CONTROL OF

MICROBIAL GROWTH
Physical and Chemical Method of Sterilization
Prepared: Aida V. Garcia, MAN, RN
MEDICAL ASEPSISSURGICAL ASEPSIS
• Or sterile technique
• Or clean technique
• Procedures aimed in eliminating
• Involves procedures / m.o. From the area of the
practices that reduce the surgical site & OR area
number & prevent
spread in gen. Clinical
setting

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 OBJECTIVE

• Compare and contrast the methods used

to control microbial growth in daily life and
hospital setting.

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 STERILIZATION

PHYSICAL CHEMICAL
METHODS METHODS

• 1. Heat
Depends in
• 2. Radiation circumstances
• 3. Filtration & degree of
• 4. Mechanical protection
removal (washing)
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PHYSICAL METHOD

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PHYSICAL METHOD: HEAT

• simple, cheap & effective method of killing

pathogens.

• 1) moist heat
• 2) dry heat
• 3) radiation
• 4) high pressure
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•heat is absorbed by exterior surface of
item/then passed inward to next layer until
entire item reaches the temp. needed.

DRY HEAT
DESTROYS M.O. BY CAUSING
COAGULATION OF PROTEIN / DRY
 DRY HEAT

• Open flame
• Hot air oven
• incineration

Tubes, oils, jellies, powders

glassware (test tube / petri dish)
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Blue flame – sterilize…
scalpel, mouth of test
tube, glass slides,
cover slip
Red flame – red
hot…wire loops,
straight wires, tip of
forceps, spatula

1. OPEN FLAME
Burning the organism into ashes
incineration

For
garbage
disposal
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Oxidizes cell components &
denatures proteins

HOT AIR OVEN

 Dry heat ovens

Less efficient than moist heat, requiring

longer times & higher temp.

Other name: hot air

oven
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Cell damage depends on the wavelength of the radiation

RADIATION
 RADIATION

Types:
1.UVL or
2.Ionizing
ultraviolet
radiation
light
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Most effective wavelength is 240 – 280 nm.

UVL (ULTRAVIOLET LIGHT)

UVL (ULTRAVIOLET LIGHT)

Poor penetrating ability

Limited to sterilization of
small enclosed area like
operating room & hosp wards
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Greater penetrating ability than UVL

IONIZING RADIATION
chemically interact w/ proteins & nucleic
acids resulting in cell death

Hazardous
to human
cell

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>Deprive m.o. of moisture.
>For food preservation

DESSICATION
 DESSICATION

For preparation of dried fish & nuts

For food
preservation
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Lyophillization (freeze-drying) organism is rapidly frozen, then
dehydrated in high vacuum & stored in a vacuum sealed container

FREEZING
 Freezing

Not reliable. Most m.o. Is resistant to

low temperature

Ex. Large laboratories for

preserving microorganism

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 Filtration

Form of mechanical sieving; m.o. are

not killed but just separated from fld.

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 Filtration

m.o. Not killed, just separated

Filter with 0.22 um pore size

...except mycoplasma,
rickettsia & chlamydia
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 OSMOTIC PRESSURE
Based on principle of
osmosis.
Ex. concentration of
fld surrounding
preservation of m.o. Is
foods in syrup & altered...bacterial
meats in brine cell colapse.

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Denatures proteins. Relatively fast, reliable, safe & inexpensive

MOIST HEAT
 1.Moist heat

Pressurized
boiling pasteurization stream
(autoclaving)

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 boiling
•Temp of 80 –
100 C
Boiling •destroys
for 5 most m.o. &
viruses
mins •except
endospores
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boiling

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 pasteurization

Decreases heat-sensitive m.o. including spoilage

microbes & pathogens (except sporeformers)

UHT (ultra-high
temperature): milk
Heat at 60-65C is exposed to very For milk, milk
followed by rapid high temp for a products &
cooling few seconds. beverages
Stored w/o
refrigeration

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 pasteurization

UHTat(ultra-high
Heat temperature):
60-65C followed by rapid
cooling
milk is exposed to very high
temp for a few seconds.
Stored w/o refrigeration

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 pasteurization

For milk, milk Milk

products & 72 C for 15 mins
beverages

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 Steam under pressure
(autoclaving)

>Destroy all
Other name: forms of
pressurize Most efficient microbes
stream >destroys
endospores

