CONTENTS

CHAPTER # TOPIC NAME PAGE NO

Chapter – 1 Introduction To Microbiology 2---3

Chapter – 2 Microscope 4---8

Chapter – 3 Bacteria Compared With Other Microorganism 9---11

Chapter – 4 Structure Of Bacterial Cells 12---15

Chapter – 5 Microbial Nutrition 15

Chapter – 6 Growth Cycle 16---17

Chapter – 7 Classification Of Medically Important Bacteria 18---21

Chapter – 8 Normal Flora 22---23

Antimicrobial Drugs, Classification And Modes Of
Chapter – 9 24---28
Action
 Chapter – 1
Introduction to Microbiology
A Brief History of Microbiology

Disease has always left people surprised and terrified. The link between
microorganisms and disease was discovered by Pasteur and Koch. They followed an
explosion of research and discovery that lead to control of many diseases. A brief
history of microbiology will take you through the journey of microbiology from past to
future of microbiology.
Microbiology has had a long, rich history, initially centered in the cause on infectious
diseases but now including practical applications of the sciences. Many individuals have
made significant contribution to the development of microbiology.

Early History of Microbiology: Historians are unsure who made the first observation
of microorganism, but the microscope was available during the mid-1600s, and an
English scientist named Robert Hooke made key observation. He observed strands of
fungi. In the 1670s Dutch merchant named Antony van Leewnhoeke name careful
observation of microscopic organism which he called microscopic world to scientist and
is regarded as one of the first to provide accurate disruptions of protozoa, fungi, and
bacteria.

Louis Pasteur and the Germ Theory: He performed numerous experiments to

discover why wine and dairy product became sour, and he found that bacteria were to
blame. Pasteur postulated the germs theory of disease, which states that microorganism
are the cause of infectious diseases.
Pasteur’s attempts to prove the germ theory were unsuccessful. However, the Germ
scientist Robert Koch provided the proof by cultivating anthrax bacteria apart from any
other types of organism. He then injected pure cultures of the bacilli into mice and
showed that the bacilli caused anthrax. The procedures used by Koch came to be known
as Koch’s postulates. They are set of principles whereby other organism could be
related to diseases.

Koch’s Postulates:
 The organism must be found in all animals suffering from the disease, but not in
healthy animals
 The organism must be isolated from diseased animal and grown in pure culture.
 The cultured organism should cause diseases when introduced into a healthy
animal.
 The organism must be re isolated from the experimentally infected animal.

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The Development of Microbiology: In the late 1800s and for the first decades of
1900s scientist further develops the germ theory of diseases as enunciated by Pasteur
and proved by Koch. There emerged a golden age of microbiology during which many
different infectious diseases were identified. Many of the etiologic agents to microbial
diseases were discovered during and period, leading to the ability to stop epidemics by
interrupting the spread of perfected. In the 1940s, the electron microscope was
developed and perfected. In the decade, cultivation methods for viruses rapidly. With
the development of vaccine in the 1950s and 1960s, such viral diseases as polio,
measles, mumps, and rubella came under control.

Modern Microbiology: Modern microbiology which is into many fields of human

including development of pharmaceutical products, the quality control methods in food
and dairy product production, the control of diseases causing microorganism in
consumable water, and the industrial applications of microorganism. Microorganisms
are used to produce vitamins, amino acids, enzymes, and growth supplements. They
manufacture many foods, including fermented dairy products (soar, cream, yogurt and
butter milk), as well as other fermented foods such as pickles, breads, and alcoholic
beverages one of the major areas of applied microbiology is biotechnology, the this
discipline, microorganisms are as living factories. They produce pharmaceuticals that
otherwise could not be manufactured. These substances include the human hormone
insulin, the antiviral substance interferon, numerous blood clotting factors and clot
dissolving enzymes and a number of vaccines. Bacteria can be reengineered to increases
plant resistance in insects. Biotechnology when present a major application of
microorganism in the next century.

