Pyogenic bacteria

By
M.SC Marwa.A.Ahmed

• Staphylococcus ssp
• Streptococcus ssp
• Neisseria ssp
 Staphylococcus aureus
• Gram-positive
• Common inhabitant of the skin and mucous membrane.
• Lack of spores and flagella.
• May have capsules.
 Specimen collection

Specimen collection
• Pus from pyogenic lesions.
• Blood from septicemia.
• Cerebrospinal fluid from meningitis
• Sputum from respiratory infection
• suspected food, vomit, or faces from food poisoning.
• Mid-stream urine in urinary tract infection.
• An anterior nasal swab from suspected carriers.
 Cultural
1. Blood agar
• Grows in large, round, colonies.
• The optimum temperature of 37 C
• Facultative anaerobe
• Isolation is best done in blood agar.
• Produce a characteristic golden yellow carotenoid pigment.
• On blood, agar colonies are usually surrounded by a zone of clear
hemolysis.
 2.Mantoil salt agar(MSA)

• Differentiation between staph. aureus and staph .epidermitis.

• Positive = acid end products turn the phenol red pH indicator from
red to yellow.
• negative = phenol red remains red.

Mantoil salt agar Staph.aureus Staph.epidermitis Staph.saprophyticus

Result positive Negative positive

3. Nutrient agre

• golden yellow opaque colonies

with smooth
 Microscope examination

• Gram-positive
• Spherical cells arranged in an
irregular crystal.
• like grapes
 Biochemical test
1. Coagulase test
• Detects the extracellular enzyme “free
coagulase” or staphylocoagulase.
• Causes a clot to form when bacterial cells
are incubated with plasma.
• Procedure
• Inoculate rabbit plasma with the organism
and incubate at 35-37
• observe at 30 minutes for the presence of
a clot Continue for up to 24 hours, if
needed
Coagulase test Staph.aureus Staph.epidermitis Staph.saprophyticus

Result positive Negative Negative

2. Catalase test

• Principle: tests for the enzyme

catalase
• 2 H2O2 2 H2O + O2

• Procedure
• Smear a colony of the organism
to a slide.
• Add drop H2O2( hydrogen
peroxide) onto smear
Catalase test Staph.aureus Staph.epidermitis Staph.saprophyticus

Result positive Negative Negative

 2. Gelatinase test

• Nutrient gelatin is a differential medium that

tests the ability of an organism to produce an
exoenzyme (extracellular molecules), called
gelatinase that hydrolyzes gelatin to
polypeptide and releases amino acid.
• The gelatinase test can be used to differentiate
between Staphylococcus aureus and
Staphylococcus epidermidis.
gelatinase test Staph.aureus Staph.epidermitis

Result positive Negative

 2. DNase test
• Use DNA agar to Detect Dnase enzyme
extracellular molecules Positive = clear
zone around growth after adding 1N HCl
(no DNA remaining in the agar)
hydrolase Hcl to carbon.
• negative = cloudy around growth after
adding 1N HCl (DNA remains in the agar
forming a precipitate)
Dnase test Staph.aureus Staph.epidermitis Staph.saprophyticus

Result positive Negative Negative