BACTERIAL CELL

ULTRASTRUCTURES
General Division:

I. External - Capsule
- Fimbriae
- Flagellum

II. Internal - Cell Wall

- Cytoplasmic Membrane
- Cytoplasm
- Mesosome
- Nuclear Body
- Ribosome
- Plasmid
- Inclusion Granule
- Endospore
 BACTERIAL ULTRASTRUCTURE

A. Bacterial Surface Coating:

= are viscous extracellular polymers which surrounds
the bacterial cell

Capsule
= well-formed thick viscous jelly-like structure firmly
attached to the cell wall surrounding the cell
= not readily removed.
= easily visualized by negative staining using India
ink method
Slime layer

= loose and irregularly arranged network of

fibrils totally detached from the cell
extending outside the cell surfaces

= when present are easily washed off

Chemical Comp.:
= water – main component 98-99%
= 1-2% of bacterial capsule are chemically
polysaccharide, Except capsule of:
B. anthracis-polypeptide (D-glutamic acid)
S. pyogenes-Hyaluronic acid
Function:
= protection from phagocytosis
= correlates with virulence
= act as antigen (used in the identification)
and typing of capsulated bacteria
= allows bacteria to adhere/attach to various
surfaces in its natural environment in
order to survive
Detection:
1. Negative staining (India Ink)
2. Special staining (Capsular method)
3. Serological (Quellung reaction)
B. Appendages Flagellum & Axial filaments
Fimbriae

FLAGELLUM:
= long, thick, helical protein filament of uniform length
and diameter
= commonly seen among free-swimming bacteria
= originates in cytoplasmic membrane
= composed of 3 parts:
1. Basal body – anchors the flagellum to the
cell wall and plasma membrane

2. Hook – attached to the basal body and

connects to the filament

3. Filament – external to the cell and contains

the flagellar protein
Chemical Comp.: protein monomer - Flagellin

Function - Motility organelle of many pathogenic bacteria

Detection:
1. Darkfield / Phase contrast microscopy
(Wet mount / Unstained smear)

2. Brightfield – using stained specimen

3. Electron Microscopy – stained specimen

4. Serological – use of specific antisera against

flagellar (H) antigen
Types and Arrangement of flagella in relation to the
distribution and number:

I. Monotrichous – single polar flagellum at one end

II. Multitrichous – more than one flagella

Types:
A) Lophotrichous – tuft of flagella at one polar end

B) Amphitrichous – single flagellum or tulf of

flagella at both polar end

C) Peritrichous – flagella distributed all over the

body of the bacteria
 Axial Filaments
= flagella-like sheathed filaments located in
the periplasmic space (between the inner)
and outer membrane of the cell
= move by traveling helical wave on opposite
direction
= motility organelle of spirochete

Fimbriae / Pili / Microfibrils

= short, straight, thin hair-like filaments usually
distributed around the body of bacteria.
= originates in the cytoplasmic membrane
= found virtually among all gram (-) bacteria but not in
gram (+) bacteria
2 types according to function:

1. Common/ordinary – for attachment or adherence

to mucosal surfaces of host cell during
colonization and infection.

2. Sex pili – responsible for attachment of donor and

recipient cell during bacterial conjugation
whereby genetic material (DNA) from one cell
is transferred to another (reproduction).

Chem. Comp. - Protein (Pilin)

Detection: Electron Microscopy

Cell wall (Peptidoglycan, Murien, Mucopeptide / Glycopeptide)
= complex, rigid, multilayered structure that protects the
protoplast and the underlying fragile plasma
membrane

= found in all pathogenic free-living bacteria except

Mycoplasma (cell-walless bacteria)

Chemical Comp.:
1. Protein (Mesodiaminopimelic acid,Isomers of D-glutamic)
acid and D-alanine

2. Polysaccharide
(N-acetylmuramic acid and N-acetylglucosamine)
= responsible for rigidity of cell wall

3. Lipids
Function :
1. Responsible for the characteristic shape of bacterium
2. Provides strong structural support necessary to keep
bacterial cell from rupturing due to changes in
the environmental osmotic pressure
3. Contain somatic O antigen that can serologically
identify particular bacteria
4. Site of action of some antibiotics
5. Determines differences in gram staining reaction

Detection :
1. Microscopy of smear prepared from special
staining method.
2. Electron microscopy.
3. By chemical methods using lysozyme.
 COMPARISON OF GRAM POSITIVE AND GRAM NEGATIVE CELL WALL

Gram Positive Gram Negative

Peptidoglycan Verythick homogenous single layer
extensively crossed-linked.
Constitute 50-90% of the cell wall
material.
(NAM, NAG, D-alanine)
Diaminophemilic acid
More complex layer composed
of thin peptidoglycan with few
crossed-linked and an outer
membrane layer.
Constitute 5-10% of the cell
wall material.

Auxiliary compounds Teichoic acid (Polymer of ribitol and) No teichoic acid
glycerol phosphate Outer membrane contains:
Lipoteichoic acid Phospholipids-located in the
Surface protein inner membrane
Lipoprotien – connects OM to
the peptidoglycan
Lipopolysaccharide
(LPS/Endotoxin layer)
– located in the outer
layer of the OM
- contains the lipid A
Penicillin sensitivity Sensitive Resistant

Response to Digest/ Degrade Resistant

lysozyme
Gram
Positive

Gram
Negative
Damage to the cell wall may result to:

Protoplast
= when a gram positive bacteria is exposed to
lysosyme will degrade the peptidoglycan
layer resulting to complete removal of the cell
wall producing a wall-less spherical body
= since all cell wall components is removed,
therefore incapable of regeneration

Spheroplast
= when a gram negative bacteria is exposed to
lysozyme it looses the peptidoglycan layer
but retain the outer membrane leaving a
less fragile spherical body which are capable
of regenerating the cell wall.
Protoplast: comprises the naked cytoplasmic membrane and
its content

PERIPLASM = space between the plasma inner membrane

and the outer membrane layer
= readily observe among gram negative bacteria
(with difficulty among gram + bacteria)
= consist of a gell-like substances which help
secure nutrients from the environment

CYTOPLASMIC/PLASMA/CELL MEMBRANE:
= thin elastic bilayered semi-permeable membrane
lying underneath the cell wall enclosing the
cytoplasm of the cell.