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 Chapter – 2

MICROSCOPE

The microscope is the most commonly used and very important instruments in the
laboratory. It was invented by Von Leuwenhock in 18th century. It is an instrument by
which we can see small objects and their fine details. It is composed of a system of
different lenses (optical system) which enlarge the image many times. It combines the
principle of an optical system and illumination system to achieve illumination in the
bright field. Bacterial cells are not easy to see with bright field microscopy because:

1) Cells are very small

2) Bacterial are colorless

These are problems are solved by good resolution power and staining procedures used
in laboratory.

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PARTS OF MICROSCOPE:
Microscope is composed of three parts
a. Foot piece
b. Body
c. Eye piece

MICROSCOPE (CONTD)

4. Wave Length: Light travels in waves. The length of a wave of light is the wave
length. As light is composed of seven colors every color has its own wave length.

5. Numerical aperture: It is the ratio between the diameter of the lens its focal length.

6. Resolution: The ability of microscope to distinguish two neighbor points as

separate point is called resolution. The greater the resolution power of a microscope,
the cleared the image would be

If the capsule of bacterium can be seen separate for its cell wall, the resolution of
microscope is good.

LIGHT PATHWAY IN MICROSCOPE

Rays if light from the source are directed to the iris diaphragm. Depending upon the
aperture of the diaphragm appropriate amount of the light passes through the iris to
the condenser lens. The condenser lens the rays (through the hole in the stage)
towards the objet i.e. slide. The light then passes through the objective. The lens of the
objectives enlarges the image. The light ray then passes through the inner tube of the
microscope and is directed towards the lens of eye piece. In case of a binocular
microscope the light form the inner tube is directed towards a prism. This prism
diverges the light into two horizontal pathways. Each of these horizontal pathways is
again converted into vertical direction by another set of prism located just below the
eye pieces.

The lens of the eye piece further magnifies the image. Then the rays of light pass
through the eye piece into the eye of the observer. The lens of the human eye focuses
the image on the retina. This is how the object on the slide is visualized after
magnification.

1. INSTRUCTIONS FOR THE USE OF A MICROSCOPE

a. The bench on which the microscopes is placed should be free of any vibration
b. Adjust the height of the chair according to your convenience
c. If using a binocular microscopes, adjust the two eye pieces according to your
interpupillary distance to get a single image

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 d. Place both your forearms on the table
e. Ensure that objectives are clear and free of any oil or dust
f. Check that eye piece is free of any dust
g. Position the sub stage condenser 1-2 mm below the slide
a. Switch on the light source. In case of a built in lamp give 1-2 minutes at very low
illumination for the filament of the bulb to warm before bringing on full
illumination. The filament of cold bulb will burn if given full illumination at
once.
b. Now place the slide on the stage, swing in place the lower power objective.
Focus with coarse adjustment and then with fine adjustment. Change the
objective to high power and adjust again with fine focus.
c. If oil immersion is required focus the field with low power. Apply a small drop
of cedar wood oil on the slide. Change the objective to oil immersion and adjust
with fine focus. While looking under the oil immersion ensure that the
condenser is fully up and iris diaphragm completely open.

1. COMMON PROBLEMS WITH MICROSCOPY

For the beginners there may be problems in adjusting or focusing the microscope.
Following points may be noted:

a. Inability to obtain an image with oil immersion:

1. Check the lens and clean it with lens paper soaked in specific lens cleaner and
the with a lint free cloth.
2. Ensure that the oil is not too thick
3. Check that slide is placed upright
4. Clean the slide with xylol and start again
5. If all the above mentioned steps fail, replace the oil immersion objective with
another. If a clear image is obtained send the objective to manufacture for
replacement.

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 MICROSCOPE (CONTD)
Foot piece
This is the base of the microscope. It gives stability to the body. In light microscopes the
foot piece carries a light source. This consists of a transformer, bulb, a fuse and
sometimes a convex lens to concentrate light. It also has the on/off button for the lamp.

