3rd Stage

Lab. 17
2016/2017
By: Mohammed Hmood
Non lactose fermenter bacteria
Proteus & Pseudomonas

Proteus species:
 Gram -ve rods
 Non lactose fermenter
 Non capsulated
 Actively motile by peritrichous flagella resulting in swarming on solid media unless the
swarming is inhibited by chemicals.
 Produce urease resulting in hydrolysis of urea.
 Produce H2S from sodium thiosulfate.
 Produce phenylalanine deaminase.

Diseases
1- Urinary tract infection→ causing alkaline urine.
2- Wound infection.
3- Others (nosocomial infections)

Lab Diagnosis
• Specimen: urine, blood, wound swabs, sputum.
• Cultivation:
1- Mac Agar……….. Non-lactose fermenter
2- Blood agar……. swarming phenomenon

• Biochemical reaction:
1- Urease test
2- TSI

Swarming on Blood agar Lactose non-fermenter on Mac agar

1
 Pseudomonas species
 Gram-ve rods
 Motile with polar flagella
 Obligate aerobe
 Oxidase-positive
 Catalase positive
 Not ferment carbohydrates
 Resistant to multiple drugs
Pseudomonas aeruginosa
 Chemoheterotroph
 Adaptable
 Found in soil, marshes,
 Coastal marine habitats, on plants and animals.

Identification of P. aeruginosa is usually based on:

• Colonial morphology,
• B-hemolysis,
• Oxidase positivity,
• Presence of characteristic pigments
• Sweet odor,
• Growth at 42 oC.
Pigments
1. Pyocyanin- nonfluorescent bluish pigment;
2. Pyoverdin- fluorescent greenish pigment;
3. Pyorubin, reddish pigments
4. Pyomelanin black pigments
Clinical Diseases
• Infections of wounds and burns (blue-green pus), Pulmonary infection, Eye infections:
corneal ulcer, Nosocomial infections, Ear infections…Otitis externa , UTI and Sepsis
Laboratory Diagnosis
• Specimen: skin lesions, pus, urine, blood, spinal fluid, sputum.
• Culture: 1- Blood agar plate
2- MacConkey agar

• Biochemical reaction: Oxidase test, TSI

• Subtyping methods, including phage typing and molecular typing, are available for
epidemiologic purposes.
Oxidase Test
o In a positive test, bacterial cytochrome oxidase oxidizes the colorless reduced
substrate tetramethyl-p-phenylenediamine dihydrochloride (TPDD) forming a dark
purple.
o Streak a small portion of bacterial colony to filter paper soaked with a 1% solution of
TPDD.

2
 o If the streak mark turns purple in 10 sec or less, the spot oxidase test is interpreted as
positive.
o Bacterial colonies should be 18 to 24 hr old.
(TSI) Triple Sugar Iron agar:
• Inoculated organism produces acid butt & alkaline slant
• Pseudomonas------alkaline slant, alkaline butt & no H2S
• Proteus -------- alkaline slant, acid butt & H2S production (black colour)
• Note: alkaline ----- red
acid ---- yellow

Pseudomonas aeruginosa on Nutrient agar with production of pigments

Pseudomonas------Alkaline slant, Alkaline butt & No H2S

Proteus -------- Alkaline slant, Acid butt & H2S
production
Note: Alkaline ----- red
Acid ---- yellow

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 Differentiation between Proteus & Pseudomonas

Test Pseudomonas Proteus

1- biochemical test
Urease -ve +ve
Oxidase +ve -ve

2- Pigment +ve
{ pyocyanin and -ve
pyoverdin}

3- Odor +ve fruity odor +ve fishy odor

(green apple-like).

4- Antibiotics Resistance to most Sensitive to most

resistance antibiotics antibiotics