Living material is organized in unit and

microorganism were living form of

microscopical size and usually unicellular in
structure originally organization is
unsatisfied.
3
Bacterial Cell

4
 Unicellular
 Circular DNA
 No organelles
 1/10th the size of eukaryotic cells
 Flagella-long hair-like structure used for
movement
 Reproduce asexually –Binary Fission
 Size of bacteria:
 Bacteria are so small because of that their size is
measured in a micron (u )
 Generally cocci are about 1u in diameter and
bacilli are 2 to 10 u in length and 0.2 to 0.5 u in
width
 The limit of resolution with unaided eye is about
200 u because of that bacteria can be only
visualized under microscope.
 Shape of bacteria:
 Cocci in cluster – staphylococci
 Cocci in chain – streptococci
 Cocci in pair- diplococci
 Cocci in group of four – tetrad
 Cocci in group of eight – sarcina
• In the bacteria the outer layer or cell envelop or
bacteria consist of two things
(a) rigid cell wall
(b) underlying cytoplasmic membrane or
plasma membrane:
it includes granules, ribosomes,
mesosomes and circular DNA.
 Some bacteria in addition to possess aditional
structures such as gelatinous material which
cover it is called as capsule and when it is too
thin it is called as microcapsule.
 Flagella
 Pili
 Capsule
 Plasma Membrane
 Cytoplasm
 Cell Wall
 Lipopolysaccharides
 Teichoic Acids
 Inclusions
 Spores

Chapter 4
 Introduction:
most cell posses a cell envelop
consisting of cell wall and underlying
cytoplasmic membrane.
 Definition:
“the tough, rigid structure which
surrounds bacterial cell it is called as cell wall”
• Thickness: 10-20nm
• Weight: 20-25% of dry weight of bacterial cell
wall
Chemical structure of cell wall:
 Chemical structure of cell wall is made up of a Peptido-
glycan Polymer (amino acids + sugars)
 That structure is unique to all bacteria
 Sugars; NAG & NAM
 N-acetylglucosamine
 N-acetymuramic acid
 Amino acids cross link NAG & NAM

Chapter 4
 Made of peptidoglycan – a combination of protein and
polysaccharides
 It is present in both gram positive & negative bacteria.
 Some bacteria called Gram negative bacteria have an additional
layer of membrane that contains lipopolysaccharide
- this extra layer inhibits the uptake of antibiotics – protecting the
bacteria
cell wall
cell
cell membrane
membrane
lipopolysaccharide

cell wall Outer membrane

 Gram negative cell wall:
 Lipoprotein layer: it connects the peptidoglycan
to outer membrane

 Outer membrane : this contain certain proteins

that work as a target sites for antibiotics.

 Lipopolysaccharide : it consist endotoxins

( which are responsible for pyrogenecity, lethal
effect, tissue necrosis)
 Periplasmic space : it is the space between
inner & outer membrane which contains
important proteins and oligosaccharides.
Gram positive cell wall:
 Peptidoglycan layer : in gram positive bacteria
peptidoglycan layer is thicker( 15-25 nm ) than
gram negative bacteria ( 10-15 nm )

 Gram positive cell wall contains antigens such

as polysaccharide & proteins.
 Function
 1) protection of internal structure
 2)Gives shape to the cell
 3) confers rigidity & ductility
 4) role in division of bacteria
 5) offers resistance to harmful effect of
environment.
 6) contains receptor sites for antibiotics
 7)Provide attachment to complement
Gram positive Vs gram negative
Chapter 4
Chapter 4
 Difference Gram positive Gram negative

1. Thickness 15-25 nm 10-15 nm

2. variety of amino acid Few Several

3. aromatic & sulfar containing Absent Present

amino acid

4. Lipid Low 2-4 % High 15-20 %

5. Teichoid acid Present Absent

6. Periplasmic space Absent Present

7. result of enzyme digestion Protoplast Spheroplast

The type of cell wall is used by
doctors to help diagnose disease

The bacteria are stained with a

special stain called Gram stain
Absorb stain appear purple
Bacteria without the extra membrane,
appear purple. These are Gram positive
(Gram +) bacteria