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 autoclaving

15 pound per
Duration: 15 square (psi)
mins to 20 mins; Temperature :
121 C
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 autoclaving
Surgical bandage, media, can
withstand high pressure & temperature

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FRACTIONAL STERILIZATION
(tyndallization)

2 forms of bacteria:
a) heat-labile form-killed by
boiling
b) Heat stable forms –not
killed by simple boiling
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 FRACTIONAL STERILIZATION
(tyndallization)
-boiling ff. by incubating & then
reboiling…repeated several times
• Exposed to live steam for 30 mins for
3 consecutive days

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FRACTIONAL STERILIZATION
(tyndallization)

- Spores –heat resistant

- Germination

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CHEMICAL METHOD OF
STERILIZATION
 FACTORS THAT AFFECT
EFFICACY
Time

NATURE OF ORGANISM (endospore)

Concentration of chem agent (alcohol 50% - 80 %)

TEMPERATURE (high or low)

NATURE OF SURROUNDING MEDIUM (pus, blood)

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TYPES BASED ON
MECHANISM OF ACTION
DAMAGE TO CELL MEMBRANE
interferes active transport & energy
metabolism w/in the cell.

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 1. Surface active agents

Decreases surface tension of cell

membrane lipids
1. Cationic agents Floor, furniture,
• Ex. Quaternary wall
ammonium compound
(zephiran)
• effective at alkaline pH

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 Surface active agents
2. Anionic agents
Removes dirt through emulsification

Most Ex. Soap & Bed linen ,

effective detergents surgical
gown
in acidic
pH
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 2. Phenolic compounds

Distrupt cell wall & precipitate protein

1. Toxic to
phenol humans

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 Phenolic compounds

2. cresol
More For hosp.
potent Envt. /
Safer than hosp.
phenol surfaces
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 3. Alcohol

Disorganize the lipid structure & cause

denaturation of cell proteins

For thermometers,
stet(outer surface), other
equip.
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 1. Ethyl alcohol
Remove
lipid frm
surface; bactericidal
(hand
sanitizer)

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 2. Isopropyl alcohol
Hand
Less volatile; Exerts greater sanitizer,
toxic effects
Has greater (narcosis due disinfection
bactericidal to inhalation
activity of fumes)

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 3. Benzyl alcohol

Used as
preservative

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MODIFICATION OF
FUNCTIONAL GRPS OF
PROTEINS & NUCLEIC ACIDS
Heavy Alkylating
halogens
metals agents

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Heavy metals –damage enzyme
activity of bacteria
• EX. Merthiolate or
1. mercurochrome
MERCURIAL • Biocidal & antiseptic

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 Heavy metals
• Ex. 1% silver nitrate – prophylaxis for
ophthalmia neonatorum
2. Silver • Ex. Silver sulfadiazine – tx for burn
wounds
compd

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HALOGENS – bactericidal oxidizing
agents.Cause inactivation of
enzyme
• Sporicidal, bactericidal, fungicidal,
amoebicidal, viricidal

1. IODINE
• antiseptic

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 halogens
• Common household
2. disinfectant
• For tx. Of water
Chlorine
• For floor, countertops,
blood spills, water tx.

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 Halogens
• For cleaning wound
3. Hydrogen • Weak antiseptic
peroxide

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 ALKYLATING AGENTS
• Ex. formalin
• Spocidal
1. • To kill m.o. Tuberculosis in sputum &
fungi (athlete’s foot)
formaldehyde

• For preserving anatomic specimen,

vaccine prep.
• w/ irritating fumes

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 ALKYLATING AGENTS
• Used as cold sterilant in sterilizing
medical equipment
2. • sporicidal
glutaraldehyde

• For med. equipment

• more potent than formaldehyde

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 ALKYLATING AGENTS
• Used in gaseous
sterilization of heat
sensitive material
3. equipment
• Sporicidal
Ethylene • For tubes
oxide

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 REFERENCES
• Bartolome and Quiles. (2020) Microbiology and
Parasitology: A Textbook and Laboratory Manual for the
Health Sciences. 2nd Edition. C&E Publishing House.
Quezon City. QR65. B37 2020

• • Engelkirk, Paul G. (2019). Burton's microbiology for the

health sciences, 8th ed. . Philadelphia : LWW.616.01 E3
2007

• • Tortora, G.J. (2014) Microbiology. Pearson: Singapore.

616.9041 T63

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