‫بسم هللا الرحمن الرحيم‬

‫اللهم صلى وسلم وبارك على سيدنا محمد‬

 By
Prof . Eman El-Seidi
Medical Microbiology & Immunology Department,
Faculty of Medicine, Cairo University
 Decontamination

• Reduction of pathogenic microorganisms to a

level at which items are safe to handle.

• Decontamination includes sterilization and

disinfection processes.
 Cleaning
 Removal of foreign material (organic or
inorganic contaminants) from medical
devices as part of decontamination process.

 Usually done with water & soap, detergents

or enzymatic products.

 Meticulous cleaning must always precede

disinfection or sterilization.
 Sterilization
 Definition: Validated process
used to render a product free of all
forms of viable microorganisms
including all bacterial spores.

 Main sterilization processes:

1. Steam under pressure
2. Hydrogen peroxide gas plasma
3. Ethylene oxide gas
4. Dry heat
 Sterilization
 Uses:
1. Culture media preparation.

2. Critical items intended to enter

the vascular system & sterile
tissues such as vascular
catheters & surgical instruments.
 Disinfection
 A process that eliminates most, if not
all, pathogenic microorganisms except
spores.
 Uses: devices or equipment that do not
penetrate tissues but come in contact
with:

1. Skin (e.g., stethoscope diaphragm

swabbed with 70% alcohol)

or

2. Mucous membranes (e.g., immersion

of endoscope in 2% ortho-
phthalaldehyde (OPA) disinfectant for
12 minutes)
 Definitions
 Disinfectant: usually a chemical agent
(but sometimes a physical agent) applied
to inanimate objects that achieves
disinfection (e.g., phenolics).

 Antiseptic: a chemical disinfectant

which can be safely applied to skin &
mucous membranes but not suitable for
systemic administration (e.g., 70% alcohol
to prepare skin for injection, preoperative
skin preparation with alcohol-based iodine
compound in surgical operations)
 Definitions
 Germicide: Agent that destroys
microorganisms; may be virucide,
bactericide, fungicide, sporicide or
tuberculocide.
 The term germicide includes both antiseptics &
disinfectants:
 Antiseptics → applied to living tissue and skin.
 Disinfectants → applied only to inanimate
objects.

 Sterilant: chemical germicide that

achieves sterilization.
Disinfection Levels
 High Level Disinfectants

 Germicides that kill all microbial

pathogens except large numbers of
bacterial spores.

 HLD kill Mycobacterium spp.

 Examples:
 OPA for endoscopes

 Hydrogen peroxide for contact lens

 Chlorine for blood spills

Intermediate Level Disinfectants

 Germicide that kills all microbial pathogens

except bacterial spores.

 ILD might be cidal for Mycobacterium spp.

 Examples: isopropyl alcohol & iodophors.

 Low Level Disinfectants

 Germicide that kills most vegetative

bacteria (except tubercle bacilli) and
lipid-enveloped and medium-sized
viruses (e.g., HIV & HBV).

 e.g., quaternary ammonium

compounds for disinfection of floors
and food preparation areas.
 ? Resistant

Sterilization
(Steam & Plasma)

High LD
(Glut, OPA & H2O2)

Intermediate LD
(Alcohols & Chlorines)

Low LD
(Alcohols &
Chlorines)

Susceptible
 Main Methods of Disinfection

1. Chemical disinfectants (see above).

2. Boiling water: Boiling (100°C) for 20 minutes achieves
high level disinfection → useful in emergencies, if sterilizer is
not available.

3. Pasteurization of milk: by heating at 63°C for 30 min.

or at 72°C for 20 sec., followed by rapid cooling (<10oC)
destroys important pathogens e.g. M. tuberculosis, Brucella,
Salmonella and Coxiella burnetti.
 Main Methods of Disinfection

4. Thermal disinfection (by hot water) in special

washing machines (e.g. for linen in hospital laundry,
dishes and devices which cannot withstand higher
temperature).

5. Ultraviolet radiation (UV): have weak penetration

power used only for air (in O.R.) & surface
disinfection (e.g., laboratory safety cabinets).
Main methods of Sterilization
Steam Sterilization

 Most safe & commonly used

sterilization method
 Steam Sterilization Principle
• Accomplished in an autoclave.
• The sterilizing agent: moist heat (saturated steam)
under pressure for a specified exposure time & at a
specified temperature.

• Parameters of steam sterilization:

1. Saturated steam,
2. Pressure,
3. Temperature,
4. Exposure Time
Steam Sterilization Principles
• Two common temperatures: 121oC & 134oC.

• Minimum exposure periods for sterilization are:

– 20-30 minutes at 121oC in a gravity displacement autoclave or
– 3-6 minutes at 132oC–135°C in a prevacuum autoclave.

• Mode of action: moist heat destroys microorganisms

by coagulation & denaturation of enzymes & structural
proteins.
Types of Steam Sterilization
Gravity displacement
autoclave

Prevacuum
autoclave
 Steam Sterilization
Advantages

1. Nontoxic.

2. Inexpensive.

3. Rapidly heats & penetrates fabrics.

4. The most reliable.

Monitors of Steam Sterilizers
(Autoclaves)
 Mechanical Indicators

• A printout or graph
that monitors the
time, temperature
& pressure of the
sterilization cycle.
 Chemical Indicators
(or Integrators)
• Chemically impregnated paper strips that must be used
with each sterilization cycle to monitor the temperature
or time & temperature.
• Visible colour changes occur.
Biological Indicators
1. Paper strips impregnated with

spores of G. stearothermophilus.

