GENERAL AND MEDICAL MICROBIOLOGY

Malshi Omedhya Galappaththi

BSc/ PhD
 Objectives

At the end of the lecture series you will be able to

gain a basic knowledge on,
• What is Microbiology?
• Diversity
• Human immune system
• Defense mechanisms
 What is microbiology

Study of Micro-organisms: Organisms that EXIST as

Single Cells or cell clusters and must be viewed

individually with the aid of a Microscope

What organisms are considered to be
microbial cells and studied in
microbiology
1. BACTERIA
2. FUNGI
3. ALGAE
4. PROTOZOA
5. Viruses(although not a cellular entity but an
intracellular pathogen)
6. Prions (a biochemical anomaly—misfolded proteins)
7. Helminths Worms (multicellular)
What is a Microbe

 Smaller than 0.1mm

 Includes bugs, things, germs, viruses,
protozoan, bacteria, animalcules, small
suckers
Why study Microbiology

 Microbes are related to all life.

 In all environments
 Many beneficial aspects
 Related to life processes (food web, nutrient
cycling)
 Only a minority are pathogenic.
 Most of our problems are caused by microbes
Diversity of Microbes

 Bacteria-single celled prokaryotes

 Protozoa-eukaryotic, single celled, colonial,
many ways of nutrition
 Fungi- absorb nutrients, single celled
filamentous
 Viruses-acellular entities
 Others- worms, insects
Size Comparison of Microbes
 Bacteria

 Prokaryotes
 Peptidoglycan cell
walls
 Binary fission
 For energy, use
organic chemicals,
inorganic chemicals,
or photosynthesis
Figure 1.1a
Bacterial Cell Structure
 Structure of bacterial cells:
Size, Shape, and Arrangement of bacterial cells:

Morphology and arrangement of bacterial cells are criteria

used for classification of bacteria into following groups:

1. Cocci (Singular: coccus).

2. Rods (bacilli), (Singular: rod, bacillus).

3. Vibrios (Singular: vibrio).

4. Spirilla (singular :Spirillum)

5. Spirochetes. (Singular: Spirochaete).

1. Cocci:
These are round or oval bacteria measuring about
0.5-1.0 micrometer in diameter.
When they multiplying, cocci may form pairs,
chains, or irregular groups.

Cocci in pairs are called diplococci, for example,

Meningococci and Gonococci.
Cocci in chains are called streptococci, for
example Streptococcus yogens.
a

Cocci in irregular groups are called staphytococci,

for example, Staphylococcus aureus.
2. Rods (bacilli):

These are stick-like bacteria with rounded, square, or swollen ends. They
measure 1-10 micrometer in length by 0.3-1.0 micrometer in width.
It may arranged in:
A- Chains, for example, Streptobacillus species.
B- Branching chains, for example, Lactobacilli .
C- Mass together, for example, Mycobacterium leprae.
D- Remain attached at various angles resembling Chinese letters, for
example, Corynebacterium diphtheria.
3-Vibrios:
These are small slightly curved rods measuring 3-4 micrometer in
length by 0.5 micrometers in width.
Most vibrios are motile with a single flagellum at one end.
They show a rapid darting motility.
For example: Vibrio cholerae.
4-Spirochetes:
These are flexible, coiled, motile organism, 6-20 micrometer in length.
They progress by rapid body movements.
Spirochetes are divided into three main groups:
A- Treponemes. B- Borreliae. C- Leptospires.
 Microbiological Classification
of Infectious Diseases

