OBJECTIVES

Practical No.3
➢ To study microscopic appearance of staphylococci

➢To identify cultural characteristics of staphylococci.

Identification of ➢To differentiate between staphylococci species.

Staphylococcus Species ➢To perform antibiotic sensitivity tests for
staphylococci.

Dr. Banan Atwah

Morphological characteristics Cultural Characteristics: S.aureus

Gram Reaction: On blood agar:-

• G +ve cocci
• Golden to cream or
occasionally dirty
Cell arrangement: white colonies
• Arranged in bunches
• 1 – 2 mm in diameter
(clusters)
• Slightly raised

Cultural Characteristics:
Cultural Characteristics: S.epidermidis
S.aureus (Contd.)

On blood agar:- On blood agar:-

• some strains are - • white

hemolytic

• golden colour colonies • non-haemolytic

colonies.

•Novobiocin
sensitive
Cultural Characteristics: S.saprophyticus
Biochemical Reactions
On blood agar:-

• non-hemolytic
• Catalase test

• bright white • Coagulase test

• creamy colonies • DNase test
• Novobiocin
resistant
• Mannitol fermentation test

Catalase Test Coagulase Test

➢ Positive test:
Appearance of gas
bubbles

➢ Negative test:
NO bubble
➢ Positive test: ➢ Negative test:
production
Macroscopic clumping in No clumping in
10 seconds or less in plasma drop.
plasma drop.

Coagulase Test DNase Test

1. Positive test: ➢ Positive DNase

Clotting of Test:
2
plasma
Presence of clear
halos around
1 colonies indicates
2. Negative test: presence of DNase.
No Clotting of
plasma
Mannitol Fermentation Test Differentiation of Species
Characteristics S.aureus S.epidermidis S.saprophyticus
➢ On Mannitol Salt
Agar (MSA): Colour of colonies Yellow to white White to pale
white gray
Acid production due Haemolysis Some cause No haemolysis No haemolysis
-haemolysis
to fermentation of
Coagulase production +ve -ve -ve
mannitol turns the
phenol red indicator DNase production +ve -ve -ve
yellow.
Mannitol +ve -ve -ve
fermentataion
+ve for S.aureus
Novobiocin sensitivity Sensitive Sensitive Resistant

Antibiotic Susceptibility Test:

(Kirby Bauer Disc Diffusion Test)

Clear zones around

antibiotic discs
THANKS
indicate sensitivity to
respective antibiotic.