Control of Microorganisms

Unite 02
By : Hafiz Ahmad Lubab ktk
Institute of Nursing Sciences,
Khyber Medical University , Peshawar
 Lecture Overview

 Introduction to microbial control

 General terminologies regarding microbial control
 Methods of microbial control
Physical Methods
Chemical Methods
Mechanical Methods
 Anti-microbial agents
 Difference between broad spectrum and narrow spectrum antibiotics.

 Physical Method
Physical Methods
 INTRODUCTION

 Control of microbial growth means to inhibit or prevent growth of

microorganisms. Control of microbial growth usually involves the use of
physical or chemical agents which either kill or prevent the growth of
microorganisms.
 Physical agents include such methods of control as high or low
temperature, desiccation, radiation, and filtration.
 Control by chemical agents refers to the use of disinfectants, antiseptics and
antibiotics.

 Physical Method
Physical Methods
 REASONS FOR CONTROLLING
MICROORGANISMS
Reasons for Controlling Microorganisms.
 Prevent contamination.
 Prevent transmission of pathogen
 To prevent decomposition & spoilage of products
 To prevent contamination in aseptic areas, processes like production of
pharmaceuticals by fermentation
 To maintain aseptic condition in operation theaters, filling area of non
sterile pharmaceuticals.
 Physical Method
Physical Methods
 ACTION OF MICROBIAL AGENTS

 Primary targets of microbial control are the microorganisms that can cause
infection or spoilage of food that are constantly present in the external
environment.

 Modes of action fall into two basic categories.

– Alteration of cell walls or cytoplasmic membranes

– Interference with protein and nucleic acid structure

 Physical Method
Physical Methods
 ALTERATION OF CELL WALL AND
MEMBRANES
 Cell wall maintains integrity of cell

– When disrupted, cannot prevent cell from bursting due to osmotic

effects

 Cytoplasmic membrane contains cytoplasm and controls passage of

chemicals into and out of cell

– When damaged, cellular contents leak out

 Viral envelope responsible for attachment of virus to target cell

– Damage to envelope interrupts viral replication.

 GENERAL TERMS

 Sterilization:
Sterilization is the process of destroying all living organisms and viruses. A
sterile object is one free of all life forms, including bacterial endospores, as
well as viruses.

 Disinfection:
Disinfection is the elimination of microorganisms, but not necessarily
endospores, from inanimate (non-living) objects or surfaces.

 Decontamination:
De-contamination is the treatment of an object or inanimate surface to
make it safe to handle.
GENERAL TERMS
 GENERAL TERMS

 Disinfectant:
A disinfectant is an agent used to disinfect inanimate objects but generally
is toxic to use on human tissues.

 Antiseptic:
An antiseptic is an agent that kills or inhibits growth of microbes but is safe
to use on human tissue.

 Sanitizer:
A sanitizer is an agent that reduces microbial numbers to a safe level.
 GENERAL TERMS

 Sepsis:
The presence of Bacteria Contamination.

 Asepsis:
The absence of Bacteria Contamination.

 Aseptic Technique:
Used to prevent microbial contamination of any clinical procedure and
products of pharmaceutical and food industry to keep standards.
GENERAL TERMS
 GENERAL TERMS

 Antibiotic:
Antibiotic is a substance derived from microorganisms or produced
synthetically to kill or inhibit the growth of other microorganisms i.e.
bacteria.

 Chemotherapeutic synthetic drugs:

Synthetic chemicals that can be used therapeutically.
 Cidal:
An agent that is cidal in action; will kill microorganisms .
 Static:
An agent that is static in action; will inhibit the growth of microorganisms.
 GENERAL TERMS

 Bacteriostatic Agent: An agent that inhibits the growth of bacteria e.g.

tetracycline, macrolides, clindamycin etc.

 Bactericidal agent: An agent that kills bacteria e.g. Aminoglycoside,

betalactum, Fluorquinolones etc.

 Germicide: An agent that kills certain microorganism/ germs

 Virucide: An agent that inactivates/ kill viruses.

 Fungicide: An agent that kills fungi.

 Sporicide: An agent that kills bacterial endospores and fungal spores e.g.

Sodium hypochlorite (NaClO).