Body
It is part between the foot piece and the eye piece.
a. Nose piece
b. Stage
c. Sub stage

a. Objectives: The body has nose piece which consist of different objectives of
different power. Each objective is engraved with some numerical information and is
also coloured differently for example one objective may read:
Plan 40/0.65
160/0.17
This indicates that it has a x 40 magnification at a tube length of 160 mm and has a
numerical aperture of 0.65. A cover glass of 0.17mm thickness should be used. The
lowest power objective, called scanner, has a red line on it, dry low power (x10)
objective has a yellow line, dry high power (x40) objective has a blue line and oil
immersion objective (x100) has white line on it. These are fixed to a rotary dies.

b. Stage: The stage of a microscope is meant for keeping the slide to be examined. It is
provided with mechanism which not only holds the slide in place but can move it in
two planes by a pair of screws. In most of the microscope the stage has ruled
markings on both axes. These serves as grid reference. For examples while scanning
a slide for acid fast bacilli (AFB) note the reading o the ruled area on both axes if
these are found at a point. With the help of this reading same spot can be located
easily afterwards.

c. Sub-stage: Below the stage is the sub-stage which contains the condenser. This
focuses the light from the light source on the object 9slid). The condenser can be
adjusted up and down. In case of unstained slide e.g. while examining a urine
deposit, feces for ova/cysts or motility of an organism the condenser is adjusted at a
lower position to provide less light and more contrast. In stained slides, e.g.
peripherals blood films; it is adjusted at a higher position, 1-2mm below the stage.
The condenser can be titled form its horizontal position by centering screws. It is
important for laboratory worker to know the centering of the condenser with the
help of the screw.

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 MICROSCOPY

There are different modifications of microscopy.

DARK-FIELD MICROSCOPY
In dark-field microscopy the background remains dark, and only the object is
illuminated. A special condenser mounted under the stage of the dark-field microscope
scatters the light and cause it hit the objective from different angles. Some light bounces
off the object into lens to make the object visible, but the surrounding areas appear dark
because it lacks back-ground light.

Use: Dark-field microscopy helps in the diagnosis of disease cause by spiral bacteria
(e.g. Treponema Palladium)

PHASE CONTRAST MICROSCOPY

Special condense and filter split a light beam and throws the light rays slightly out of
phase. The separated beams of light then pass through and around microscopic object,
the used to illuminate details of living cells. Internal details may also be observed which
cannot be seen with light microscope.

Use: Microbiologists can see organisms alive and unstained with the help of phase-
contrast microscope.

FLUORESCENT MICROSCOPY
Microorganism are coated with a fluorescent dye (such as fluorescin) the illuminated
with ultraviolet light energy. The coated micro-organism appear to fluoresce

ELECTRON MICROSCOPY
A microscopy in which a beam if electrons substitutes for the light energy used in other
microscope are called electron microscopy an electron is a type of microscope that uses
electron to illuminated a specimen and create an enlarged image electron microscopes
has much greater resolving power than high microscopes and cane obtain much higher
magnifications

Types:
Two types of electron microscopes are currently in wide use
A) Transmission Electron Microscope (TEM):
One in which an electron beam passes through an ultra-thin slice of an object. It
is used to photography detailed structures within cells.

B) Scanning Electron Microscope (SEM):

One in which an electron bean sweeps across the surface of an object in the
natural state and without sectioning

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 Chapter – 3

Bacteria Compared with other Microorganism

ORGANISMS INVOLVED IN HUMAN DISEASE:

The organism involved in human infectious diseases belongs to five major groups of
organisms: bacteria fungi, protozoa, helminthes and viruses.
The bacteria belong to the prokaryote kingdom, the fungi (years and molds) and
protozoa are members of the kingdom of protists, and the helminthes (worms) are
classified in the animal kingdom (Table 1-1). The helminthes are complex multicellular
organisms that are classified as metazoa within the animal kingdom. Viruses are quite
cells helminthes and the protozoa are commonly called parasites.

Table 1-1, Biological Relationship of Pathogenic Microorganisms.