bacteria with the extra membrane

appear pink. These are Gram negative Don’t absorb stain appear
( Gram -) bacteria pink
Chapter 4
 Definition:
“thin semipermiable membrane which lies
just beneath the cell wall that is called as cyto
plasmic membrane”
 The whole bacterial cytoplasm is bound
peripherally by very thin,elastic and
semipermiable cytoplasmic membrane also
known as cell membrane.
 It is 5-10nm in width
 Electron microscope shows the presence of three
layer constituting a unit membrane structure.
 Chemically the membrane consist of
phospholipid with small amount of protein.
Sterol is absent except in mycoplasma.
 DEMONSTRATION:
 The seperation of membrane from cell wall is
achived by readily in gram negative bacteria
when they are suspended in medium of high
osmotic tension.such phenomenom is called as
plasmolysis
 Electron microscope
Chapter 4
Chapter 4
 Transport:
(1) Active transport:
it is site of numerous enzymes (oxidase
polymerase, permease) involved in the active
transport of selective nutrients. It is
impermiable to macromolecule nd ionised
substances.
(2) Passive transport:
It is act as semipermiable membrane
through inward and outward passage of water
and passive trnsport of molecule lipid soluble
solutes take place by diffusion
(2) Concentration:
it is also concentration sugar, amino acids
and phosphate so that a 300-400 fold grandient
exists across osmotic barrier.
(3) Enzymatic function:
it also contain cytochrome oxidase, enzyme
of tricarboxylic acid cycle and polymerising
enzyme necessary for synthesis of cell wall
 “Bacterial cytoplasm is
suspension of organic and inorganic
solutes in viscous watery solution”
 It is not exhibiting protoplasmic streaming
and it lacks endoplasmic reticulum or
mitochondria. It contains ribosomes,
mesosomes, inclusion and vacuoles.
 All the organills which cytoplasm contains
is as follows.
 Ribosomes:
(1)
Ribosomes appear as small granules and pack
the whole cytoplasm. These are strung together
on strands of mRNA to form polymers.
the code of mRNA is translated in to peptides
sequence t this place.
The ribosomal particles become linked up and
travels along the mRNA strand.
Function:
Site for protein synthesis.
 Polysomes:
They are the group of ribosomes linked
together like beads of chain by messanger RNA
 Mesosomes:
They are vesicular, convulated or
multilaminted structures formed as invagination
of plasma membrane in cytoplasm .
They are more prominent in the gram positive
bacteria.
 Two types of mesosomes
(a) septal mesosomes:
It is attached to bacterial chrosome nd involved in
dna segregation and in formation of cross wall during cell
division.
(2) Lateral mesosomes:
They are at lateral side
Functions:
 They are site of respiratory enzymes

 Coordinate nuclear and cytoplasmic membrane division

during binary fission
 Responsible for compartmenting DNA at sporulation
 “ It is an outer covering of thick jelly like
material that surrounds the bacterial cell wall”
 Width: 0.2 micrometer
 Contains about 90% water and 2% solid
 The solid constitutes may be complex
polysccharide (pneumococcus klebsiella,
enterobacter) or polypeptides (anthrax bacillus)
or hyaluronic acid (streptococcus)
 Demonstration:
the capsule is best seen in pathological
specimens like pus, blood, sputum, and exudates
(1) By ordinary stain (gram or acid fast) capsule
can not be stained, it apper as halo arround the
stained bacterial body.
(2) In negative staining (india pink preparation),
capsule appears as clear halo around the
bacterium as link can not penetrate capsule.
 Immunological method: in that stained
with antiserum that swallowed by capsule
so it appear as swollen under microscope
so it is called as capsular awelling
reaction. E.g pneumococcus
 Capsulated organism:
s.pneumoniae,Bacillus anthracis,
C.perfringes, pneumobacillus,
H.influenzae
 Capsule serve as protective covering
against antibacterial substances such as
bacteriophase, phagocytes and enzymes
 Enhance bacterial virulance
 Capsular antigen is hapten in nature and
specific for bacteria.
 Definition
 “These are long, sinnous contractile
filamentous appendages known as f lagella”.
 Composed of a flagellin subunit.
 Usually sheathed (covered).
 Rotates by way of a basal body in the
bacterial cell.
 Unique to bacteria.
 These are organs of locomotion . ex:-Escherichia
coli salmonella, vibrio , pseudomonas, etc.
 The number of f lagella varies up to 10 to 20 per
cells according to species of bacteria.
 These are extremely thin (diameter)12 to 30 nm,
helical shaped structure of uniform diameter
throughout their length .
 these are 3 to 20 nm long.
 Each flagellum consist of hook & basal body. It
originates in a spherical body located just inside
cell wall.
43
 Composed of filament, hook, and basal body
 Flagellin protein (filament) is deposited in a helix
at the lengthening tip
 Base of filament inserts into hook
 Basal body anchors filament and hook to cell wall
by a rod and a series of either two or four rings of
integral proteins
 Filament capable of rotating 360º
 Arrangement/ types
 Monotrichous; 1 flagella
 Lophotrichous; tuft at one end
 Amphitrichous; both ends
 Peritrichous; all around bacteria

Chapter 4
Function
 It is responsible for bacterial motility.- Motility
may be observed microscopically or by
 detecting the spreading growth in semi solid agar
medium.

Demonstration –
 Dark ground microscopy.
 Special staining techniques in which their
thickness is increased by mordanting.
 Electron microscop.
 Hanging drop preparation.
 Definition
 “Fimbria are filamentous , short , thin , straight , hair
like appendage”.
 This is 0.1 to 1.5 µ long & less than 4 to 8 nm thick.
 They are also called as Pili.
 Fimbriae are seen only in some gram negative bacteria.
 Each bacterium may have 100 to 500 Fimbriae on all
over the body of bacteria.
 They project from cell surface as a straight filaments.
 They are best developed in freshly isolated strains & in
liquid culture.
 They are composed of protein known as pillin (molecular
weight 18000 Daltons).
Different forms of fimbria –
 i)common pili
 ii)F (fertility) pili
 iii) Col I (colicin)pili
DEMONSTRATION:-
 Electron microscop.
 Hem agglutination.
 Fimbriated bacteria form pellicle in liquid
media.

