Dr.

Harender Simar
Assistant Professor
Microbiology, MAMC,
Agroha

2/11/2020 1
Introduction:
 Microorganisms – several classes of living beings

 Based on the organization of their cellular

structures, all living cells can be divided into two
groups: Eukaryotic & Prokaryotic

 Eukaryotic cell types - Animals, plants, fungi, protozoans,

and algae

 Prokaryotic cell types - bacteria & blue green algae

Schematic of typical animal (eukaryotic) cell, showing subcellular components.

Organelles: (1) nucleolus (2) nucleus (3) ribosome (4) vesicle

(5) rough endoplasmic reticulum (ER) (6) Golgi apparatus (7) Cytoskeleton
(8) smooth ER (9) mitochondria (10) vacuole
(11) cytoplasm (12) lysosome (13) centrioles
 Characteristics of prokaryotes and eukaryotes
Characteristics Prokaryotes Eukaryotes
Major groups Bacteria, blue Fungi, parasites, other algae,
green algae plants and animals
Nucleus Diffuse Well defined
Nuclear Absent Present
membrane
Nucleolus Absent Present
Ribonucleoprot Absent Present
ein
Cell division Binary fission Mitosis, Meiosis
Chromosome One, circular Many, liner
Extrachromoso Found in plasmid Found in mitochondria
mal DNA 2/11/2020 10:09 AM
 Characteristics of prokaryotes and eukaryotes
Characteristics Prokaryotes Eukaryotes
Cell membrane Doesn’t contain Contain sterols
sterols except in
mycoplasma
Cellular organelles Absent (except Present
like mitochondria, ribosome)
golgi apparatus etc
Ribosome 70s in size 80s in size

Site of respiration Mesosome Mitochondria

Pinocytosis Absent Present

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Size of Bacteria

 Unit of measurement in bacteriology is the

micron (micrometre, µm).
 1 micron = 10-3 = 1/1000 mm
 Bacteria of medical importance
 0.2 – 1.5 µm in diameter
 3 – 5 µm in length
The size does not matter!!
Shape of Bacteria

 Cocci – spherical/ oval shaped major groups

 Bacilli – rod shaped
 Vibrios – comma shaped
 Spirilla – rigid spiral forms
 Spirochetes – flexible spiral forms
 Actinomycetes – branching filamentous
bacteria
 Mycoplasmas – lack cell wall
Arrangement of bacteria: Cocci
Coccus

Tetrad – groups of four

Cocci in pair – Diplococcus

Cocci in chain - Streptococci

Cocci in cluster - Staphylococci Sarcina –groups of eight

Arrangement of bacteria: Bacilli
Other shapes of bacteria

Comma shaped
Spirilla

Spirochetes
Anatomy of a Bacterial Cell
Anatomy of A Bacterial Cell
 Outer layer – two components:
1. Rigid cell wall
2. Cytoplasmic (Cell/ Plasma) membrane – present
beneath cell wall

 Cytoplasm – cytoplasmic inclusions,

ribosomes, mesosomes and nucleus
(DNA-a single circular chromosome)
 Additional structures – plasmid, slime
layer, capsule, flagella, fimbriae (pili),
spores
Structure & Function of
Cell Components
CELL WALL
 Outermost layer, encloses cytoplasm
1. Confers shape and rigidity

2. 10 - 25 nm thick

3. Weigh about 20-25% of

the dry weight of the cell.

4. Composed of complex polysaccharides (peptidoglycan/

mucopeptide) - formed by N acetyl glucosamine (NAG)
& N acetyl muramic acid (NAM) alternating in chains,
held by peptide chains.
Cell Wall
 Cell wall –
4. Carries bacterial antigens – important in virulence &
immunity

5. Chemical nature of the cell wall helps to divide bacteria

into two broad groups – Gram positive & Gram negative

6. Gram +ve bacteria have simpler chemical nature than

Gram –ve bacteria.