1. They are placed at the coldest point

of the chamber.

1. After finishing the cycle of sterilization, they are

incubated in a fluid medium at 37°c for 48h.

2. Absence of bacterial growth indicates an efficient

sterilization cycle.
 Biological
Indicators
Sterilization cannot be
proved except by culturing
 Hydrogen Peroxide
Gas Plasma
• Gas plasmas generated in an
enclosed chamber under deep
vacuum using radio frequency or
microwave energy to excite the gas
molecules & produce charged
particles, many of which are in the
form of free radicals.
Radio-frequency energy
generates plasma from
• Free radicals interact with essential vaporized H2O2
cell components (e.g., enzymes,
nucleic acids) → disrupt the
metabolism of microorganisms, in
addition to the direct inactivation by
hydrogen peroxide.
 Plasma
• Plasma is a partially ionized gas, in which a certain proportion of
electrons are free rather than being bound to an atom or
molecule.
• The ability of the positive & negative charges to move
independently makes the plasma electrically conductive so that
it responds strongly to electromagnetic fields
• Plasma therefore has properties quite unlike those of solids,
liquids, or gases and is considered to be a distinct state of
matter. Plasma typically takes the form of neutral gas-like
clouds, as seen, for example, in the case of stars. Like gas,
plasma does not have a definite shape or a definite volume
unless enclosed in a container. Unlike gas, in the influence of a
magnetic field, it may form structures such as filaments, beams
and double layers
Plasma Lamp
 Hydrogen Peroxide
Gas Plasma
• Time of sterilization cycle about
50 min.
• Uses: Medical materials & devices that
cannot tolerate high temperatures &
humidity (plastics, electrical devices &
corrosion-susceptible metal alloys).

• Biological indicator:

G. stearothermophilus spores.
 Ethylene Oxide Gas Sterilization
• Time of sterilization cycle: from
3 - 6 hours.
• Expensive & toxic.

• Uses: instruments that cannot

be subjected to steam.
• Biological indicator: Bacillus
atrophaeus spores.
Peracetic Acid Sterilization

• Used to sterilize medical, surgical & dental

instruments (e.g., endoscopes,
arthroscopes).

• Peracetic acid denatures proteins, disrupts

cell wall & oxidizes microbial proteins &
enzymes.
 Dry Heat Sterilization
1. Incineration: dead animal bodies, infectious hospital waste such as
used surgical dressings, needles….etc.

2. Red heat: Inoculating wires, loops and points of forceps are

sterilized by holding them in the flame until they are red hot.

3. Hot air ovens (Dry Heat Sterilizers).

 Hot Air Oven
(Dry Heat Sterilizer)
• Sterilizing agent: dry hot air.
• The most common time-temperature relationships are:
1. 170oC for 60 minutes,
2. 160oC for 120 minutes
3. 150oC for 150 minutes.

• Biological indicator: Bacillus atrophaeus spores.

• Mode of action: killing by oxidation of microbial cell
constituents.
• Uses: materials that might be damaged by moist heat
(e.g., glassware, powders, oils, sharp instruments).
 Hot Air Oven
Dry Heat Sterilizer
• Advantages :
– nontoxic

– relatively inexpensive

– Non-corrosive for metal and sharp instruments

• Disadvantages:
– slow rate of heat penetration

– time-consuming

– high temperatures not suitable for most materials

 Ionizing Radiation

• Obtained by cobalt 60 gamma rays or electron

accelerators (β-rays).
• Has a high penetrating power.
• Uses: sterilization of prepacked heat-sensitive items
such as bone grafts, surgical sutures, disposable
plastic syringes, gloves, catheters & plastic petri
dishes.
• Biological indicator: Bacillus pumilus spores.
 Filtration
• A process used to remove bacteria from thermolabile fluids
or air that cannot be purified by any other means.

• Thermolabile fluids (antibiotic solutions, hormones,

vitamins).

• Fluids passed through bacterial membrane filters with pore

size as small as 0.22 µm.

• Air supplied to critical areas (O.R., drug factories &

laboratory biosafety cabinets) passed through HEPA filters.
 Filtration
• The endopigment producing Serratia marcescens
may be used to test the efficiency of bacterial
membrane filters.
•

• Spores of the fungus Aspergillus may be used to

test the efficiency of HEPA filters.
 Bacterial
Membrane Filters
HEPA Filter
 Microwaves

• Microwaves are used in medicine for disinfection of soft

contact lenses, dental instruments, dentures, milk &
urinary catheters.

• The microwaves produce friction of water molecules in

an alternating electrical field → generation of lethal heat.

• Process has not been validated.

 Ozone (O3)
• Used as a sterilant & drinking water disinfectant.

• Consists of O2 with a loosely bonded third oxygen

atom that makes ozone a powerful oxidant that
destroys microorganisms.
 Formaldehyde Steam
• Low-temperature sterilization method that involves use of
formalin, which is vaporized into a formaldehyde gas
admitted into the sterilization chamber.

• Uses: heat-sensitive medical equipment such as mechanical

ventilator & incubators for neonates.

• Unfortunately formaldehyde is a mutagen & a potential

human carcinogen → must be regulated to guarantee
permissible exposure limit of HCWs for formaldehyde.
 Others

• Vaporized hydrogen peroxide.

• Vaporized peracetic acid.

• Gaseous chlorine dioxide.

• Infrared radiation.