Unit #2

control of micro-organism
By
NAZIA Khan Pitafi
[Link]

Nursing instructor
SUBJECT: MICROBIOLOGY
BSN (GENERIC) 1ST SEMSETER 1ST YEAR
Objectives:
Explain importance of the Control of Microorganisms growth
Describe some physical and Chemical methods Control of
microbial growth
Define the terms sterilization, antiseptic, asepsis, aseptic,
macrobiotics, microbiocidal antibiotics.
Differentiate between broad spectrum and narrow spectrum
antibiotics.
 Simple and Gram staining of Bacteria
Microbial Genetics
Control of microorganisms by Agents
Gram Staining
Anti microbial agents in therapy
Antibiotics sensitivity
Introduction
The control of microbial growth may involve
sterilization, disinfection, antiseptic ,sanitization.

Sterilization is the destruction of all forms of

microbial life, with particular attention to bacterial
spores.
IMPORTANT DEFINITIONS
Sterilization : it is process in which to kill or destroy the all
living microorganisms by physical or chemical methods.
Disinfection :. Disinfection describes a process that
eliminates many or all pathogenic microorganisms, except
bacterial spores, on inanimate objects .
 In health-care settings, objects usually are disinfected by
liquid chemicals or wet pasteurization.
Disinfectant : An agent that kill microorganisms by applying
to in animate(non living objects.
Antiseptic : A chemical used to reduce the numbers of
microbes on body surfaces or prevention of ourselves from
sepsis and we use dettol ,Savlon and sprit etc
an agent that kill microorganisms by applying living things.
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Sanitization: treatment of food handling equipment used to
meet public health standard and minimize chances of
disease [Link] soap & hot water.
Sepsis :Sepsis is a life-threatening condition that arises when
the body's response to infection causes injury to its own
tissues and organs. It is frequently a final common pathway to
death for many infectious diseases worldwide
Asepsis: A state of being free from pathogen micro organism
Aseptic : Free from microorganism or sepsis .sterilization is
aseptic technique to kill micro organism.
macrobiotics,
microbiocidal
Antibiotics: chemical substance produced by a living
organism, generally a microorganism, that is detrimental to
other microorganisms.
Germicide : an agent to kills the germs
Bactericide : an agent to kills the bacteria
Virucide: an agent to inactivate the viruses
Fungicide: an agent to kills the fungi
Sporicide: an agent that kills bacterial endospores
and fungal spores.
Bacteriostatic Agent: an agent that inhibit the
growth of bacteria
Physical methods of microbial control
Heat
Dry heat:
Direct flaming : used to sterilize inoculating loops and
needle.
Incineration: effective way to sterilize disposable items
(paper, cups, dressing) and biological waste.
Dry heat
Filtration: removal of microbes by passage of
a liquid or air through a screen like material
with small pores. Used to sterilized heat
sensitive material like vaccine, enzymes,
antibiotics and some culture media.
Filtration
Physical methods of microbial control
Moist Heat: kills microorganism by coagulation their proteins.
In general heat moist is much more effective than dry heat
Pasteurization
Refrigeratin
Freezing
Lyophilization
Desiccation
Osmotic pressure
Radiation (ionizing radiation andultravoilet light
Boiling
continue
Moist heat.
Kills microorganisms by coagulating their proteins. In
general, moist heat is much more effective than dry
heat.
Boiling:
Heat to 100°C or more. It Kills 30/28:03 vegetative
forms of bacterial pathogens, almost all viruses, and
fungi and their spores within 10 minutes or less.
Endospores and some viruses are not destroyed
quickly. However brief boiling will kill most
pathogens.
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Pasteurization: It is used to reduce microbes
responsible for spoilage of beer, milk, wine, juices, etc. u
Classic Method of Pasteurization: Milk is heated to 62°C
for 30 minutes. u High Temperature Short Time
Pasteurization (HTST): Milk is exposed to 72°C for 15
seconds. u Ultra High Temperature Pasteurization
(UHT): Milk is treated at 140°C for 3 seconds and then
cooled very quickly in a vacuum chamber. U Advantage:
Milk can be stored at room temperature for several
months.
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Refrigeration: Temperatures from 0 to 7°C. It exerts
bacteriostatic effect. Reduces metabolic rate of most
microbes so they cannot reproduce or produce toxins.
Freezing: Items are protected by stopping the growth
of microbes at temperatures below 0°C.
Lyophilization: (Greek, and means "made solvent-
loving"). Lyophilization, commonly known as freeze-
drying, is a way of freezing and then drying something
in a vacuum. Cells and some foods are preserved in
this way.
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.
Radiation
Three 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 and
produce peroxides. 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.
2. Ultraviolet light (Nonionizing Radiation):
Wavelength is longer than 1 nanometer. Damages
DNA by producing thymine dimers. Used to disinfect
operating rooms, nurseries, cafeterias.
 Disadvantages:
Damages skin, eyes. Doesn't penetrate glass, and cloth.
Chemical Methods of Microbial Control
Phenol (carbolic acid): was first used by Lister as a
disinfectant. Destroys plasma membrane and denatures
proteins. 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.
Halogens:
Effective alone or in compounds.
A. lodine: Tincture of iodine denatures proteins.
It Stains skin and clothes.
Continue.

