Terminology

1. sterilization- is the removal or destruction of all forms of microbial life.

2. Sterilant- a sterilizing agent

3. Commercial sterilization- limited heat treatment used in the food industry

where food is canned.

4. Disinfection- refers to the destruction of vegetative pathogen.

5. Antisepsis- destruction of vegetative pathogen on living tissues with

chemicals called antiseptic.

6. 6. degerming- removal of microbes from a limited area such as the skin

around an injection site.

7. Sanitization- treatment intended to lower microbial counts or eating and

drinking utensils to safe public health levels.

8. Biocide/germicide- kills microorganisms except endospores.

9. Bacteriostasis- treatment that only inhibit the growth and multiplication
of bacteria.

10. Sepsis- the presence of pathogenic organisms in blood and tissue.

11. asepsis- the absence of contamination by unwanted organisms

12. Thermal death point (TDP) is the lowest temperature at which all the
microorganisms in a particular liquid suspension will be killed in 10 mins.

13. Thermal death time (TDT) is the minimal length of time for all bacteria in
a particular liquid culture to be killed at a given temperature.

14. Decimal Reduction time (DRT) is the time in minutes, in which 90% of a
population of bacteria at a given temperature will be killed.
15. Autoclaving- reliable method of sterilization using steam under pressure.

16. Pasteurization- the process of mild heating to kill particular spoilage

microorganisms or pathogens.

17. flaming- method of dry heat sterilization.

18. Hot air sterilization- sterilization by the use of an oven at 170 degrees Celsius
for approximately 2hrs.

19. Incineration- method of heat sterilization where objects burn to ashes.

20. boiling- type of moist heat sterilization that kills vegetative forms of bacterial
pathogens, almost all viruses and fungi and their spores within 10 minutes.

21. Free flowing steam- type of moist heat sterilization that uses the same
temperature as boiling but do not destroy microorganism as fast.
22. filtration- is the passage of a liquid or gas through a screen-
like material with pores small enough to retain
microorganisms.
 The Rate of Microbial Death
- When bacterial population are heated or treated with antimicrobial
chemicals, they usually die at a constant rate.
 Factors influencing the effectiveness of antimicrobial
treatments
1. The number of microbes: the more microbes there are to begin
with, the longer it takes to eliminate the entire population.
2. Environmental Influences: the presence of organic matter often
inhibits the action of chemical antimicrobials.
- The nature of the suspending medium is also a factor in heat
treatment.
3. Time of exposure: chemical antimicrobial often requires extended
exposures for more resistant microbes or endospores to be
affected.
4. Microbial characteristics: gram neg. bacteria are generally more
resistant than gram pos. to disinfectants and antiseptics.
- Mycobacterium, endospores, protozoan cysts, prions are resistant to
disinfection and in the case of prions they are also resistant to
autoclaving.
 Action of Microbial Control Agents
1. Alteration of membrane permeability
- damage to the lipids or proteins of the plasma membrane by
antimicrobial agents causes contents to leak into the
surrounding mediums and interferes with the growth of the
cells.

2. Damage to proteins and nucleic acids

- Breakage of the hydrogen bonds that link adjoining portions
of the amino acid chain results in denaturation of the
protein.
- Damage to the DNA and RNA by heat, radiation, or chemicals
is lethal to the cell making it unable to replicate or carry out
its normal functions such as the synthesis of enzymes.
 Physical Methods of Microbial Control
1. Heat
(a) Moist heat- boiling or steaming
- autoclaving
(b) Pasteurization
(c) Dray heat – direct flaming
- Incineration
- Hot air sterilization
2. Filtration

3. Cold - (a) Refrigeration (b) Deep freezing (c) Lypohilization

4. High pressure
5. Dessication
6. Osmotic pressure
7. radiation
 HEAT
- Laboratory media, glass wares, and hospital instruments, are
usually sterilized by heat.
- Heat appears to kill microorganisms by denaturing their
enzymes.
- Heat resistance varies among different microbes. These
differences can be expressed through the concept of thermal
death point (TDP), thermal death time (TDT) and decimal
reduction time (DRT).
- Moist heat kills microorganisms primarily by denaturation,
which is caused by the breakage of the hydrogen bonds that
hold the proteins in their three dimensional structure. Eg. Of
moist heat sterilization is boiling or free flowing steam
- Boiling and free flowing steam are not very reliable, reliable
sterilization requires temperatures above the boiling point of
water.
 Autoclaving
- Autoclaving is the preferred method of
sterilization, unless the material can be damaged
by heat or moisture. The higher the pressure in
an autoclave the higher the temperature. Eg if
the temperature is 1000C is places under 1 atm of
pressure, the temperature immediately rises to
1210C.

- Autoclaving is used to sterilize culture media,

instruments, dressing, intravenous equipment
etc.
 Pasteurization
- Pasteurization- heat treatment for milk that kills most non pathogens.

- Many relatively heat resistant (thermoduric) bacteria survive

pasteurization, but these are unlikely to cause disease or cause
refrigerated milk to spoil.

- Products other than milk, such as ice cream, yogurt and beer, all have their
own pasteurization time and temperature. This because heat is less
efficient in foods that are more viscous, and fats in food can have a
protective effect on microorganisms.