 Bacteria are classified by their Gram stain

characteristics.
 Gram staining is the application of a crystal
violet dye to a culture of bacteria. Bacteria that
retain the color of the dye are called Gram
positive; bacteria that don't are Gram negative.
 The Gram stain attaches to peptidoglycan in the
bacterial cell wall.
 In Gram-negative bacteria, the peptidoglycan layer is
protected by an outer membrane.
Microbiological Classification
of Infectious Diseases
Procedure of Gram Staining
 Take a clean, grease free slide.
 Prepare the smear of suspension on the clean slide
with a loopful of sample.
 Air dry and heat fix
 Crystal Violet was poured and kept for about 30
seconds to 1 minutes and rinse with water.
 Flood the gram’s iodine for 1 minute and wash with
water.
 Then ,wash with 95% alcohol or acetone for about 10-
20 seconds and rinse with water.
 Add safranin for about 1 minute and wash with water.
 Air dry, Blot dry and Observe under Microscope.
Interpretation
 Gram Positive: Blue/Purple Color
 Gram Negative: Red/ pink Color
Principle of Gram Staining
 When the bacteria is stained with primary stain Crystal Violet
and fixed by the mordant, some of the bacteria are able to
retain the primary stain and some are decolorized by alcohol.

 The cell walls of gram positive bacteria have a thick layer of

protein-sugar complexes called peptidoglycan and lipid
content is low.

 Decolorizing the cell causes this thick cell wall to dehydrate

and shrink, which closes the pores in the cell wall and
prevents the stain from exiting the cell.

 So the ethanol cannot remove the Crystal Violet-Iodine

complex that is bound to the thick layer of peptidoglycan of
gram positive bacteria and appears blue or purple in color.
 In case of gram negative bacteria, cell wall also
takes up the CV-Iodine complex but due to the
thin layer of peptidoglycan and thick outer layer
which is formed of lipids, CV-Iodine complex gets
washed off.

 When they are exposed to alcohol, decolorizer

dissolves the lipids in the cell walls, which allows
the crystal violet-iodine complex to leach out of
the cells.

 Then when again stained with safranin, they take

the stain and appears red in color.
 Fungi

 Eukaryotes
 Chitin cell walls
 Use organic chemicals
for energy
 Molds and mushrooms
are multicellular
consisting of masses of
mycelia, which are
composed of filaments
called hyphae
 Yeasts are unicellular
 Protozoa

 Eukaryotes
 Absorb or ingest
organic chemicals
 May be motile via
pseudopods, cilia, or
flagella
 Most free, some
parasites
Figure 1.1c
 Algae

 Eukaryotes
 Cellulose cell walls
 Use photosynthesis for
energy (primary
producers)
 Produce molecular
oxygen and organic
compounds
 Metabolically diverse
 Multicellular Animal Parasites

 Eukaryote
 Multicellular
animals
 Parasitic
flatworms and
round worms are
called helminths.
 Microscopic stages
in life cycles.
 Viruses
 Acellular
 Consist of DNA or RNA
core
 Core is surrounded by a
protein coat
 Coat may be enclosed
in a lipid envelope
 Viruses are replicated
only when they are in a
living host cell
 Microbiological Classification
of Infectious Diseases
 Viruses are acellular, obligate intracellular
organisms.
 The complete infectious virus is termed a virion.
 The virion consists of the specific nucleic acid (DNA
or RNA) surrounded by a protein coat (capsid).
 Some viruses are enveloped which means that they
possess a lipoprotein coat that surrounds the capsid and is
acquired from infected host cell membrane.
 Viruses that lack an envelope are “naked.”
Microbiological Classification
of Infectious Diseases

 Viruses are typically classified by:

 Genetic material (DNA vs. RNA)
 Strandedness (single vs. double)
 Size and shape of the capsid and whether its
enveloped or non-enveloped
 Method of replication
Microbiological Classification
of Infectious Diseases
THE HUMAN IMMUNE SYSTEM
What is the immune system?