Kingdom Pathogenic Microorganisms Types of Cells
Animal Helminthes Eukaryotic
Plant None Eukaryotic
Protist Protozoa Eukaryotic
Fungi Eukaryotic
Prokaryote Bacteria Prokaryotic
Viruses None cellular

IMPORTANT FEATURES
One salient feature is that bacteria, fungi, protozoa and helminthes are cellular whereas
viruses are not. This distinction is based primarily on three criteria.
1. Structure. Cells have a nucleus or nucleus or nucleoid, which contains DNA; this
is surrounded by cytoplasm, within which proteins are synthesized and energy is
generated. Viruses have an inner core of genetic material (either DNA or RNA)
but not cytoplasm, and so they depend on host cells to provide the machinery for
protein synthesis and energy generation.
2. Method of Replications. Cell replicate either by binary fissions or by mitosis,
during which one parent cell divides to make two progeny cells while retaining
its cellular structure. Prokaryotic cells, e.g. bacteria, replicate by binary fission,
whereas eukaryotic cells replicate by mitosis. In contrast, viruses disassemble,
produce many copies of their nucleic acid and protein, and then reassemble into
multiple progeny viruses. Furthermore viruses must replicate within host cells
because, as mentioned above, they lack protein synthesizing and energy

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 generating systems. With the exception of rickettsia and chlamydia, which also
require living host cells for growth bacteria can replicate extra cellularly.
3. Nature of the nucleic acid. Cells contain both, Deoxyribonucleic acid (DNA) or
Ribonucleic acid (RNA), whereas viruses contain either DNA or RNA but not
both.

1. For comparisons, a human red blood cell has a diameter of 7µm

2. Yeasts divided by budding, whereas molds divide by mitosis
3. Helminthes cells divide by mitosis, but the organism reproduces itself
complex sexual life cycles.
*
EUKARYOTES & PROKARYOTES
Cells have evolved into two fundamentally different types, eukaryotic and
prokaryotic, which can be distinguished on the basis of their structure and the
complexity of their organization.

Fungi and protozoa are eukaryotic, whereas bacteria are prokaryotic.

Table 1-2 Comparison of Medically Important Organism

Protozoa and
Characteristic Viruses Bacteria Fungi
Helminthes

Cells No Yes Yes Yes

Approximate 15-25
0.02-0.2 1-5 3-10 (yeasts)
Diameter (µm)1 (trophozoites)
Either DNA or Both DNA and Both DNA and Both DNA and
Nucleic Acid
RNA RNA RNA RNA
Type of Nucleus None Prokaryotic Eukaryotic Eukaryotic

Ribosomes Absent 70S* 80S* 80S

Mitochondria Absent Absent Present Present

Protein
Rigid wall Rigid wall
Nature of outer capsid and Flexible
containing containing
surface lipoprotein membrane
peptidoglycan chitin
envelop
Motility None Some None Most
Method of Not Binary Budding of
Binary Fission Mitosis
Replication Fission mitosis

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 1. The eukaryotic cell has a true nucleus with multiple chromosomes surrounded
by a nuclear membrane and uses a mitotic apparatus to ensure equal allocation
of the chromosomes to progeny cells.
2. The nucleoid of a prokaryotic cell consists of a single circular molecule of loosely
organized DNA, lacking a nuclear membrane and mitotic apparatus (Table1-3).

Table 1-3 Characteristics of Prokaryotic and Eukaryotic Cells

Prokaryotic Eukaryotic Human
Characteristic
Bacterial Cell Cells
DNA with in a nuclear membrane No Yes
Mitotic division No Yes
DNA associated with histones No Yes
Chromosome number One More than one
Membrane-bound organelles, such as
No Yes
mitochondria and Lysosomes
Size of ribosome 70S 80S
Cell wall containing peptidoglycan Yes No

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Motility is another characteristic
By which these organisms can be distinguished

Most protozoa and some bacteria are motile, whereas fungi and viruses are nonmetric

The protozoa are a heterogeneous group that possess three different organs of
locomotion, flagella, cilia and pseudopodia