B. Chlorine: Used to disinfect drinking water, pools,

and sewage.
Chlorine is easily inactivated by organic materials.
When mixed in water, forms hypochlorous acid: Cl2 +
H2O H+ + CI+ HOCI Hypochlorous acid
Types of Disinfectants Alcohols:
Kill most bacteria, fungi, but not endospores. Act by
denaturing proteins and disrupting cell membranes.
Evaporate, leaving no residue. Used to mechanically
wipe microbes off skin before injections or blood
drawing. Not good for open wounds, because cause
proteins to coagulate.
Heavy Metals
Heavy metals and their compounds are microbiocidal.
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 sulphate is used to kill algae in pools
and fish tanks.
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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.
Types of Disinfectants 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.
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.
 Importance of Control of Microbial
Growth
Control of microbial growth means to kill or inhibit
the growth of 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.
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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.
 Microbiology
Unit-2
Difference between Broad spectrum and narrow
spectrum antibiotics
Antibiotic: a medicine (such as penicillin or its
derivates) that inhibits the growth of or destroys
microorganisms.
Broad spectrum antibiotics.
Broad spectrum antibiotics is an antibiotic that acts
on the two major bacterial groups, gram positive and
gram negative or any antibiotic that act against a wide
range of disease causing bacteria.
Narrow spectrum antibiotics is an antibiotic that only
kill or inhibit gram positive or gram negative bacteria.
Difference between broad spectrum and
narrow spectrum antibiotics
Broad spectrum antibiotics Narrow spectrum
 Generally affected against  Only effective against a
both gram positive and gram specific type of bacteria
negative bacteria. either gram negative or gram
 Broad spectrum target many positive bacteria.
types of bacteria.  Narrow spectrum target few
 It is used when causative types of bacteria.
agent is not identified.  It is used when causative
 Relatively less safe. agent is identified.
 [Link],  Relatively more safe.
Cefotaxime, Ceftriaxone,  Pencillin ,ampicillin,vancomy
Cephalosporen. cin and vibramycin, Cefaclor.
Gram staining

Gram Staining
Gram stain or Gram staining also called Gram’s method is a method
of staining used to distinguish and classify bacterial species into
two large groups;
Gram-positive bacteria and gram –negative bacteria.
The name comes from the Danish bacteriologist Hans christain
Gram who developed the technique.
Antibiotic sensitivity;
A laboratory test which determines how effective antibiotic therapy
is against a bacterial infections.
Antibiotic sensitivity testing will control the use of antibiotics in
clinical practice.
Testing will assist the clinicians in the choice of drugs for the
treatment of infections.
GRAM POSITIVE VS. GRAM
NEGATIVE

BACTERIA
Most bacteria can be broadly classified as Gram positive
or Gram negative
Gram +ve bacteria have cell walls composed of thick
layers of peptidoglycan.
Gram +ve cells stain purple when subjected to a Gram
stain procedure.
Gram -ve bacteria have cell walls with a thin layer of
peptidoglycan. The cell wall also includes an outer
membrane with lipopolyseccharide (LSP) molecule
attached.
Gram –ve bacteria stain pink when subjected to a gram
stain procedure.
Spectrum of Antibiotics
Narrow spectrum antibiotics: isoniazid (it is known
as isonicotinic acid) is active only against
mycrobacteria.
Extended spectrum: Ampicilline acts against gram
positive and some gram negative bacteria.
Broad Spectrum Antibiotics: tetracycline and
chloramphenicol affect a wide variety of microbial
species.
ANTIMICROBIAL THERAPY
Antimicrobial therapy kills and inhibits the growth of
microorganisms such as bacteria, fungi, protozoan.
Therapies that kill microorganisms are called
microbiocidal therapies.
Therapies that only inhibit that growth of
microorganisms are called microbiostatics
therapies.
Antimicrobial therapy
Empiric
- This antibiotic therapy is often given to patient who have a
proven suspected infection, but where the responsible
Infecting organism have not yet been identified
- More broad spectrum.

Definitive
- Organisms identified and specific therapy chosen.
- More narrow spectrum

Prophylactic or preventive
prevent an initial infection or its recurrence after infection.
Antibiotics prophylaxis is the use of antibiotic usually before
surgery, to prevent a bacterial infection.