- Average pasteurization time for milk is 72 degrees Celsius for 15 sec. This
is known as high temperature short time (HTST) pasteurization. The
process also lowers bacterial count.
- Milk can also be sterilized by ultra high temperature (UHT) so that they
can be stored without refrigeration. In the process the milk is actually
heated by super hot steam of 1400C for 3 sec.

NOTE : increase temperature kills microbes faster

 Dry Heat Sterilization
- Dry heat kills by oxidation effects.
Methods:
1. Flaming
2. Hot air sterilization- items are placed in an oven at 170
degrees for 2hrs. The higher heat and longer period is
required because heat in water is more readily transferred to
a cool body than heat in air.
 Filtration
- Used to sterilized heat sensitive materials, such as some
culture media, enzymes, vaccines and antibiotic solutions.
- Is composed of substances such as cellulose esters or plastic
polymers. They are 0.1 mm thick and the pore size is between
0.22 µm- 0.45 µm
- Works by separating bacteria from suspended liquid.
- It removes microbes by passage of a liquid or gas through a
screen like material.
 cold
1. refrigeration- decrease chemical reaction and possible changes in
proteins.
- Use for food, drugs and culture presentations.
- note: psychrotrophs grows slowly at refrigeration temperature and
will alter the appearance and taste of food over time.
- Rapidly attained subfreezing temperature tend to render microbes
dormant but do not necessarily kill them.
- Slow freezing is more harmful to bacteria because it disrupt the
cellular and molecular structure of the bacteria.

2. Deep freezing- decrease chemical reactions and possible changes

in proteins

3. Lyophilization- decrease chemical reactions and possible changes

in proteins.
 High Pressure
- High pressure alter the molecular structure of the proteins
and carbohydrates resulting in rapid inactivation of vegetative
bacterial cells.
- Endospores are relatively resistant to high pressure. They can
be killed with high temperature or alternating pressure cycles
that cause spore germination.
 Desiccation
- In the absence of water micro organisms cannot grow or
reproduce but can remain viable for years.
- When water is available to them they can resume growth and
reproduction.
- Some bacteria such as gonorrhea bacterium can withstand
dryness only for about an hour but the tuberculosis can
remain viable for months.
- Viruses are generally resistant to desiccation.
- The process is used is used in food presevation
 Osmotic Pressure
- The use of high concentrations of salts and sugars to preserve
food is based on the effects of osmotic pressure.
- High concentration of these substances create a hypertonic
environment that causes water to leave the microbial cell.
- Molds and yeast are much more capable than bacteria of
growing in material with low moisture or high osmotic
pressure.
 radiation
• the effect of radiation depends on its wavelength, intensity,
and duration
• Ionizing radiation has high degrees of penetration and exerts
its effects primarily by ionizing water and forming highly
reactive hydroxyl radicals.
- Used for sterilizing pharmaceuticals and medical and dental
supplies
• Ultraviolet radiation, a form of non ionizing radiation, has low
degree of penetration and causes cell damage by making
thymine dimers in DNA that interfere with DNA replication.
- Control of closed environmnet with UV (germicidal) lamp
• Microwaves can kill microbes indirectly as materials get hot.
 Factors related to effective disinfectants
1. Reading the label- indicate what group of organisms the
disinfectant will be effective against.

2. Concentration- concentration affects its action, so it should

be diluted as specified by the manufacturer.

3. The nature of the material being disinfected- note: organic

material or pH interfere with the action of the disinfectant

4. Will the disinfectant easily make contact with the microbe. It

may need to be left on the surface for some time.
 Chemical Agents used to Control Microbial Growth

1. phenols- disrupt plasma membrane and denature enzymes .

- Has a irritating quality and a undesirable odor
2. Phenolic- disrupt plasma membrane and denature enzymes .
- Use on surfaces, instruments, skin surfaces and mucous
membranes
3. Bisphenols- disruption of plasma membrane
- Use as a disinfectant in soap and hand lotion
4. halogens- inhibits protein function and is a strong oxidizing agent
- Chlorine forms hypochlorous acid which is a strong oxidizing agent
that alters cellular components
- Iodine is an effective antiseptic
- Chlorine gas is used to disinfect water and other chloride
compounds are used to disinfect dairy equipment, eating utensils
and household items.
5. Alcohol (ie. Ethanol and isopropanol)- denatures protein
and dissolve lipids. Uses for swabbing the skin before an
injection, used in degerming.
- Effective against endospores or nonenveloped viruses

6. Surface active agents

(a) Soap and detergents- mechanically removal of microbes
through scrubbing
(b) Acid anionic sanitizers- inactivate enzymes
(c) Ammonium compounds- active ingredient is nitrate which
inhibits iron containing enzymes of anaerobes
- Prevent growth of Clostridium botulinum in food
7. Aldehydes (ie. Formaldehyde an dglutaraldehyde) exert their
antimicrobial effect by inactivating proteins.
8. Peroxygens –(ie. Ozone, peroxide and peracetic acid) are
oxidizing agents. They oxidize molecules inside the cell.
- Used on contaminated surfaces, some deep wounds, in which
they are very effective against oxygen sensitive anaerobes..