7. Several antibiotics may interfere with cell wall synthesis

e.g. Penicillin, Cephalosporins
Gram positive cell wall

The Gram-positive cell wall is composed of a thick, multilayered

peptidoglycan sheath outside of the cytoplasmic membrane. Teichoic
acids are linked to and embedded in the peptidoglycan, and lipoteichoic
acids extend into the cytoplasmic membrane
Gram negative cell wall

The Gram-negative cell wall is composed of an outer membrane linked to thin,

mainly single-layered peptidoglycan by lipoproteins. The peptidoglycan is
located within the periplasmic space that is created between the outer and
inner membranes. The outer membrane includes porins, which allow the
passage of small hydrophilic molecules across the membrane, and
lipopolysaccharide molecules that extend into extracellular space.
Cell Wall
Summary of the differences between
Gram positive & Gram negative bacteria

Property of bacteria Gram Positive Gram Negative

Thickness of wall 20-80 nm 10 nm

Number of layers in wall 1 2
Peptidoglycan content >50% 10-20%
Teichoic acid in wall + -
Lipid & lipoprotein content 0-3% 58%
Protein content 0% 9%
Lipopolysaccharide 0 13%
Sensitive to penicillin Yes Less sensitive
Digested by lysozyme Yes Weakly
• Bacteria With Defective Cell Wall:-
• cell wall synthesis may be inhibited or interfered
by many factors such as
– antibiotics,
– bacteriophages &
– lysozyme.
• Bacteria without or with deficient cell walls are
of four types:-
• Mycoplasma:-
- Naturally occurring bacteria without cell wall.
- It is as an independent bacterial genus.
- They don’t require hypertonic environment for
maintenance & are stable in culture medium.
2) L-forms:-
- Kleinberg Nobel, while studying Streptobacillus
moniliformis, observed abnormal forms of
bacteria & named them L-forms after Lister
institute.
- develops either spontaneously or in the
presence of penicillin or other agents that
interfere with synthesis of cell wall.
- L-forms are more stable than protoplasts &
spheroplasts.
3) Protoplast:-
-Derived from gram positive bacteria.
-contain cytoplasmic membrane & cell wall is totally
lacking.
- produced artificially by lysozyme in a hypertonic
medium,
- these are unstable.

4) Spheroplasts:-
- derived from gram negative bacteria.
- produced in presence of penicillin.
- osmotically fragile & must be maintained in hypertonic
culture medium.
- differ from protoplast in that some cell wall material is
retained.
• Pleomorphism and Involution:-
• Certain species of bacteria exhibit great variation
in shape and size of individual cells called
Pleomorphism.
• Some bacteria show swollen and aberrant forms
in laboratory old cultures and are known as
involution forms.
• Defective cell wall synthesis is often responsible
for development of these two abnormal forms.
Cytoplasmic (Plasma) membrane
 Thin layer 5-10 nm, separates cell wall from
cytoplasm
 Acts as a semipermeable membrane: controls the
inflow and outflow of metabolites
 Composed of lipoproteins with small amount of
carbohydrates
 Ribosomes

• Ribosomes are the sites for protein synthesis.

• Composed of ribosomal RNA (rRNA) and ribosomal
proteins.
• Ribosomes are integrated in linear strands of mRNA to
form polysomes.
• At this site, the genetic codons of the mRNA are
translated into peptide sequences.
• Each 70 S unit consists of a 30 S and a 50S subunits.