The motile bacteria move only by means of flagella

Examples of Organism
1) Bacteria : Salmonella typhi
Klebsiella pneumoniae

2) Virus : Hepatitis B virus

Rota virus

3) Fungus : Candida albicans

Aspergillus fumigatus

4) Protozoa : Entamoebahistolytica
Giardia Lambila

5) Helminth : Ascaris lumbricoides

Ancylostoma duodenale

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 Chapter – 4

Structure of Bacterial Cells

Introduction:
 Bacteria are microscopic (very tiny) organisms that are unicellular (made up of a
single cell). Bacterium is the term for single bacteria.
 Evolution of Bacteria: The primitive organism was among the first to appear on
Earth, Bacteria evolved roughly 3.5 billion years ago.
 Discovery of Bacteria: Bacteria were unknown people until the 1600s, when
Antony Van Leeuwenhock first observes them in his newly-made microscope.
 Cells of Bacteria: The cells of bacteria are different from those of plant and animals
in many ways, the most obvious of which is that bacteria lack on nucleus and other
membrane bound organelles (except ribosome’s). Unlike animals and plant, bacteria
have Pilli, flagella and most have a cell capsule.
 Structure of Bacteria Cells: Typical structure of a bacterium is illustrated in figure
and the important features of each component are presented in Table 2-1

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Table 2-1 Bacterial Structures

Structure Chemical Composition Function

Gives rigid support, protects
Essential Components
against osmotic pressure is the
Cell Wall Sugar backbone with peptide
site of action of penicillin’s and
Peptidoglycan side chains that are cross-linked
cephalosporin and is degraded
by lysozyme
Lipid A
Outer membrane of gram-
Major surface antigen used
negative bacteria
frequency in laboratory
Polysaccharide
diagnosis.

Surface fibers of gram- Teichoic Acid

Major surface antigen but rarely
positive bacteria
used in laboratory diagnosis
Lipoprotein bilayer without Site of oxidative and transport
Cytoplasmic Membrane
sterols enzymes
Protein synthesis, site of action
RNA and protein in 50S and 30S of aminoglycosides,
Ribosome
subunits erythromycin, tetracycline’s and
chloramphenicol
Nucleoid DNA Genetic Material
Invagination of plasma Participants in cell division and
Mesosome
membrane secretion
Space between plasma Contains many hydrolytic
Periplasm
membrane and outer membrane enzymes, including β-lactamases
Nonessential
components
Polysaccharide Protects against phagocytosis
Capsule
Two types (1) mediates
attachment to cell surface (2) sex
Pillus or fimbria Glycoprotein
pilus mediates attachment of
two bacteria during conjugation
Flagellum Protein Motility
Provides resistance to
Spore Kerantinlike coat; dipicolinic acid
dehydration, heat and chemicals
Contains a variety of genes for
Plasmid DNA
antibiotic resistance and toxins
Granule Glycogen, lipids, polyphosphates Site of nutrients in cytoplasm

Glycocalyx Polysaccharide Mediates adherence to surface

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Some different factors among Gram-positive and Gram-negative bacteria are given below:

Gram-Positive Bacteria Gram-Negative Bacteria

1) Have 3 layers
1) Have 2 layers i) Inner cytoplasmic membrane
i) Inner Cytoplasmic Membrane ii) Middle thin peptidoglycan layer (5-10%)
ii) Outer Thick peptidoglycan layer %60 iii) Outer Membrane with lipopolysaccharide
(LPS), lipoprotein and phospholipids
2) Low Lipid content 2) High Lipid Content
3) Periplasmic space present between
cytoplasmic membrane and peptidoglycan layer.
3) Periplasmic space absent It contains
β-lactamases (enzymes) that degrades
β-lactamases antimicrobials
4) Endotoxins absent 4) Endotoxins Present

5) Porin channels absent 5) PorinChanneslpresnt

6) Fibers of Lipo-teichoic acid (LTA) present
that protrude outside the peptidoglycan 6) Absent in Gram-Negative Bacteria
layer

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 Chapter – 5

MICROBIAL NUTRITION

In order to identify and study bacterial specie it is necessary to grow the organism
under Laboratory Condition and it is therefore, essential to know its growth
recruitment.

The growth of micro-organisms is dependent on an adequate supply a suitable

nutrients. Some micro-organisms are able to grow under a wide range of condition e.g.
E coil.

