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DEFINITION OF TERMS
Sterilization (Latin sterilis = barren) – killing or removal of ALL viable organisms in an
object or habitat
Disinfection – killing, inhibition or removal of microorganisms that may cause disease
disinfectants: agents used to carry out disinfection and are normally used
only on inanimate object (does not sterilize an object)
Sanitization – reduction of microbial population to levels considered safe by public health
standards
Antisepsis – prevention of infection or sepsis in living tissues using chemicals
antiseptics – generally not as toxic as disinfectants; can be used on living tissues

MICROBIAL CONTROL
Microbial control can be achieved through:
1. inhibition of microbial growth
2. destruction of the microorganism i.e. sterilization
Reasons for controlling microbial growth:
1. to prevent/ limit spoilage or destruction of valuable substances/ commodities
2. to prevent infections
3. to prevent contamination of the cultures, the person and the environment
Agents Used In Microbial Control
Physical Agents
A. Heat
1. moist heat (MOA: denaturation/coagulation of proteins)
a. boiling or flowing steam – kills vegetative cells and eukaryotic spores within
10 minutes
b. pasteurization – process that uses relatively brief exposures to moderately
high temperature to reduce microbial population and to eliminate
human pathogens
1. low temperature holding (LTH) – 62.8°C for 30 mins
2. high temperature short-time (HTST) – 72°C for 15 secs
3. ultra-high temperature (UHT) – 140-150°C for 1 -2 secs
c. steam under pressure (autoclaving) – 121°C for 15 minutes
basis: spores of Bacillus stearothermophilus
d. tyndallization/ fractional steam sterilization/ intermittent sterilization
- for materials destroyed at temperature > 100°C
- sterilization at 90-100°C for 30 mins for 3 consecutive days and
incubated at 37°C in between

MCB 11 – JDPOng
GATBarredo Lecture class – Section D
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2. dry heat
a. direct flame (incineration) – MOA: burning to ashes
b. hot air (mechanical convection oven) – 170 – 180°C for 1 hour
MOA: oxidation of cellular components
B. Low Temperature (MOA: limits growth due to decreased rates of cell reactions and
changes to some proteins)
1. Refrigeration – 4°C
2. Freezing/ Deep freezing – -0° to -95°C
C. Filtration (MOA: exclusion of microorganisms)
used for:
Heat sensitive materials – e.g. enzymes, toxins, vitamins
- Membrane filters
Air – High efficiency particulate air (HEPA) filter removes 99.97% of 0.3 μm
particles
D. Desiccation (MOA: microbiostasis)
1. drying (sun, air, oven)
2. freeze-drying/ lyophilization
E. Increased osmotic pressure (MOA: microbiostasis) – increase solute concentration
eg., by adding sugar, salt
F. Radiation
1. ionizing radiation (MOA: free radicals formation)
- X-rays, gamma rays
- very short wavelength → cause atoms to lose electrons or
ionize
- result in DNA destruction
- excellent sterilizing agent (penetrates deep into objects)

2. non-ionizing radiation (MOA: formation of thymine dimers)

- UV rays
- most lethal at 260 nm → absorbed readily by DNA
- DNA damage
- does not penetrate objects – surface sterilization only

Chemical Agents
Antimicrobial agents: chemicals that kill microorganisms or prevent their growth
-cide (Latin cida = kill) – substances that kill organisms
e.g. Germicide
Bactericide
Fungicide
-static (Greek statikos = causing to stand or stopping) – do not kill but prevents growth
eg. Bacteriostatic/ bacteriostat
Fungistatic/ fungistat

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GATBarredo Lecture class – Section D
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Examples of Antimicrobial Agents

1. Phenol and phenolics (MOA: disruption of plasma membrane, protein denaturation, and
inactivation of enzymes
phenol – used as standard for the effectiveness of other disinfectants
(phenol coefficient)
phenolics – derivatives of phenol
eg. cresols, xylenols, and orthophenylphenol
2. Alcohols (MOA: denature proteins and dissolve membrane lipids)
bactericidal and fungicidal but not sporicidal
eg. ethanol and isopropanol
3. Halogens (MOA: oxidation of cell constituents)
eg. Iodine – can also inhibit protein function
Chlorine
- chlorine gas
- sodium hypochlorite
- calcium hypochlorite
4. Heavy Metals (MOA: denatures enzymes and other essential proteins)
1% silver nitrate → prevents ophthalmic gonorrhea
silver sulfadiazine → used on burns
copper sulfate → algicide
merthiolate → disinfects skin mucous membrane
5. Surface-active Agents (surfactants)
a. soaps and acid-anionic detergents
(MOA: mechanical removal of microorganisms)
eg. bath soaps
b. cationic detergents/quaternary ammonium compounds
(MOA: disrupt plasma membrane and denature
proteins)
6. Organic Acids (MOA: inhibits metabolism)
MOA not related to acidity/pH
widely used in foods/ cosmetics
eg. sorbic acid, benzoic acid, parabens, calcium propionate
7. Sterilizing Gases (MOA:protein denaturation)
used on heat-sensitive items like catheters, plastic Petri dishes and syringes
rapidly penetrate packing materials
microbicidal and sporicidal
eg. ethylene oxide
8. Aldehydes (MOA: protein inactivation)
chemical sterilants
eg. formalin (37% aqueous solution of formaldehyde) used in vaccines
glutaraldehyde used in medical equipment
9. Oxidizing Agents (MOA: oxidation of cell components)
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eg. hydrogen peroxide (H2O2)

Conditions influencing the effectiveness of antimicrobial agent activity

1. population size – larger population, longer time for the microorganisms to die
2. population composition – microorganisms differ markedly in their susceptibility
eg. bacterial endospore more resistant than vegetative cells
younger cells more readily destroyed than mature organisms
3. concentration or intensity of antimicrobial agent
- often but not always, ↑concentration = more rapid killing of microorganisms
- sometimes, ↓concentration is more effective
eg. 70% ethanol is more effective than 95% ethanol
4. duration of exposure – longer exposure → more organisms are killed
5. temperature – increase in T° at which a chemical acts often enhances its activity
6. local environment – environmental factors surrounding the microorganisms may
either protect or aid in their destruction
eg. microorganisms in acidic foods are readily killed by pasteurization than
microorganisms in neutral foods like milk

Points to remember
1. Few chemical agents achieve sterility; most merely reduce microbial populations to
safe levels or remove vegetative forms of pathogens from objects
2. Rough spectrum of susceptibility of microorganisms to disinfectants:
most susceptible: vegetative bacteria
fungi
lipid-containing viruses
less susceptible: mycobacteria
non-lipid containing
viruses
generally resistant: spores

MCB 11 – JDPOng
GATBarredo Lecture class – Section D