BACTERIA

STRUCTURE
NUTRITION categories
RESPIRATION categories
CLASSIFICATION/IDENTIFICATION (Gram stain)
REPRODUCTION and GROWTH
MICROBIOLOGY TECHNIQUES
• Medium, sterile technique, staining, microscopy,
inoculation and isolation of pure culture
techniques
 STRUCTURE
CELL STRUCTURE; SIZE SHAPE
ARRANGEMENT, WALL
STRUCTURE, CAPSULE
STRUCTURE, FLAGELLA
 Prokaryote vs Eukaryote
• https://www.youtube.com/watch?v=zZtcM
BTQaS4

• Make you own notes – prokaryote

structure, compare with eukaryote cell
structure: similarities and differences
 What you should know
• Structure- comprise the components of the
unicellular structure.
• You should know their location shape size
function within and on the cell.
• Structure also refers to size of the cells,
the shape of the cells and the
arrangement of the cells.
Bacterial prokaryote cell structure

Cell wall +
plasma or cell
membrane =
cell envelope
Some bacteria have additional
structures
 Size of bacterial cells

• Average size 1.3 x 4.0μm

• Nanobacteria - 0.2 – 0.05 μm in diameter

• Atypically large bacteria eg.Epulopiscium

fishelsi - 80 x 600μm
 SHAPE AND ARRANGEMENT
Name and describe all shapes and arrangements in the
image below
 Prokaryote structure: Cell wall
• The composition of the cell wall differs
significantly between the domains Bacteria
and Archaea.

• The composition of their cell walls also differs

from the eukaryotic cell walls found in plants
(cellulose) or fungi and insects (chitin).

• The cell wall functions as a protective layer,

and it is responsible for the organism’s
shape.
 Prokaryotic structure: capsule
• Some bacteria have an
outer capsule outside the cell wall.
• Not present in all cells
• Function of the capsule:.
- the capsule found in some species
enables the organism to attach to surfaces
- protects it from dehydration
- and attack by phagocytic cells,
- and makes pathogens more resistant to
Cell Envelope – capsules and
slime layers
• Function of the cell envelope:

– Part of pathogenicity mechanism of the bacterium

– Prevents phagocytosis by other microorganisms

– Helps the cells adhere to surfaces in aquatic

environments

– Prevent desiccation and death of the cell in dry

conditions
 Prokaryotic structure
• Some species also have flagella (singular,
flagellum) used for locomotion,

• pili (singular, pilus) used for attachment to

surfaces.

• Plasmids, which consist of extra-

chromosomal DNA, are also present in
many species of bacteria and archaea.
 Cell envelope
• The cell envelope = cell or plasma
membrane + cell wall
• The cell envelope of some bacteria may
also include: capsule or slime layer
 Cell membrane

Diagram of the macromolecules which make up the phospholipid

bilayer. If the membrane is broken the bacteria dies.
 Cell envelope - cell wall
This structure is also porous. Small molecules
are able to pass through the cell wall to the
membrane, but large molecules are not. The
cell wall acts as a coarse filter.

Nearly every genus of bacteria has a cell

wall, which is a rigid, carbohydrate-
containing structure that surrounds the
bacterial cell.
Gram – negative bacteria consists of a thin layer of peptidoglycan in the
periplasmic space between the inner and outer (2) lipid membranes,
the outer membrane consists of lipopolysaccharide.

b. Gram – positive bacteria have a single lipid membrane surrounded by

a cell wall composed of a thick layer of peptidoglycan.

Nature Reviews/ Microbiology

http://www.nature.com/nrmicro/journal/v13/n10/nrmicro3480/metrics
 Capsule/glycocalyx
• Capsule – compact layer of polysaccharide
surrounding the bacteria
• Slime layer – loose layer non-uniform in
density and thickness, tangled
polysaccharides
 Cell envelope – capsule
• Capsule present outside cell wall in some
bacteria

• Capsule :
– attachment to surfaces
– prevents desiccation in dry conditions
– resist immune response of humans
 Cell envelope - Flagella and
pilus

• The flagella allow the bacteria to be

motile and the pilus facilitates the
attachment of the bacteria cells to
surfaces
FLAGELLA
 a b

Tubes a and b are cultures of different species of bacteria

Which tube a or b have bacteria cells with flagella as part of
the structure?
 MICROBIOLOGY

