URUK UNIVERSITY

PRACTICAL MICROBIOLOGY LAB

THIRD GRADE

LAB 7

Bacterial Identification

Laboratory diagnosis

Infections can be diagnosed either directly by detection of the

pathogens or their components or indirectly by antibody detection
methods.

Identification of pathogen is based on morphological, physiological and

chemical characteristics. Bacterial identification was based on:

1. Macroscopical characteristic (Colonial morphology of bacterial cells).

2. Microscopical examination (Morphology of bacterial cells).
3. Biochemical tests.
4. Antibiotic sensitivity test.
5. Serological tests (Antigen and antibody detection test).
6. Nucleic acid assays.
7. Animal pathogenicity.

1. Macroscopical characteristics:
A. Colony morphology: colony size, shape and color.
B. Culture characteristics: as lactose fermentation, type of
haemolysis, swarming phenomena, pigment production.

Characteristics

These are the characteristics used to accurately and consistently

describe the morphology of a bacterial colony:

Edge (margin)
1 by Dr.Sana A.Alshakely
M.Sc. Samar M.M.
 URUK UNIVERSITY
PRACTICAL MICROBIOLOGY LAB
THIRD GRADE
Table : Colony Characteristics

No. Colony characteristics Observations

1 Size
2 Form
3 Elevation
4 Margin
5 Color
6 Haemolysis
7 Pigment production
8 Oder
9 Opacity
10 Surface
11 Consistency
Very small, Small, Medium, Large, Very
large.
Punctiform, Circular, Filamentous,
Irregular rhizoid, Spindle.
Flat, Raised, Convex, Pulvinate,
Umbonate.
Entire, Undulate, Lobate, Erose,
Filamentous, Curied.
White, Grey, Yellow, Black, Orange,
Pink, Red, etc
Haemolysis α, β or γ
Color of the pigment production.
Fruity, Freshly cut apple, Fishy, Fecal or
putrid, Bleach, Pungent.
Transparent, Opaque, Translucent.
Smooth, Glistening, Rough, Dull.
Buttery, Viscid, Brittle, Mucoid.

2. Microscopical examination (cell morphology)

Depended on the morphology of stained slide of bacterial cells

prepared from pure culture to determine either shape,
arrangement, gram reaction number and arrangement of flagella
if they are present, presence or absence of capsule and presence
or absence of spores.
1. Gram reaction: microscopic examination is usually done by
way of the gram stain procedure. This staining method will
differentiate gram-positive (purple) bacteria from gram-
negative (pink) bacteria. This is based on the composition of
2 by Dr.Sana A.Alshakely
M.Sc. Samar M.M.
 URUK UNIVERSITY
PRACTICAL MICROBIOLOGY LAB
THIRD GRADE
the cell wall of the organism. Common gram-positive bacteria
are staphylococcus and streptococcus. Gram-negative bacteria
include E. coli and Klebsiella.
2. Size of bacterial cell in micrometer.
3. Shape of bacterial cell.

Coccus= spherical
Bacillus
Coccobacilli= elongated coccal from
Spirillum= spiral
Vibrios= short, slightly curved rods
Fusiform=bacilli with tapered ends
Spirochete

4.Aggregation of bacterial cells: Bacteria sometimes occur in

groups, rather than singly, and the single cell's shape
influences the cell arrangements that they form as the
bacterial cells divide.

Bacilli divide along a single axis, and are sometime seen in pairs
or chains. Since they only divide along one axis, you will not
find bacilli in clusters, such as those formed by Staphylococcal
bacteria.
Cocci divide on one or more planes, producing cells in:
3 by Dr.Sana A.Alshakely
M.Sc. Samar M.M.
 URUK UNIVERSITY
PRACTICAL MICROBIOLOGY LAB
THIRD GRADE
Pairs (diplococcic)
Chains (streptococci)
Packets (sarcinae)
Clusters (staphylococci)

5. Arrangement of flagella on bacterial cells:

Bacterial flagella are thread-like appendages composed
entirely of protein, 12-30 nm in diameter, they are the organs
of locomotion for the form that possess them, the types of
arrangement are:

4 by Dr.Sana A.Alshakely
M.Sc. Samar M.M.
 URUK UNIVERSITY
PRACTICAL MICROBIOLOGY LAB
THIRD GRADE

6. Spore shape and location of spore within the bacterial cell:

during growth, some bacteria form a highly resistant structure
called an endospore. The endospore is formed within a
vegetative cell by a process called sporogenesis and is released
upon death of the vegetative cell.
Bacterial spore have either spherical or oval shape with a
central, terminal or subterminal position.

7. Capsule formation: capsule is a condensed, well defined layer

closely surrounding the cell, The capsule is considered a
5 by Dr.Sana A.Alshakely
M.Sc. Samar M.M.
 URUK UNIVERSITY
PRACTICAL MICROBIOLOGY LAB
THIRD GRADE
virulence factor because it enhances the ability of bacteria to
cause disease (e.g. prevents phagocytosis).

The detection of capsule could be a useful tool in bacterial typing like

Quellung reaction which is used for typing streptococcus pnemoniae.

6 by Dr.Sana A.Alshakely
M.Sc. Samar M.M.
 URUK UNIVERSITY
PRACTICAL MICROBIOLOGY LAB
THIRD GRADE

7 by Dr.Sana A.Alshakely
M.Sc. Samar M.M.