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Other Cytoplasmic Components

 Mesosomes –
1. Multi-laminated structures formed as
invaginations of plasma membrane
2. Principal centre of respiratory enzymes
3. Coordinate nuclear & cytoplasmic division during binary fission
4. More prominent in Gram +ve bacteria

 Intracytoplasmic inclusions – reserve of energy &

phosphate for cell metabolism e.g. Metachromatic
granules in diphtheria bacilli
Nucleus

 No nucleolus
 No nuclear membrane
 Genome –
 single, circular double stranded DNA
 Haploid
 Divides by binary fission
Additional Organelles
1. Plasmid –
 Extranuclear genetic elements consisting of DNA

 Transmitted to daughter cells during binary fission

 May be transferred from one bacterium to another

 Not essential for life of the cell

 Confer certain properties e.g. drug resistance, toxicity

Additional Organelles
2. Capsule & Slime layer –
 Viscous layer secreted around the cell
wall.
 Polysaccharide / polypeptide in nature

a) Capsule – sharply defined structure, INDIA INK STAINING

antigenic in nature
• Protects bacteria from lytic enzymes
• Inhibits phagocytosis
• Stained by negative staining using India
Ink
• Can be demonstrated by Quellung reaction
(capsule swelling reaction)

b) Slime layer – loose undemarcated

secretion

Example: K. pneumoniae, S.pneumoniae

 Capsulated organisms
Capsulated bacteria Composition
Pneumococcus Polysaccharide
Meningococcus Polysaccharide
Haemophilus influenzae Polysaccharide
Klebsiella pneumoniae Polysaccharide
Pseudomonas aeruginosa Polysaccharide
Bacteriodes fragilis Polysaccharide
Bacillus anthracis Polypeptide (glutamate)
Streptococcus pyogenes Hyaluronic acid
Capsulated fungus
Cryptococcus neoformans Polysaccharide
• Both capsule and slime layer as in Streptococcus salivarius.
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Additional Organelles
3. Flagella –
 Long (3 to 12 µm), filamentous cytoplasmic
appendages protruding from cell wall
 Organs of locomotion

 Chemically, composed of proteins called flagellins

 The number and distribution of flagella on the bacterial

surface are characteristic for a given species - hence are
useful in identifying and classifying bacteria
 Flagella may serve as antigenic determinants (e.g. the H
antigens of Gram-negative enteric bacteria)

 Presence shown by motility e.g. hanging drop preparation

Types of flagellar arrangement
Polar/ Monotrichous – single
flagellum at one pole- V. cholerae

Lophotrichous – tuft of flagella at one

Pole- spirilla

Amphitrichous – flagella at both

Poles. Alcaligenes fecalis.

Peritrichous – flagella all

over- salmonella
Demonstration of Flagella:
-0.02 µm in thickness beyond the resolution
limit of the light microscope.
1. Dark ground illumination
2. Electron microscopy
3. Special techniques
4. Indirect method
i. Swarming growth of Proteus
ii. Hanging drop preparation
iii. Craigie’s tube method
iv. Motility agar
Additional Organelles
4. Fimbriae/ Pili –
 Thin, hairlike appendages on the surface of many Gram-
negative bacteria

 0.1-1.0 µ long, and less than 10 nm thick- -

1. organs of adhesion (attachment)
allowing bacteria to colonize environmental surfaces or
cells and resist flushing
 2. helps in conjugation (process of transfer of DNA)
Made up of proteins called-- Pilin


 Pili can be of two types –

 Common pili – short & abundant
Sex pili - small number( 1-6 ), very long
Additional Organelles
5. Spores –

 Highly resistant resting stages

formed during adverse
environment (depletion of
nutrients)
 Formed inside the parent cell,
hence called Endospores

 Very resistant to heat, radiation

and drying and can remain
dormant for hundreds of years.

 Formed by bacteria like:

Clostridia,
Bacillus
The cycle of spore formation and germination
Shape & position of bacterial spore

Oval central

Spherical central Non bulging

Oval sub terminal

Oval sub terminal

Oval terminal Bulging

Spherical terminal

Free spore
 Demonstration of SPORES
1. Gram staining: Spore appear as an unstained refractile body within the cell.

2. Modified ZN staining: spore appear as Acid Fast (red color)

3. Schaeffer-Fulton method:
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