Chemical Composition - Lipoprotein -

(composed of phospholipids and protein
molecules)
Functions:
1. Serve as selective permeability barrier
(transport of nutrient material in & out of cell)
2. Site of electron transport and oxidative
phosphorylation of substances involve in the
generation of chemical energy (ATP)
3. Helps in the excretion of metabolic waste product
4. Site for excretion of enzyme involved in OM
synthesis, CW synthesis and in the secretion
of extracytoplasmic & extracellular substances:
A. Permease for active uptake of nutrient
B. Respiratory
C. Polymerase that manufacture substances
of the cell wall
D. Hydrolytic

Detection - Electron microscopy

Cytoplasm = refers to everything that is enclosed by the
cytoplasmic membrane
= site for most bacterial metabolism
= 80% of cytoplasm is composed of water
MESOSOME:
= usually seen as an invagination of the membrane
associated cytoplasmic sac seen in Gram (+) cell
= contains lamellar, tubular and vesicular structure
= often associated with division septa
Chemical Composition - Lipoprotein
Functions:
1. Site for the synthesis of cytochrome oxidase
and reductase enzyme
2. Provide support for enzymes present in the cell
membrane
3. Responsible for compartmenting DNA during
cell division and sporulation
Detection- Electron microscopy
NUCLEAR BODY: (Nucleus / Nucleoid)
= genetic material of bacteria
= contains a single circular molecule of double
stranded DNA network which runs parallel to the
long axis of the cell
= found homogenous within the cytoplasm not
enclosed in a nuclear membrane.
= constitute 2-3% of the cell weight.
= no definite form and seen at all stages of growth
cycle of the bacteria.
Chemical Composition – DNA
Function:
1. Controls the growth and metabolic activity of
the bacteria
2. Responsible for hereditary characteristics of
the cell.
Detection- Electron microscopy, Feulgin staining (+)
PLASMID/EPISOME:
= extrachromosomal genetic material capable of
autonomous replication usually located near
the chromatin body.

Chemical Composition – DNA

Function:
1. Transfer of genetic material from one cell to another
by conjugation (Transmissible Plasmid)
2. Carry genes for activities like:
a) Antibiotic resistance
b) Toxin production
c) Synthesis of enzyme
d) Tolerance to toxic metals

Detection– Feulgin staining (+), Electron microscopy

RIBOSOME:
= histone-like particles composed of ribosomal RNA
(rRNA) and protein molecules found in the
cytoplasm of the cell.
= site of action for many antibiotics that inhibit protein
synthesis.
= made up of 2 subunits
larger subunit (50S)
smaller subunit (30S)
= size 10-15nm with sedimentation coefficient of 70S
= procaryotic ribosome - 70s
= eucaryotic ribosome - 80s

Function – Site of protein synthesis

Detection– Feulgin staining (-), Electron microscopy

INCLUSION / CYTOPLASMIC GRANULE:
= large granules found anywhere in the cytoplasm
= are accumulation of organic and inorganic substances
which serves as source of energy and nutrient
supply to the organism

Kinds:
1. Babes Ernst / Volutin / Metachromatic granules

Chemical Composition – polymerized

metaphosphate which can be used in the
synthesis of ATP

Function – reserve energy supply

Demonstration – simple staining / special

staining (Albert’s stain)
2. Lipid granules

Chemical Composition – polymer of beta-

hydroxybutyric acid
Function – reserve food supply.

3. Sulfur granules – derived energy by oxidizing

sulfur and sulfur-containing
compounds.

Function – reserve energy supply for sulfur-

containing bacteria.

4. Glycogen granule – polymer of glucose.

Function – reserve food supply.

ENDOSPORES:

= highly refractile body formed within vegetative

bacterial cells in response to adverse environmental
condition and due to deficient nutrient supply
= known as endospore, because it is formed inside the
bacteria cell
= formation of endospore is genetically controlled
= another life of the cell in a resting phase
= found in the cytoplasm of rod-shaped sporeforming
bacteria of the genus Bacillus & Clostridium
Classification accdg. to:

A. Location - Central, Subterminal, Terminal

B. Shape - a) Ovoid
b) Circular

C. Swollen/not swollen
(swollen when the diameter is more than
the of the bacterial cell, not swollen
diameter the same as bacterial cell)
Composed of five parts:
1. Core – located at the center which is the spore
cytoplasm/protoplast.
- contains the nucleus and enzyme dipicolinic acid
2. Spore wall – enclosing the core.
3. Cortex – laminated structure surrounding the spore
wall
4. Spore coat – multilayered membrane enclosing the
cortex
5. Exosporium – the outermost covering of the spores
which give the spore a rigid appearance.
Chemical Comp. – Calcium Dipicolinate

Function: serves as another life of the cell (survival)

= can undergo spore formation or sporulation
= only one spore is found in a bacterial cell

Detection:
1. Microscopy of stained smear
A) Negative staining
B) Positive staining

2. Phase contrast microscopy