FUNCTION:-
 a) Organ of adhesion.
 b) Hem agglutination.
 c) They are antigenic.
 d) Agglutination & pellicle formation.
 e) Genetic material is transferred from the
donor to recipient cell.
Sr.n Flagella Fimbriae
o

1 Size larger & thicker Smaller & thinner

2 Arise from cytoplasm or cytoplasmic membrane but Attached to cell wall

does not attached to cell wall

3 Organ of movement (locomotion) Organ of adhesion &

conjugation

4 They are never straight They are alwase strait

 “ spores are highly resistant dormant stage of
bacteria formed in unfavourble environmental
condition such as starvation and dessication”
 As spores are formed within the parent bacterial
cell so they are also called as endospores
 During germination each spore give rise to only
one vegetative bacteria
 Exospores found in fungi(conidia) formed
extracellularly from end of parent cells.
 Sporulation is not a method of reproduction
A. Gram positive bacilli:
(1) obliterate aerobic- genus bacillus.e.g
B.anthracis, B.subtilis
(2) obliterate anaerobic:genus clostridia.e.g
C.tetani, C.welchii, C.botulism
B. Other bacteria:
Gram positive coccus (porosarcina) gram
negative bacilli (coxiella burnetii)
 The cell membrane grows inward and forms
spore wall around the core (forespore). The
inner-most layer of the spore wall forms the
spore membrane from which the cell wall of
future vegetative bacterium develops. Outside
this membrane is thick layer, the cortex and a
multilayered tough spore coat. Some spores
have an additional apparently rather loose,
outercovering called exosporium.
 spontaneous sporulation occurs in
condition unfavourable condition such as
starvation, dessication, presence of
disinfectants and in extreme temperature.
http://www.microbe.org/art/endospore_cycle.jpg 58
Spore formation is initiated by apperance of clear area in
portion of protoplasm near one end of bacterial cell and
that protoplasm gradually become more opaque that form
forespore

The cell membrane grow inwards and undergoes infolding

forming double layered membrane structure around the core

The inner most layer of spore wall forms spore membrane in

future vegetative bacterium will develop. The spore wall
synthesis a thick covering lyer cortex and multilayered thin
but tough outer layer spore coat
Spores of some spesis have additional apparently
rather loose outer covering called exosporium

Spore cortex contains unusual type of peptidoglycan

sensetive to lysosomes the spore cot is made up of keratin
which is impervious to antibacterial chemical agent

Exosporium is a lipoprotein membrane with some

carbohydrate residue. Young spores remain attached to
parent cell
 The young spore remain attached t parant
cell. The precise position and shape and
relative size of spores remain constant within
particular bacteria.
 Spore may be central,sub terminal or
terminal
 Bacterial spores are resistant to ordinary boiling,
heating, and disinfectant. They can withstand
boiling up to 3 hr, dry heat at 150c for 1 hr
however they are destroyed by autoclaving at
121c for 15-20 min.
 The highly impervious spore coat, low water
content, low metabolic activity and high
concentration of calcium dipicolinate of spore
make resistant to drying and heat.
 Definition:
“ The process of conversation of spore in
to vegetative cell under suitable environment is
known as germination”

There are three stages of germination

 The germintion of bacterial spore do not
occur even when placed in environment that
favour process.
 Unless first activated by one or onother
agent damage the coat of spore such as heat,
abration and compound containing free
sulphydryl groups.
 The process of initiation is not clear, however
the spore will initiate germination in
favourable condition.
 Different species of bacteria recognizes
different effactors as signalling a rich
medium such as L-alanine for one spesis
 With the swelling of spore wall and
disintegration of cortex a single germ cell
emerge after breaking open the spore coat.
 The new vegetative cell consist of spore
protoplast with its surrounding wall.
 Formation of vegetative cell.
 By ordinary stain and modified Z-N stain.
 Laboratory:
for making sterilization
Chapter 4
 Pili – hairlike structures usually found
in Gram neg. bacteria. Help the
bacteria stick to surfaces.
Also forms conjugation bridge
 Chromosome – a single loop of DNA
that is folded on itself
- controls the cell’s function
 Nucleoid – the region of the cytoplasm
where the DNA is found
 Plasmid – an accessory loop of DNA – small contains only a few
genes - can be responsible for: conjugation, antibiotic
resistance, unique metabolic properties – like the ability
to use hydrocarbons
 Capsule – found outside some bacteria stores nutrients and protects
the bacteria from changing environmental conditions
 Short protein appendages
 smaller than flagella
 Adhere bacteria to surfaces
 E. coli has numerous types
 K88, K99, F41, etc.
 Antibodies to will block adherance
 F-pilus; used in conjugation
 Exchange of genetic information
 Flotation; increase boyancy
 Pellicle
(scum on water)
 More oxygen on surface

Chapter 4
Chapter 4