Morpholofy of Mo

MORPHOLOGY
OF
MICROORGANISMS
CLASSIFICATION SYSTEM
Bergey's Manual
of Determinative Bacteriology the
"bible" of bacterial taxonomy.
There are such levels of microorganisms
organization:
Species Genus Family Class
Division Kingdom
Bacteria are distinguished primarily on:

a) morphological characteristics cell shapes
(rods, cocci, curved, or filament
forming)
b) spore production
c) staining reactions
d) motility
)Other groups are defined based on their
metabolism, or combinations of
morphological and physiological
characteristics.
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
BACTERIA WITH SPORES

Oval central
Spherical central
Oval sub terminal
Oval sub terminal
Oval terminal
Spherical terminal
Free spore
Bulging
BACTERIA WITH FLAGELLA

Polar/ Monotrichous single
flagellum at one pole
Lophotrichous flagella at one
pole
Amphitrichous flagella at
both poles
Peritrichous flagella all
over
Amphilophotrichous flagella
at both ends
STAINING OF BACTERIA
Bacteria cells are almost colorless
and transparent
A staining technique is often applied
to the cells to color them

Their shape and size can be easily
determined under the microscope.
PRINCIPLE OF STAINING
Stains combine chemically with the
bacterial protoplasm.
Commonly used stains are salts:
Basic dyes: colored cation + colorless anion
e.g : methylene blue (methylene blue
chloride)
MB+ + Cl Acidic dyes: colored anion + colorless
cation
e.g : eosin ( Na+ + eosin-).
TYPES OF STAINING
Simple stains
Differential stains
Gram stain
Acid-fast stain
Endospore stain
Special stains
Negative (capsule) stain
Flagellar stain
Fluorescent stains
Staining for electron microscopy
TYPES OF STAINING
Simple staining
(use of a single stain)

For visualization of
morphological
shape & arrangement
Differential staining
(use of two contrasting stains

separated by a decolorizing agent)
Identification
Gram
stain
Acid fast
stain
Visualization
of structure
Spore
stain
Capsule
stain
SIMPLE STAIN
Simple stains provide a quick and easy way to
determine cell shape, size, and arrangement.

1) Perform a bacterial smear.
2) Saturate the smear with basic dye for
approximately 1 minute. You may use
crystal violet, safranin, or methylene
blue.
3) Rinse the slide gently with water.
4) Carefully blot dry with bibulous paper.
5) Observe the slide under the microscope,
using proper microscope technique.
BACTERIAL SMEAR
BASIC SHAPES
Cocci
Bacilli
GRAM STAIN
With Gram stain, bacteria could be divided
into:
Gram positive bacteria: bacteria that
retain crystal violet iodine dye complex
and so appear purple.
Gram negative bacteria: bacteria that
destain with 95% alcohol and appear red
due to counterstaining with carbol fuchsin.
This difference in staining affinity is due to
difference in the permeability of cell wall.
Interpretation:
1.
2.
Gram positive: dark blue / violet

Gram negative : pink / red
CHAINS OF COCCI
Streptococcus pyogenes
FUNGI IN GRAM STAIN
CLUSTER OF COCCI
Staphylococcus aureus
Sputum smear
BACILLUS SHAPED BACTERIA
Escherichia coli
Pseudomonas aeruginosa
Fusobacterium
BACILLUS CHAIN
Streptobacillus
moniliformis
PALLISADES ARRANGEMENT
Corynebacterium diphtheriae
CURVED / COMMA SHAPE
Vibrio cholerea
SPHIROCHETES
Sphirochetes
Treponema pallidum
ZIEHL NEELSEN STAIN

This stain is used for detection of acid
fast bacteria.
These bacteria are not stained with
ordinary stains but they need exposure to
strong stains with application of heat.
Once stained, they will resist
decolorization with mineral acids such as
H2SO4 or HCL.
This property is due to large amount of
lipids and fatty acids especially mycolic
acid wax in cell wall of these bacteria.
PROCEDURES
Interpretation:
1. acid-fast bacteria: red
2.
non-acid fast bacteria: blue
Mycobacterium spp.
BURRY-GINZS STAIN
Objective: Determine morphology
and arrangement
India ink used to stain background,
not cells
Gives a good view of morphology
Not heat fixed, so cells are not
distorted.
ENDOSPORE STAIN
Oscheshko stain
1) Thick layer of sporulating bacteria smeared
on slide.
2) Dried without fixation in normal room
condition.
3) Pour 0.5% HCl for 2mins with heating.
4) Wash and dry.
5) Fixate smear on burner.
6) Next stain as per in Ziehl Neelsen stain.
Schaeffer-Fulton endospore stain

1. Endospore walls are very resistant to
penetration of ordinary stains
2. Malachite green: primary stain- heated
until steam to make endospore walls more
permeable to dye
3. Washing with water will remove the green
dye from all parts of the cell except the
endospore
4. Safranin: counterstain- stain the nonspore-forming or vegetative cells red.
Genera of bacteria that produce
endospores
1. Bacillus & Clostridium
2.
Other genera:
) Desulfotomaculum
) Sporosarcina
) Sporolactobacillus
) Oscillospira
) Thermoactinomyces
Interpretation:
Spores: green
Vegetative cells: red
ROMANOWSKY-GIEMSA STAIN
To observe bacterial nucleoid

1. Dried smear of pure culture prepared
(B.cereus).
2. Fixate in Karnua Solution,
3. Applicate HCl with heating.
4. Stain the smear with Romanowsky-Giemsa
dye (azur, eosin, and methlyene blue) for
40-60mins.
5. Wash with water and dry the stained slide.
FLAGELLA STAIN
Purpose: To determine the presence
/absence and location of flagella on various
microorganisms.
Principle: Because bacterial flagella are
very thin and fragile a special stain
(flagella stain) is prepared that contains a
mordant. This mordant allows piling of the
stain on the flagella, increasing the
thickness until they become visible. Various
arrangements of flagella are seen on
different cells.
1. Add about 1 ml. of the Flagella Stain

solution (one dropper full) to the smear and
allow to stain for 10-15 minutes. The
solution should not be allowed to flow outside
the waxed area.
2. Flood off the stain by adding tap water to
the slide while it remains on the rack. Do
not tip the slide before this is done.
3. Drain and flood the slide with carbol fuchsin
for one minute. Rinse by flooding. Drain and
air dry. Do not blot.
Bacillus brevis
Proteus - Peritrichous Arrangement of

Flagella
THE END
THANK YOU
PREPARED BY: PATHUMAVATHY RAMANTHAN (21B)

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MORPHOLOGY

Bacteria are distinguished primarily on:

BACTERIA WITH SPORES

BACTERIA WITH FLAGELLA

A staining technique is often applied

to the cells to color them

Staining for electron microscopy

(use of a single stain)

(use of two contrasting stains

determine cell shape, size, and arrangement.

Gram positive: dark blue / violet

FUNGI IN GRAM STAIN

BACILLUS SHAPED BACTERIA

CURVED / COMMA SHAPE

ZIEHL NEELSEN STAIN

non-acid fast bacteria: blue

Schaeffer-Fulton endospore stain

Genera of bacteria that produce

To observe bacterial nucleoid

1. Add about 1 ml. of the Flagella Stain

Proteus - Peritrichous Arrangement of

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