Lab 9

Proteus
General character:

• Rod shape.
• gram-negative.
• motile.
• non-capsulated .
• Possessing flagella.
• Non-lactose fermenting.
-Proteus species are found in the intestinal tract
of human.

- Species of medical importance:

• P. mirabilis
• P. vulgaris
 P. mirabilis
causes 90% of all Proteus infections in humans.
Clinical features:
• Urinary tract infection
• Septicemia
• Abdominal and wound infection
• Secondary invader of ulcer, burn and chronic
discharging ear.
 P. vulgaris
• Important nosocomial pathogen.
• Isolated in wound infection and urinary tract
infection.
 Laboratory diagnosis

Specimen

• Urine.
• Pus.
• blood.
• ear discharge
 Gram stain
• Gram-negative rods
Culture characteristic
 On blood agar

• Proteus mirabilis does not form distinctive colonies

on Blood Agar, instead the bacteria swarm across the
surface of the agar.

• P. mirabilis produces a very distinct fishy odour.

• Discontinuous swarming produces concentric circles
around the point of inoculation.
• Continuous swarming produces a uniform film.
On nutrient agar
 On MacConkey agar
• Non-lactose fermenters that may exhibit swarming

colorless colonies with slight swarming

On EMB
Biochemical test
• Indole test
is used to differentiate
P.mirabilis (-ve) from
P. vulgares(+ve)
 Urease test
• positive urease (which is the fundamental test to
differentiate Proteus from Salmonella).

• Most strains produce a powerful urease enzyme, which

rapidly hydrolyzes urea to ammonia and carbon monoxide.
 Gelatinase test
• Positive result.
• Liquefaction of gelatin by gelatinase enzyme.
• oxidase-negative

• Catalase-positive.
 Lab 8

Pseudomonas
• Medically important Pseudomonas:
P. aeruginosa.
• present in small numbers in the normal intestine
flora and on the skin.
• Commonly present in moist environments in
hospitals.
• It is primarily a nosocomial pathogen.
 General character
• Gram-negative rods.
• Motile by polar flagella.
• aerobics.
• Grow well at 42⁰C.
• Non-lactose fermenting
 Pigment production
• can produce pigments, such as:
• Pyocyanine (blue-green)
• Pyoverdin (fluorescent yellow- greenish pigment)
• Pyorubrin (red)
• Pyomelanin (brown)
 Laboratory diagnosis
Specimen:
• Wound discharge
• sputum
• Blood
• Urine
• CSF
 Microscopy
-Gram stain
Gram negative rod
undistinguished from
enerobacteriaceae.
On Culture media
 Culture character

• Form smooth and round colonies.

• Fluorescent greenish colour.
• production of fruity odor (grape-like) .
• Inability to ferment lactose.
 On nutrient agar
• Colonies are smooth, large, translucent
• Greenish blue diffusible pigment
 On blood agar
• Grayish colonies
• Many are haemolytic
(beta hemolysis).

This picture shows Pseudomonas aeruginosa

(on the right) and Shigella dysenteriae (on the
left) in blood agar.
 On MacConkey agar
• Non-lactose fermenting (colourless colonies)
 On EMB
• Non lactose fermenting
 Pigment production on Cetrimide agar
• is a type of agar used for the selective isolation of
the gram-negative bacterium, Pseudomonas aeruginosa.

• As the name suggests, it contains cetrimide, which is the

selective agent against alternate microbial flora.
• Cetrimide also enhances the production
of Pseudomonas pigments such as pyocyanin and fluorescein,
which show a characteristic blue-green and yellow-green
colour, respectively.
Pseudomonas aeruginosa fluorescence under UV illumination
Oxidase test
• Oxidase positive

Catalase test
• Catalase positive