Sterilization and

Disinfections
 Sterilization
Freeing of an article from all living
microorganisms includes bacteria and
their spores, fungi, parasites and viruses.
Sterilization is an absolute term means
germ free objects.
Sterilization occurs by:
 Physical methods.

Chemical methods.
 Physical methods
Heat:
 Exposure of the objects to heat will kills microbes by
coagulation of protein, denaturation of enzymes and
oxidation.
Filtration:
 Sterilization through removing of microbes from fluids
by exposing to small size filter. Used for heat
sensitive fluids like serum, CHO and urea.
Radiation:
 Exposure to irradiation causes denaturation of
proteins and enzymes.
 Sterilization
by
Physical
methods

Heat Radiation Filteration

 sterilization by heat

Heat

Dry heat Moist heat

 Dry heat

Dry heat

Hot air
Red hot Flaming Incineration
oven
 Red hot
Exposure of wires and forceps to the
Bunsen flame until it becomes red hot,
then cool down and use.
Used for wire loop, straight wire, forceps,
scalpels and metal rods.
 Flaming
Slowly passing of an objects to the
Bunsen flame will reduce the number of
microorganisms.
The Bunsen flame give partially sterile
area around it in which used for work
(preparation of smears, cultivation,
subculture, etc….).
Used for sterilization of the mouth of
bottle, flasks, containers and test tubes.
Flaming
 Hot air oven
Instruments consist of heater,
thermometer, thermostat, perforated
shelves, door.
Kills microbes by oxidation-reduction.
Thermal death point and Thermal death
time:
 160C for 60 min.
 170C for 40 min.
 180C for 30 min.
 Check the efficiency
BoweiDick test (Adhesive tab).
Browne’s tube No 3. (Red ----- green).
 Incineration
Is treating of an objects to heating over
250 until become black.
Done for used equipment.
 Moist heat

Moist heat

Less than 100C At 100C Above 100C

 Less than 100C
Pasteurization of milk:
 Holding method (65C for 30 min)
 Flash method (72C for 20 sec)
Inspeciation:
 Heating at 80C for 30 - 1hrs until coagulation
of protein.
 Used for preparation of dorset egg medium
and L.J medium.
Preparation of vaccine:
 By heating at 56C for 30-60 min.
 At 100 C
Steaming (Koch steamer):
 Single exposure of the microbe to steam at
100C for 90 min.
 Tyndalization:
Steaming at 100C for 30 min for 3 successive
days.
 1st day kill vegetative bacteria and germinate sporulated
one ---- Put on the bench.

2nd day kills all vegetative bacteria ---- Put on the bench.

3rd day insure complete sterilization.

Poilling:
 At 100C for 30 min.
 Above 100C (Autoclaving)
Depends on steam and pressure.

Steam is a hot sticky air able to penetrate through
things.

Pressure will rise the temperature from 100C to 121C
Kills microbes and their spores by coagulation of
protein and denaturation of enzymes.
Make complete killing of bacteria, their spores,
fungi and their spores, parasites and viruses
including Envelop and non Envelop virus.
Thermal death point and thermal death time:
 121C (15 bound or 1.1 bar) for 15 min.
 Flash autoclaving at 134C for 4-5min.
 Radiation

Sterilization by radiation kills microbes by causing
mutation to the cellular protein and disrupting cellular
elements.
 Used for plastic syringes, disposable plastic and dental
e
2 types:

Ionizing radiation:
Depends on ionization of water which forms highly reactive
hydroxyl group react with cellular components specially DNA
causing mutation.
E.g.: X-rays, Gamma radiation and high energy electron
beams.
Have a wavelength less than 1 nm.

