Sterilization &

Disinfection

Dr.Abid Hussain Chang

 Sterilization;
Killing or removal of all microorganisms including
the resistant bacterial spores
 Disinfection;
Freeing of an article from harmful
microorganisms, the spores usually survive.
 Antiseptics;
Chemicals used to kill microorganisms on skin
surfaces & mucous membranes.
 Methods;
 1-Physical;
Heat
Ionizing radiation
Filtration
 2-Chemical
 Radiation;
2 types
1.Ultraviolet light U/V; 240-280 nm is most
effective
.Lesser penetrating power so are less
effective
.Spores are resistant to it & require a higher dose
.Can damage skin, cornea
.Therefore use limited esp for operating rooms
when not in use
 2.X-rays;
. Have higher energy & penetrating power
than U/V
.Kill by production of free radicals
.Vegetative cells are killed but not spores
.Used for heat sensitive objects
sutures, surgical gloves, plastic items
 Filtration;
.Works mainly by physically trapping particles
larger than pore size
.Useful for making certain solutions & fluids
bacteria free, which are heat sensitive,
e.g antisera & toxins
Methods;
 1-Physical;
Heating;
A very effective, reliable, widely accepted
method.
Generally, it kills by denaturing proteins.
 1-Physical;
Heating;
Applied in two ways
Dry heat
Moist heat
 Dry Heat;
Less efficient than moist heat
.Applied as,
Red Heat; put directly in flame till red hot
inoculating wireloops
tips of needles & forceps
Flaming; direct exposure for a few seconds, it
may be useful for scalpels & neck of flasks
 Hot air oven;
applied for substances which can withstand
high temperatures (160-180c)
.Glassware
.Throat swabs
.Syringes
.Powders, fats, oils, greases, impermeable to
moist heat
160c for 1 hr is reqd
Filtration;

 Since long filters have been used in one form

or other to purify water & sewage
.It is preferred method to sterilise certain
solutions
Membrane filters;
Used widely nowadays
Consist of cellulose ester
Available in pore size of .05-12 micron
Required pore size for filtration is 0.2-0.22
micron
.This is useful for
thermolabile parenteral & ophthalmic
solutions
.Sera sterilization
.Clarification of water supplies
 Moist Heat;
Includes;
Pasteurization; (< 100c)
used for milk
Temp is 63-66c for 30 min or
72c for 20 sec.
Boiling; (100c)
.Contineously for 90 min-culture media
.Intermittently for 20-30 min for 3
consecutive days (Tyndallization)
.for sugar media which are
destroyed by prolonged heating
 Temperature > 100c ( Autoclaving);
.Most effective method of sterilization

.Destruction of bacterial spores needs >100c

which is possible only with Autoclave
.Autoclave provides moist heat (steam) at a pressure
of 15 lb/in2 reaching 121c for 15-20 m
Autoclave
 Consists of a vertical or horizontal cylinder of
gun metal or stainless steel in a supporting
iron case
 Screw clamps in lid
 Airtight by washer
 Disharge tap for air & steam, pressure guage, a
safety valve on lid
 Heating by electricity/gas
 Principle;
.Water boils at 100c,vapour pressure equals
760mm Hg or 14.7 lb/n2.
.If water heated within closed vessel, pressure
increases with corresponding temp rise
 Uses;
To sterilize surgical instruments,
linen
bacterioloical culture media
 Chemical;
1-Disruption of cell membranes;
Alcohol;
Ethanol with water (70%) is more efficient
than pure (100% ) to clean skin before venipuncture
 Detergents;
Benzalkonium chloride used as skin antiseptic
 Phenols;1st disinfectant used in OT
Lysol, phenol, cresol, dettol are used for
disinfection of discharges, floors, bathrooms
 2-Modification of proteins;
.Chlorine; for water purification
Treatment of swimmng pools
Included in hypochlorite
(disinfectant)
.Heavy metals;
mercury & silver used as antisepics
 Silver nitrate drops prevent gonococcal ophthalmia
neonatorum
 Silver sulfadiazine prevents burn wound infections
 Iodine;
.Most effective antiseptic
Tinc iodine for cleaning skin before blood culture
.Iodophors (iodine+detergents) used to prepare skin
before surgery
 Hydrogen peroxide;
Used to clean wounds & contact lenses
Limited against organisms producing catalase
that degrades it
 Formaldehyde &
Glutaraldehyde;
To sterilize respiratory therapy equipments
 Ethylene oxide;
For heat unstable surgical instruments, plastics
 Acids & Alkalis;
Effective against most
Mycobacteria are resistant to 2% NaOH used
to liquefy sputum before culture
3-Alteration of Nucleic Acids;
.Crystal violet (gention violet)
skin antiseptic
.Malachite green is included in Lowenstein
Jensen medium (for growth of
M.tuberculosis).
Inhibits growth of unwanted organisms