Some require extra-nutrition and different environment condition to grow e.g.

gonococcus

Major Nutrient Requirements:

The nutrients required by microorganism are according to their biosynthetic abilities.
The main elements required for growth are carbon, hydrogen oxygen and nitrogen with
sulpher, phosphorous required in somewhat smaller amount and other elements such
as sodium, potassium, magnesium, iron and magnese in small amount. Hydrogen and
O2can be supplied in water. Carbon and nitrogen are the main bulk elements required.

Carbon & Energy Source:

Parasitic micro-organisms normally use organic compounds such as carbohydrates and
amino acid present in the tissue fluids of their host. They are the building blockers of
the organism and provide energy

Nitrogen Source:
Ammonia is the main nitrogen source used in Biosynthesis of micro-organism. It can be
provided directly in the environment or it can be produce indirectly by the organic
nitrogenous nutrients such as amino acids or by the nitrates A few bacteria can use
gaseous nitrogen as a nitrogen source.

Organic Compounds:
Certain organic compounds such as amino acids, nucleotide, monosaccharide, lipid and
co-enzymes, lipid are synthesized by the microorganism of provided by environment
they cannot be synthesized by all micro-organisms. Certain bacteria requires growth
factor or microbial vitamin e.g. thiamine, riboflaxin, nicotine acid, pyridoxine, P-
aminobenzoic acid, folic acid, brotin, cobamideetc

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 Chapter – 6

GROWTH CYCLE

 Bacteria reproduce by binary fission, a process by which one parent cell divides to
form two progeny cells. Because on cell gives rise to two progeny cells. Bacteria are
said to undergo exponential growth(logarithmic growth). The concept of
exponential growth can be illustrated by the following relationship
Number of Cells 1 2 4 8 16
Exponential 20 21 22 23 24
 Thus, I bacterium will procedure 16 bacteria after 4 generations.
 The doubling (generation) time of bacteria ranges form as title as 20 minutes for
Escherichia coli to more than 24 hours for Mycobacterium tuberculosis.
 The exponential growth and the shot doubling time of some organisms result in
rapid production of very large numbers of bacteria. For example, 1E coli.Organism
will produced over 1000 progeny in about 3 hours and over 1 million in about 7
hours.
 The doubling time varies not only with the species but also with the amount of
nutrients, the temperature the pH, and other environment factors.
 The growth cycle of bacteria has four major phases
 If a small number of bacteria are inoculated into a liquid nutrient medium and the
bacteria are counted at frequent intervals, the typical phases of a standard growth
curve can be demonstrated.
 The first is the lag phase, during which vigorous metabolic activity occurs but cells
do not divide. This can last for a few minutes up to many hours.
 The log (logarithm) phase is when rapid cell division occurs.
 The stationary phase occurs when nutrient depletion or toxic products cause growth
to slow until the number of new cells produced balance the number of cells that die
resulting in a steady state.
 The final phase is the death phase, which is marked by a decline in the number of
viable bacteria.

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Aerobic & Anaerobic Growth
For most organisms, an adequate supply of oxygen enhances metabolism and growth.
The oxygen acts as the hydrogen acceptor in the final steps of energy production
catalyzed by the flavoproteins and cytochromes. Because the use of oxygen generates
two toxic molecules, hydrogen peroxide, (H2O2) and the free radical superoxide (O2),
bacteria require two enzymes to utilize oxygen. The first is superoxide dismutase, which
catalyzes the reaction.

2O2+2H2--------H2O2+O2

And the secondo is catalase, which catalyzes the reaction

2H2O2-----------2H2O+O2

The response to oxygen is an important criterion for classifying bacteria and has great
practical significance because specimens from patients must be incubated in the proper
atmosphere for the bacteria to grow.

1. Some bacteria, such as M. tuberculosis are obligate aerobes that is they require
oxygen to grow because their ATP- generating system is dependent on oxygen as
the hydrogen acceptor
2. Other bacteria such as E.coil are facultative anaerobes:they utilize oxygen to
generate energy by respiration if it is present, but they can use fermentation
pathway to synthesize ATP in the absence of sufficient oxygen.
3. The third group of bacteria consists of the obligate anaerobes, such as
Clostridium tetani, which cannot grow in the presence of oxygen because they
lack either superoxide dismutase some, can survive but are not able to grow,
whereas other are killed rapidly.

