dry Heat • Oxidizing agents
• Moist Heat • Reducing agents
• Filtration • Formaldehyde
• Radiation • Phenol
• Soap
• Ultrasonic vibrations
• Dyes
• Aerosol
 PHYSICAL CONTROL
• Heat
1. Advantages - simple, inexpensive, effective penetrates to kill
microbes throughout the object; best method if material being
treated is not damaged by heat.
2. Mode of Action - denatures proteins.

3. Treatments:
  Dry Heat Sterilization
  Moist heat sterilization
  Pasteurization
 Dry Heat Moist Heat Pasteurization
Sterilization
EX: flaming loops, Ex: boiling or in Standard treatment:
tubes in lab & hot air autoclaves heat to 63oC for 30
ovens (171oC, 1hr., min. or 72oC for 15 sec.
160oC for 2 hr., 121oC
for 16 hrs.)
used to sterilize effective at a lower causes minimal
materials that can temperature than dry damage to the
withstand high temps. heat & it penetrates product
more quickly used to slow spoilage
of milk & dairy
products, wine, beer
any materials does not kill limits growth, but does
damaged by moisture. thermophiles, not sterilize
endospores;
 ULTRAVIOLET (UV) LIGHT
• UV lamps useful for chemical-free
disinfection of air and water and also
possibly for decontamination of
environmental surfaces
• Broad-spectrum microbicidal action
• Require regular cleaning and periodic
replacement
 Filtration
• Removal of microbes from air or
heat-sensitive liquids
• Disinfectant-impregnated filters
may inactivate trapped
microorganisms
• Example: High-efficiency
particulate air (HEPA) filters
• All filters must be checked for
integrity and replaced as
necessary
 PHYSICAL CONTROL
• Cold
  Effect - microbiostatic; does not sterilize; slows
down enzymes

Refrigeration Freezing
- preserves food because it - kills most bacteria, but
stops the growth of most survivors can remain alive for
species of microbes (slows long periods in the frozen
chemical reactions). state
- most disease-causing - bacteria cultures can be
microbes are mesophiles, not preserved by rapid freezing.
psychrophiles; an exception is
Listeria spp., which causes
listeriosis (food poisoning
 Chemical agents
• Wide variety in susceptibility
• Growing cells more susceptible than
resting cells or spores
• Gram +ve more susceptible than Gram -ve
• Mycobacterium (TB) more resistant
• Hepatitis virus very resistant
 CHEMICAL CONTROL
• The effectiveness of a chemical
antimicrobial agent is affected by time,
temperature, pH, and concentration
 CHEMICAL CONTROL
• Types of Germicides Surfactants
  Structure - compounds with hydrophilic & hydrophobic
parts.
  Mode of action - Penetrate oily substances in water & break
them apart into small droplets that become coated with
surfactant molecules. The hydrophobic end of the surfactant stick
into the droplets & the hydrophilic end is attracted to the water.
The result is an emulsion, a fine suspension of oily droplets in water,
which can now be rinsed away.
 CHEMICAL CONTROL
• Effect of soaps  Wetting agents
& detergents - are surfactants
wash away that are often
microbes, but used with other
chemical agents
do not kill
to help the agent
them. penetrate fatty
substances.
Surfactants are
not germicidal by
themselves!
 CHEMICAL CONTROL
• Alcohols
  Structure - compounds with a hydroxyl
group (-OH).
  Mode of Action – when mixed with water
disrupt lipids in cell membranes & denature
proteins.
 CHEMICAL CONTROL
• Ethanol & Isopropanol - widely used as skin
antiseptics; a 50 to 70% solution in water is the most
effective concentration (one of the few exceptions
to the rule: increase effectiveness by increasing
concentration); does not sterilize skin because it
evaporates quickly and does not penetrate deeply
enough into skin pores. (kills Mycobacterium)
  Main disadvantage - do not kill endospores.
 Halogens(Iodine)
• Applications
– Skin antiseptic
– Environmental disinfectant
• Mode of Action
– Iodination of Tyrosine
• Prepare fresh
– Dark brown - active
– Straw Yellow - inactive
• Clean area first
– organic matter reduces activity
 Halogens(Chlorine)
• Bacteriocide, fungicidal, virucidal
• Applications
– disinfection of water, sewage etc
• Mode of Action
– Oxidizes proteins
• Features
– corrosive
– smells
 Halogens(Chlorine)
• Chlorine gas • Inorganic chlorine
– Poisonous – Bleach NaOCl
– forms HOCl – hypochlorite Ca(0Cl)2
(hypochlorous acid) in – chloramine
water
• effective against
• Applications – Hepatitis
– water supplies – HIV
– swimming pools
– sewage effluent
 Metal Ions
• Silver
– 1% AgNO3 eye drops
– gonnorrheal neaonatal opthalmitis
• Mercury
– Hg Cl2 Skin antiseptic
– Mercurochrome
• Copper, zinc
– fungicide, algacide
– pressure treated wood
Kill curves of various antiseptics
Soap & water

Tincture of Merthiolate
% survivors

Aqueous Zephiran

Tincture of Zephiran
Tincture of
Iodine

70% ethanol
Time (sec)
 Activity time…
• In a pairing group, choose one of the physical & chemical
control that not discussed yet.
• Discuss about how the physical & chemical control works,
examples, advantage and disadvantage
• Don’t forget to present