GENERAL TERMS
METHODS OF MICROBIAL
CONTROL
PHYSICAL METHODS
 PHYSICAL METHODS

Physical methods:

The methods which are used to destroy or remove microorganisms by means

of physical things are known as physical methods.

Examples:

 Temperature

 Radiation

 Refrigeration

 Desiccation
 PHYSICAL METHODS

1: Temperature:

 Microorganisms have a minimum, an optimum, and a maximum

temperature for growth.

 Temperatures below the minimum usually have a static action on

microorganisms. They inhibit microbial growth by slowing down
metabolism but do not necessarily kill the organism.

 Temperatures above the maximum usually have a cidal action, since they
denature microbial enzymes and other proteins. Temperature is a very
common and effective way of controlling microorganisms.
 PHYSICAL METHODS

A. Thermal death point (TDP)

Lowest temperature at which all of the microbes in a liquid suspension will be

killed in ten minutes.

B. Thermal death time (TDT)

The time required to kill all bacteria in a liquid culture at a given temperature.

C. Decimal reduction time (DRT)

The time required to kill 90% of the bacteria in a liquid culture at a given
temperature.
 PHYSICAL METHODS

i. High temperature:

 Vegetative microorganisms can generally be killed at temperatures from

50°C to 70°C with moist heat.

 Bacterial endospores, however, are very resistant to heat and extended

exposure to much higher temperature is necessary for their destruction.

 High temperature may be applied as either moist heat or dry heat.

 PHYSICAL METHODS

B. Moist heat:

Moist heat is generally more effective than dry heat for killing
microorganisms because of its ability to penetrate microbial cells.

Moist heat kills microorganisms by denaturing their proteins (causes

proteins and enzymes to lose their shape).

It also may melt lipids in cytoplasmic membranes.

 Boiling water

 Autoclaving
 PHYSICAL METHODS

Boiling water:

 Boiling is the process of turning a liquid into a vapor and for a certain
atmospheric pressure, the boiling happens at a specific temperature.

 Boiling to 100°C or more. It kill vegetative form of bacterial pathogen,

almost all viruses, and fungi.

 Endospores and some viruses are not destroyed quickly.

 Hepatitis virus: Can survive up to 30 minutes of boiling. However brief

boiling will kill most pathogens.
 PYHYSICAL METHODS

Autoclaving:
 Autoclaving employs steam under pressure.
 Reliable sterilization with moist heat requires temperature above that of
boiling water.
 Example of moist heat is autoclave, a chamber which is filled with hot
steam under pressure .this is preferred method of sterilization for non-heat
sensitive materials in which temperature of steam reaches 121°C at 15 psi
pressure.(pound per square inch).
 Autoclaving is cidal for both vegetative organisms and endospores, and is
the most common method of sterilization for materials not damaged by
heat.
 PYHYSICAL METHODS

AUTOCLAVE:
 PHYSICAL METHODS

b. Dry Heat:

Dry heat kills microorganisms through a process of protein oxidation

rather than protein coagulation. Examples of dry heat include:

 Direct Flaming

 Hot air Sterilization

 Incineration

 Filtration
 PHYSICAL METHODS

 Direct Flaming:

Used to sterilize inoculating loops and needles. Heat metal until it has a red

glow.

 Filtration:

Removal of microbes by passage of a liquid or air through a screen like

material with small pores .used to sterilize heat sensitive material like vaccine,

, antibiotics, and some culture media. Used in operating rooms and burn units

to remove bacteria from air.

 PHYSICAL METHODS

FILTERATION:
 PHYSICAL METHODS

Hot air Sterilization:

Microbiological ovens employ very high dry temperatures: 171°C for 1 hour;

160°C for 2 hours or longer; or 121°C for 16 hours or longer depending on the

volume. They are generally used only for sterilizing glassware, metal

instruments, and other inert materials like oils and powders that are not

damaged by excessive temperature.

 PHYSICAL METHODS

Hot air Sterilization:

 PHYSICAL METHODS

Incineration:

 The process of burning waste material at temperature high to destroy

contaminants.

 Three types of waste to which incineration is applied extensively are

municipal solid waste, hazardous waste, medical waste.