 The body’s defense against disease causing

organisms, malfunctioning cells, and foreign
particles
 The First Line of Defense
~Skin~
- The dead, outer layer
of skin, known as the
epidermis, forms a
shield against
invaders and
secretes chemicals
that kill potential
invaders
- You shed between 40
– 50 thousand skin
cells every day!
 The First Line of Defense
~Mucus and Cilia~

- As you breathe in,

foreign particles and
bacteria bump into
mucus throughout your
respiratory system and
become stuck
- Hair-like structures
called cilia sweep this
mucus into the throat
for coughing or
swallowing
Don’t swallowed bacteria have a
good chance of infecting you?
 The First Line of Defense
~Saliva~
What’s the first thing you do when you cut
your finger?

- Saliva contains many

chemicals that break down
bacteria
- Thousands of different types
of bacteria can survive these
chemicals
 The First Line of Defense
~Stomach Acid~

- Swallowed bacteria are

broken down by incredibly
strong acids in the stomach
that break down your food
- The stomach must produce
a coating of special mucus
or this acid would eat
through the stomach!
 Escherichia coli
is common and plentiful in all
of our digestive tracts. Why
are we all not sick?
- These bacteria are
technically outside the
body and aid in digesting
material we cannot

- Only if E.Coli are

introduced in an unnatural
manner can they break
through the first line of
defense and harm us
 The Second Line of Defense
~White Blood Cells~

- If invaders actually
get within the body,
then your white
blood cells (WBCs)
begin their attack
- WBCs normally
circulate throughout
the blood, but will
enter the body’s
tissues if invaders
are detected

Video
 https://www.youtube.com/watch?v=cVIEqR1t
9bQ
 White Blood Cells
~Phagocytes~
 These white blood cells are
responsible for eating foreign
particles by engulfing them

 It engulfs foreign bodies by

extending its cytoplasm into
pseudopods (cytoplasmic
extensions like feet),
surrounding the foreign
particle and forming a
vacuole.
The Second Line of Defense
~Interferon~

- Virus-infected body
cells release interferon
when an invasion
occurs
- Interferon – chemical
that interferes with the
ability to viruses to
attack other body cells

What happens to already

infected cells?
White Blood Cells
~T-Cells~
 T-Cells, often called
“natural killer” cells,
recognize infected
human cells and cancer
cells
 T-cells will attack these
infected cells, quickly kill
them, and then continue
to search for more cells
to kill
 The Second Line of Defense
~The Inflammatory Response~

- Injured body cells

release chemicals called
histamines, which begin
inflammatory response
- Capillaries dilate
- Pyrogens released, reach
hypothalamus, and
temperature rises
- Pain receptors activate
- WBCs flock to infected
area like sharks to blood
Two Divisions of the Immune
System
- The efforts of the WBCs known as phagocytes
and T-cells is called the cell-mediated
immune system.
- Protective factor = living cells
- Phagocytes – eat invaders
- T-cells – kill invaders
Two Divisions of the Immune System

 The other half of the immune system is called

antibody-mediated immunity, meaning that
is controlled by antibodies
 This represents the third line of defense in the
immune system
 The Third Line of Defense
~Antibodies~

- Most infections never make it

past the first and second levels
of defense
- Those that do trigger the
production and release of
antibodies
- Proteins that latch onto, damage,
clump, and slow foreign particles
- Each antibody binds only to one
specific binding site, known as an
antigen
 Antibody Production

- WBCs gobble up invading

particles and break them up
- They show the particle
pieces to T-cells, who
identify the pieces and find
specific B-cells to help
- B-cells produce antibodies
that are equipped to find
that specific piece on a new
particle and attach
 Immunity

- New particles take longer

to identify, and a person
remains ill until a new
antibody can be crafted
- Old particles are quickly
recognized, and a person
may never become ill from
that invader again. This
person is now immune.
 https://www.youtube.com/watch?v=lXfEK8G
8CUI

 https://www.youtube.com/watch?v=zQGOcO
UBi6s
 References

• Microbiology by Prescott
• Any Microbiology Book available
• Reliable online sources
• YouTube videos
Summary

• Microbial Diversity
• Human immune system
• Defense mechanisms
Thank you

Questions?
[email protected]