3/11/2024

Flagella, Fimbriae
And
Bacterial spore
Bivila
ila
Biv

Flagella
• Cytoplasmic appendages
protruding through cell wall
• Composed of a protein called
flagellin.
• Organ of locomotion
• Flagellar antigen- H or
Hauch antigen
• Length 5-20 µ m and
diameter 0.01-0.02 µ m
Bivila

1
 3/11/2024

Structure
• Consists of 3 parts
– Filament
– Hook
– Basal body

• Filament
– Only part that extend external to cell
– Connected to hook at cell surface
– Attached to basal body through hook
Bivila
ila
Biv

• Hook
– Embedded in cell envelope
– Connects basal body with filament

• Basal body
– Anchor flagella to cell wall and cell membrane
– Contains outer and inner rings by which it is
attached to cytoplasmic membrane

Bivila

2
 3/11/2024

Bivila
ila
Biv

Bivila

3
 3/11/2024

Functions
– Helps in motility of bacteria
– Antigenic in nature

Bivila
ila
Biv

Demonstration
• Direct methods
– Electron microscopy
– Phase contrast microscopy
– Light microscopy
• Silver staining

Bivila

4
 3/11/2024

Demonstration
• Indirect methods
– Spreading growth on a medium
e.g. swarming growth of Proteus
– Motility under microscope
e.g. hanging drop method
– Semi solid agar
– U tube and cragies tube

Bivila
ila
Biv

Bivila

5
 3/11/2024

Hanging drop motility

Bivila
ila
Biv

Fimbriae/pili
• Hair like appendages projecting from cell surface
• Found in gram negative bacteria
• More numerous and stiffer than flagella
• Antigenic
• Made of protein called pilin
• Unrelated to motility
• TYPES
– Common pili
– Sex or F (fertility )pili
– Col I /colicin pili
Bivila

6
 3/11/2024

Function
– Adhesion
– Transfer of genetic material
Demonstration
• Direct method
– Electron microscopy, phase contrast microscopy

• Indirect method
– Haem- agglutination
– Pellicle formation in liquid media
Bivila
ila
Biv

Bacterial spore
• Under conditions of limited supply of nutrition or
unfavourable environmental condition certain
bacteria forms highly resistant dehydrated resting
stage known as spores

• TYPES
– Endospore - inside cell
– Exospore - outside cell

• e.g. Bacillus, Clostridium

Bivila

7
 3/11/2024

Morphology of spore
1. CORE
– Spore protoplast
– Contain nucleus, enzymes etc
2. SPOREWALL
– Innermost layer surrounding spore
3. CORTEX
– Thickest layer

Bivila
ila
Biv

4. SPORE COAT
– Keratin like protein covering
5. EXOSPORIUM
– Thin and delicate lipoprotein membrane

Bivila

8
 3/11/2024

Bivila
ila
Biv

SPORULATION
• Process of spore formation in vegetative cell

1. Axial filament formation

– DNA replicates to 2 copies

2. Septum formation
– 2 copies of DNA moves to 2 compartments made
by plasma membrane septum
Bivila

9
 3/11/2024

3. Compartment formation
– Invagination of large spore so that small one
placed in it
– Large spore engulf small spore
– Large spore-sporangium
– Small spore-forespore
4. Cortex formation
– Peptidoglycan, Ca ions and dipicolinic acid laid
between inner and outer layers to form cortex

Bivila
ila
Biv

5. Spore coat formation

– Coating over cortex
6. Maturation
7. Lysis
– Release of free spores
– Germinates to form vegetative cell

Bivila

10
 3/11/2024

AXIAL FILAMENT
FORMATION

SEPTUM FORMATION

LYSIS
COMPARTMENT FORMATION

MATURATION

Bivila
CORTEX FORMATION
ila
Biv

Bivila

11
 3/11/2024

RESISTANCE
– Resistant to boiling, disinfectants and heating
– Due to
• High content of Ca ions
• Dipicolinic acid
• Low water content
• Thick cortex
• Spore coats
• Low enzymatic and metabolic activity

Bivila
ila
Biv

GERMINATION
– Process of conversion of spore into vegetative
cell
FUNCTIONS
• Tide over adverse condition
• Help in spreading infection
• Used as sterilisation control
– Bacillus stearothermophilus kept at autoclave
– Absence of spore after autoclaving indicates
proper sterilisation
Bivila

12
 3/11/2024

DEMONSTRATION
• Unstained preparation
– Electron microscopy and phase contrast
microscopy
• Stained preparations
– Gram’s staining
• Spore appear as unstained refractile body

Bivila
ila
Biv

– Modified Ziehl Neelsen staining

• Spore appear as red
• Instead of 20% H2SO4 decolorising agent 0.25-
0.5% is used

Bivila

13
 3/11/2024

– Special staining methods

• Schaeffler fulton method-spore green
• Dornev’s method -spore red

Bivila
ila
Biv

Expected questions
• 5 marks
– Bacterial spore
– Flagella

• 3 marks
– Fimbriae
– Types of arrangement of flagella
– Demonstration of flagella
Bivila

14