 Incinerators are used to destroy disposable materials by burning.

 Incineration is a method of treating waste which involves the combustion

of the organic substances found in waste materials.

 PHYSICAL METHODS

Incineration:
 PHYSICAL METHODS

c. Pasteurization:

Pasteurization is the mild heating of milk and other materials to kill particular

spoilage organisms or pathogens. It does not, however, kill all organisms. Milk

is usually pasteurized by heating to 71.6°C for at least 15 seconds in the flash

method or 62.9°C for 30 minutes in the holding method.

This is sufficient to kill the vegetative cells of the milk-borne pathogens

(e.g. mycobacterium, salmonella, streptococcus),but not to sterilize the milk.

 PHYSICAL METHODS

 Classic method of Pasteurization:

Milk is heated to 62°C for 30 minutes.

 High Temperature short time pasteurization:

Milk is exposed to 72°C for 15 seconds.

 Ultra high temperature pasteurization:

Milk is treated at 140°C for 3 seconds and then cooled very quickly in

a vacuum chamber.

Advantages: Milk can be stored at room temperature for several months.

 PHYSICAL METHODS

c. Pasteurization:
 PHYSICAL METHODS

ii. Low Temperature:

Low temperature inhibits microbial growth by slowing down microbial

metabolism. Examples include: refrigeration and freezing.

 Refrigeration at 0°C to 7°C slows the growth of microorganisms and

keeps food fresh for a few days. Bacteriostatic effect.

 Freezing items are protected by stopping the growth of microbes at

temperature below 0°C , and keeps food fresh for several months.
 PHYSICAL METHODS
2. Desiccation:
It is the process of removal of water. In the absence of water, microbes cannot
grow or reproduce, but some may remain viable for years. After water
becomes available, they start growing again.
 PHYSICAL METHODS
3. Radiation:
Two types of radiation kill microbes:
1. Ionizing Radiation: Gamma rays, X rays, electron beams, or higher
energy rays. Have short wavelengths (less than 1 nanometer).
 Dislodge electrons from atoms and form ions.
 Cause mutations in DNA .
 Used to sterilize pharmaceuticals and disposable medical supplies. Food
industry is interested in using ionizing radiation.
Disadvantages:
 Penetrates human tissues. May cause genetic mutations in humans.
 PHYSICAL METHODS

2. Ultraviolet light (Nonionizing Radiation):

 Wavelength is longer than 1 nanometer Damages DNA by producing

thymine dimers.

 The greatest antimicrobial activity of UV light occurs at 250 to 260 nm,

which is the wavelength region of maximum absorption by the purine and
pyrimidine bases of DNA.

 DNA replication is inhibited and the organism can not grow.

 Used to disinfect operating rooms, nurseries.

Disadvantages: Damages skin and eyes.

 CHEMICAL CONTROL OF
MICROORGANISMS
Chemical methods of destruction:
 Very few chemical agents actually achieve sterility, but they do reduce the
microbial population to safe levels, destroying pathogen.
 Chemical agents act primarily by one of the three mechanism.
 disruption of cell membrane
 modification of protein
 modification of DNA
 The following table illustrates some of the commonly used chemical
agents.
 CHEMICAL METHODS

1: Phenol (carbolic acid):

 was first used by Lister(1860) as a disinfectant.
 Destroys plasma membrane and denatures proteins.
 Effective against gram-positive staphylococci and streptococci.
 Rarely used today because it is a skin irritant and has strong odor.
Advantages: Stable, persist for long times after applied, and remain active
in the presence of organic compounds.
 Disadvantages :Excessive use in infants may
cause neurological damage.
 CHEMICAL METHODS
2: Halogens:
Effective alone or in compounds.
A. Iodine:
 Tincture of iodine (2% solution of iodine and potassium iodide in ethanol)
denatures proteins.
 It Stains skin and clothes.
 Used as skin antiseptic in surgery
 Not effective against bacterial endospores
B. Chlorine:
 Used to disinfect drinking water, pools, and sewage.
 Chlorine is easily inactivated by organic materials.
 Sodium hypochlorite is active ingredient of bleach
 When mixed in water, forms hypochlorous acid:
 Cl2 + H2O ------> H+ + Cl- + HOCl
 CHEMICAL METHODS

3: Alcohols:

 Kill most bacteria, fungi, but not endospores.