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 Chapter – 7

CLASSIFICATION OF MEDICALLY IMPORTANT BACTERIA

Classification: It is the process of systematically dividing organism’s into groups, with

the species being the smallest and most definitive level of division.

Simple criteria for the purpose of bacterial Classification include many properties for
e.g.:

 Gram stain reaction (Colour of the dye retained by the bacteria)

 Morphology (shape) of bacteria on microscopy
 Presence of absence of specialized structure like spore, flagella etc
 Hemolysis on sheep blood agar (SBA)
 Production of characteristics pigments e.g. green pigment produce by
pseudomonas aeruginosa
 Colony morphology on different agar media e.g. MacConkey’s agar Lactose
fermenting and non-Lactose fermenting bacteria

Gram stain reactions combined with basic morphology of bacteria under light
microscope are simple and effective methods of classifying many medically important
bacteria. It helps a microbiologist to identify microorganism quickly.
Christian Gram, a histologist in 1884 develops the procedure of staining the bacteria.
Bacteria are classified as Gram positive and Gram negative according to their response
to Gram staining

Gram Positive ________________________________ Appear purple

Gram Negative ______________________________ Appear pink

Classification of Bacteria According to Morphology

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 Chapter – 8

NORMAL FLORA

INTRODUCTION:
It is the term used to describe various Micro-organisms living as permanent resident of
the human body various bacteria and fungi are members of normal flora. (Normal
microbiota). Viruses and Parasites are usually not considered members of normal flora.
Normal flora is usually beneficial. When the micro-organisms are found at other site i.e.
other than usual places they are called pathogen. Norma floras are usually beneficial.
When they change the sire and are found at other places they are termed Pathogenic.
For example, when bacteroides bacteria, which normally reside in the intestine, may
produce abscesses if they penetrate into deeper tissue.E.coli. a normal inhabitant of the
gastrointestinal tract is the most common cause or urinary tract infections. An
opportunistic pathogen causes disease in a host that is physically weak or debilitated.
The members of the normal flora are the permanent resident and the transient
microbiota are the members of the normal flora are not always present or a are present
for only a few days, weeks, or months before disappearing. There is also a disinfection
to be made between members of the normal flora and the colonization of the individual
with a new organism. The term “Colonization” typically refers to the acquisition of a
new organism which colonize (i.e. attaches and grows, usually on a mucosal
membrane), it may cause an infectious disease or it may be eliminated by our host
defenses. The person colonized by a new organism can transmit that organism to other,
i.e. act as a reservoir or infection for others.

SIGNIFICANCE OF THE NORMAL FLORA:

Normal flora contributes to our existence in several ways and plays role both in the
maintenance of health and in the causation of diseases.

1) They may prove beneficial to human e.g. The intestinal bacteria (e.g. bacteroides)
produce vitamins (such as vit B and K). They may break down food stuffs that are
normally ingested by the host into components.
2) The normal flora helps us by competing with pathogens and therefore protecting
the host.

ANATOMIC LOCATIONS OF NORMAL FLORA:

Human body is host for the billions of bacteria of many different kinds. These bacteria
exist in many different parts of the body according to the nature of the local
environment, which is determined by pH, temperature, oxygen, water and nutrient
levels. Other factors such as peristalsis, saliva, lysosome secretion, and immunoglobulin
also play important roles in flora control. Few examples of sites where normal flora is
found are as under.

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 1) Skin
2) Oral cavity
3) Gastrointestinal tract.
4) Upper respiratory tract.