Non ionizing radiation:
using short wave length rays like Ultraviolet with a wavelength
of 260 nm.
Causing mutation by making thymine dimers.
Used for sterilization of plastic and surgical room
 Filtration
Sterilization by mechanical removal of
pathogenic microbes by passing through
membrane filter.
Used for sterilization of heat sensitive fluids like
serum, glucose, urea, and Amino acids.
Different filtration units is used according to the
amount of filtrate and pores of the filter:
 Seitz filter ---- use asbestos
 Chamberland filter ------ use ceramic
 Sintered filter ----- use glass filter.
 Milipore or membrane filter ---- use filter paper.
 Disinfection
Chemical methods of sterilization
Disinfection:
 Is removing of pathogenic microorganism or
reducing their number on the exposed area.
 Unable to destroy spores and some could not
kill non envelop viruses.
Factors affects disinfection action:
 Type of disinfection used.
 Concentration of disinfectant.
 Type of microorganism.
 Number of microorganism.
 Time of exposure.

Temperature.

Presence of organic compound.
1. Phenolic group of disinfectant:
E.g.: Phenol crystal, Dittol, Lysol, Cresol.
Injuring lipid containing plasma membrane leads
to leakage of cellular contents.
Remain active in the presence of organic
materials, stable and persist for long period of
time.
Suitable for disinfecting pus, blood and sputum.
Active against G+ve, G-ve, Mycobacterium &
viruses.
 2. Bisphenol:
Contains 2 groups of phenol.
Hexaclorophenol used for surgical and
microbial control, Excessive use for infant
could leads to neurologic damage.
Triclosan found in antimicrobial soaps,
inhibits an enzyme needed for synthesis of
fatty acid which affects the integrity of
plasma membrane.
 3. Biguanides:
E.g.: Chlorohexidine.
Broad spectrum activity as disinfectant of
skin and mucus membrane due to it is
ability to bind to mucus membrane.
Cause injury to plasma membrane.
 4. Haloges:
Iodine have broad spectrum activity to many
bacteria, spores, fungi and some viruses, it
binds to certain amino acids of enzyme and
proteins.
Iodophore (Betadine and Isodine) is non
pigmented iodine comprise from iodine + organic
materials to release the iodine slowly. Used for
skin disinfection and wounds.
Chlorine in shape of gas or solution combined
with water to give HOCl (Oxidizing agents).
Used for swimming pools, drinking water and
sewage.
 5. Alcohols:
Causes denaturation of proteins, disrupt
membrane and dissolve lipids.
They able to act and Evaporate (Volatile).
Used as skin disinfectant and in vein
puncture in a concentration of 70%.
 6. Heavy metals:
Silver, mercury and copper
Causes denaturation of proteins when bind to it.
Silver nitrate is used as a droper to avoid
opthalmia neontam by N. gonorrhoea.
Silver sulfadiazine (Flamazine) is used in
treating infection associated with burns.
Copper sulfate is used to destroy green algae
(effective in one part/million of water).
 7. Dyes:
Gention violet, Crystal violet and Eosine
are very effective antiseptic.
 8. Surface active agents:
Soap and other detergent make
mechanical remove of microbe by
scrubbing of dead tissue so reduce their
number.
Quaternary ammonium compound are
positively charged molecules kills G+ve
bacteria. Affects plasma membrane and
changes cell permiability. E.g.: Citerimide.
 9. Aldehyde:
E.g.: Formaldehyde and Gultraldehyde.
Inactivate proteins by covalent cross-links with
COOH, NH, OH and SH.
Used for disinfect hospital instruments, benches
and room. E.g.: Formaldehyde and
Gultraldehyde.
Inactivate proteins by covalent cross-links with
COOH, NH, OH and SH.
Used for disinfect hospital instruments, benches
and room.
 10. Ethylene oxide gas:
Used to sterilize plastics in closed
chamber similar to autoclave.
Denaturate proteins.
Bacteriocidal, fungicidal and kills viruses.
Used at high temperature 60C for 4-24
hrs.
Carcinogenic.
 11. Peroxygens:
H2O2 is an oxidizing agents kills many
microbes especially who fail to produce
catalase enzymes (Anaerobic bacteria).
Used as disinfectant for gas gangrene.
Ozone (O3) also toxic for microbes and
used instead of chlorine in treating
drinking water.