 Act by denaturing proteins and disrupting cell membranes.

 Used to mechanically wipe microbes off skin before injections or blood

drawing.

 Not good for open wounds, because cause proteins to coagulate.

 They are bactericidal, and fungicidal

but not sporicidal.

 CHEMICAL METHODS
4: Heavy Metals:
 Heavy metals and their compounds are microbicidal.
 Include copper, selenium, mercury, silver, and zinc.
 The property of heavy metal to exert biocidal effect is called Oligodynamic
action.
A. Silver:
 1% silver nitrate used to protect infants against gonorrheal eye infections
until recently.
B. Copper:
 Copper sulfate is used to kill algae in pools and fish tanks.
CHEMICAL METHODS
 CHEMICAL METHODS

C. Selenium

 Kills fungi and their spores. Used for fungal infections.

 Also used in dandruff shampoos.

D. Zinc

 Zinc chloride is used in mouthwashes.

 Zinc oxide is used as antifungal agent in paints.

 CHEMICAL METHODS

5: Oxidizing Agents:

A. Ozone:

 It is Used to disinfect water.

 Helps neutralize unpleasant tastes and odors.

 More effective killing agent than chlorine, but less stable and more

expensive.
 CHEMICAL METHODS

B. Hydrogen Peroxide:
 Used as an antiseptic.
 Not good for open wounds because quickly broken down by catalase
enzyme present in human cells.
 Effective in disinfection of inanimate objects.
 Sporicidal at higher temperatures.
 Used by food industry and to disinfect contact lenses.
C. Benzoyl Peroxide:
 Used in acne medications.
CHEMICAL METHODS
 CHEMICAL METHODS

6. Peracetic Acid :

 One of the most effective liquid sporicides available.

 Sterilant : - Kills bacteria and fungi in less than 5 minutes.

 Kills endospores and viruses within 30 minutes.

 Used widely in disinfection of food and medical instruments because it

does not leave toxic residues.
 CHEMICAL METHODS

6. Aldehydes:

 Highly effective molecule that combine with nucleic acids and protein and
inactivate them, probably by crosslinking and alkylating molecules.

 They are sporicidal and can be used as chemical sterilant.

 Formaldehyde ,which is available as a 37% solution in

water(formalin),denature proteins and nucleic acids.

 Glutaraldehyde is used to disinfect hospital ,laboratory equipment,

respiratory therapy equipment.
 MECHANICAL METHODS

1. Filtration:

 Mechanical means of removing microorganisms. The liquid or gas is

passed through a filter with pores small enough to prevent passage of
microbes. This method can be used for substances that are sensitive to heat.

 HEPA filters: removes at least 99.97% of airborne particles 0.3

micrometers (µm) in diameter.

 ULPA filter: remove from the air at least 99.999% of dust, pollen, mold,
bacteria and any airborne particles with a size of 120 nanometers or larger.
HEPA & ULPA FILTER
 MECHANICAL METHODS

2. Osmotic pressure:
 The use of high concentrations of salts and
sugars in foods is used to increase
the osmotic pressure and create a
hypertonic environment.
 The addition of salt to meat creates a
solute-rich environment where osmotic
pressure draws water out of
microorganisms, thereby retarding their
growth.
Note: Staphylococci spp, Yeast are
resistant to high osmotic pressures.
 Importance of control of microbial
growth
 Control of microbial growth means to kill or inhibit the growth

microorganisms. Control of growth usually involves the use of physical or

chemical agents which either kill or prevent the growth of microorganisms.

 The control of microbial growth is necessary in many practical situations,

and significant advances in medicine, agriculture, and food science have

been made through the study of microbiology.

 Importance of control of microbial
growth
Microbial control is very important in the following different aspects.
 Controls infectious diseases.
 Minimizes cross and nosocomial infections
 Prevents surgical complications
 Decreases morbidity and mortality
 Prevents food from spoilage
 Food can be preserved for months
 Improves and ensures the safety of agricultural products
 Ensures the provision of safe drinking water to public
The last objective will be covered in
second part of this unit.