TECHNIQUES USED FOR

ISOLATION,GROWTH, AND
CLASSIFICATION OF BACTERIA
 Microbiology
To study bacteria microbiologists culture
(grow) bacteria in the lab on artificial
medium under controlled environmental
conditions.
Bacteria which grow in the lab are referred
to as being culturable.
Only 5 % of bacteria can be cultured in the
lab
Artificial media /Bacteria culture
(Plates)
http://www.microbiologyinpictures.com/bacteria-photos/escherichia-coli-
photos/e.coli-on-agar.jpg
 Mixed culture vs pure culture

http://upload.wikimedia.org/wikipedia/com http://iws2.collin.edu/dcain/4way.h1.jpg
mons/f/fe/VCU_agar_plate_colonies.jpg
Nutrient agar can support growth of many species cuturable
bacteria. This is the artificial medium which will be used for our lab
exercises.
Culture of bacteria
Simple stain vs negative stain
/direct vs indirect

http://microscopystaining101.weebly.com/
Bacteria can be classified based
on the Gram’s stain
• This differential staining procedure of the
Gram stain separates most bacteria into
two groups on the basis of cell wall
ultrastructure (macromolecule
composition). The Gram positive bacteria
have a thicker layer of peptidoglycan and
stains purple, the Gram negative bacteria
have thin peptidoglycan layers and high
lipid content in the cell wall and stains
red/pink
• The Gram positive bacteria have a thicker
layer of peptidoglycan and stains purple,
the

• Gram negative bacteria have thin

peptidoglycan layers and high lipid content
in the cell wall and stains red/pink
Identification – Gram stain
procedure

1.Unstained bacteria

2.crystal violet (CV) stained

bacteria

3.iodine – CV coloured complex.

4.decolourise or remove iodine -

CV complex

5.Counter staining with

safranin
Gram stain
The thick layer of peptidoglycan in the cell wall
of Gram positive bacteria traps more crystal
violet – iodine complex, therefore the alcohol
step does not decolourise (remove) all of the
colour complex; so the safranin stain is
overshadowed by the coloured complex giving
the bacteria a purple colour. Alternatively, the
Gram negative bacteria have very little colour
complex remaining after the decolourisation
step as the colour complex is removed due the
removal of the lipid layer by the organic solvent
and release of the complex from the thin
peptidoglycan layers. The Gram negative
bacteria thus appears pink.
 Endospore stain

http://www.austincc.edu/microbugz/images/
endosporestain.jpg
STAINING
 CLASSIFICATION

Based on oxygen requirements

Aerobic and anaerobic
catabolic pathways
Five classes of bacteria based on their
requirement for oxygen

Obligate Facultative Aerotolerant Strict

Microaerophile
aerobe anaerobe anaerobe anaearobe
CLASSIFICATION

Based on TEMPERATURE
Effect of temperature on
bacterial growth
Bacterial classification based on
temperature
 Bacterial categories based on
temperature
• Bacteria group based on temperature
Temperature range (Celcius)
• Psychrophiles -5 -20
• Mesophiles 10-50
• Thermophiles 40-75
• Hyperthermophiles 65-95
 CLASSIFICATION

NUTRITIONAL CATEGORIES
 Nutritional categories
Nutritional category Energy source Carbon source

Photoautrophs Light CO2

(some bacteria and eukaryotes)

Photoheterotrophs Light Organic compounds

(some bacteria)

Chemoautotrophs Inorganic compounds CO2

(some bacteria and many archaea)

Chemoheterotrophs Organic or sometimes inorganic Organic compounds

Most known bacteria and archaea compounds
are chemoheterotrophs, also
fungi and protists
REPRODUCTION AND
GROWTH
Binary fission growth curve,
generation time
 Reproduction – Binary fission

https://sites.google.com/site/sacebiologystage1/home/reproduction/binary-
fission
 Growth rate curve – the rate
of growth

• What’s on the y axis (no. of cells, optical

density (od), no. of colony forming units
(cfu), log of the number of cells

• What’s on the x axis – time (hours or

days)
 Reproduction = Growth

Growth curve of a bacterial culture

 Generation time
Generation
time – the
time it takes
for the cell
number to
double in
number .

http://wswx.taru.edu.cn/source/whole/image/6/6.4.jpg