Location Important Organisms Less Important Organisms

Staphylococcus aureus,
Corynebacterium (diphtheroids),
various streptococci, Pseudomonas
Skin Staphylococcus Epidermidis
aeruginosa, anaerobes
(egPropionibacterium,), yeasts (eg.
Candida albicans)
S.epidermidis, Coryncbacterium
Nose Staphylococcus aureus
(diphtheroids), various streptdo
Various streptococci,
Mouth Viridans streptococci
Eikenellacorrodens
Prevotellaintermedia
Dental plaque Streptococcus mutans
Porphyromonasgingivalis
Various anaerobes, eg.
Gingival
Bacteroides, Fusobacterium.
Crevices
Streptococci, Actinomyces
Various streptococci (including
streptococcus pyogenes and
Throat Viridans Streptococci streptococcus pneumonia), Neisseria
specles, Haemophilus influenza,
S.epidemidls
Bifidobacterium, Eubacterium,
Fusobacterium, Lactobacillus, Various
Bacteroides fragilis, Escherichia
Colon aerobic gram-negative rods,
coli
Enterococcus faecalis and other
streptococci, Clostridium
Various, streptococci, various gram-
Lactobacillus, E.coli group B negative rods, B,
Vagina
streptococci fragilisCorynebacterium
(diphtheroids), C.albicans
S.epidermidis, Corynebacterium
Urethra (diphtheroids), various strepto various
gram-negative rods, eg. E.coli

[General Microbiology Reading Material] Page 24

 Chapter – 9

ANTIMICROBIAL DRUGS, CLASSIFICATION AND MODES OF ACTION

INTRODUCTION OF ANTIBIOTICS:

Definition
Substance produced by micro-organism that inhibit the growth or life of other in high
concentration is called “Antibiotics”

The most famous antimicrobial penicillin was discovered by Sir Alexander Fleming in
1929.

Antimicrobial drugs are used for the treatment of infectious diseases because of their
selective toxicity, i.e. the ability to kill an invading micro-organism without damaging
the cells of the host.

BASIC DEFINITIONS:
1) BACTERIOSTATIC DRUGS:
Inhibit: the growth of bacteria & their Multiplication

2) BACTERICIDAL DRUGS:
Kill bacteria

3) NARROW SPECTRUM:
Antimicrobial agents acting only on a single or a limited group of microorganisms.
e.g. Isoniazid is active only against Mycobacteria

4) BROAD SPECTRUM;
Antimicrobials that affect a wide variety of microorganism
e.g. Tetracycline and Chloramphenicol

CLASSIFICATION OF ANTIMICROBIAL AGENTS:

a. Chemical structure (B-Lactams, aminoglycosides)
b. Mechanism of action (Cell Wall Synthesis inhibitors)
c. Activity against particular types of organism (bacteria, fungi, viruses)
d. Site of Action: There are four major sites in the bacterial cell that serve as
a target for antimicrobial agent i.e. cell wall, ribosomes
(protein)

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[General Microbiology Reading Material] Page 26
INHIBITORS OF CELL WALL SYNTHESIS
Some antimicrobial drugs selectively interfere with the synthesis of the bacteria cell
wall. They act by bindings to specific receptors in cell wall. e.g. (β-Lactam antibiotics)
and other examples are Vancomycin and Bacltracin

[General Microbiology Reading Material] Page 27

Β-Lactamase Inhibitor:
Clavulanic acid, sulbactam and tazobactam bind to and inactive β-Lactamases, thereby
protecting the antibiotics that are normally degraded by the enzymes.

Antimicrobials Acting on Ribosomes

They act by inhibiting protein synthesis.

Figure:
Summary of Proteins Synthesis Inhibitors

Antibiotics that inhibit Nucleic acid (DNA, RNA) or Precursors of RNA and DNA

Fluroquinolones, Quninolones and Urinary tract antiseptics (Nitrofurantoin) act on

bacterial DNA (Deoxy Ribonucleic acid) thus inhibiting the growth of bacterial.
Important drugs are classified in the figure below.

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Some groups of antimicrobials inhibit folate which helps to form amino-acids
(precursors of RNA and DNA) that required for bacterial growth. These antimicrobials
are classified below:

Inhibitors of RNA synthesis:

Inhibits RNA Synthesis by inhibiting RNA polymerase (e.g. Rifampicin)

Inhibitors of Cell Membrane Function

Two important antimicrobials polymyxin B and Colistin (Polymyxin E) makes pores in
the cell membrane of bacterial leading to lysis of the organism.

[General Microbiology Reading Material] Page 29