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Study of Proteus Spp.

The document outlines a study on Proteus spp., highlighting their association with infections such as UTIs and sepsis. It details the methodology for isolating and identifying these bacteria through staining and biochemical tests, including Gram staining and catalase tests. The provisional diagnoses suggest the presence of either Proteus mirabilis or Proteus vulgaris based on the test results.

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subarnachowdhury64
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6 views4 pages

Study of Proteus Spp.

The document outlines a study on Proteus spp., highlighting their association with infections such as UTIs and sepsis. It details the methodology for isolating and identifying these bacteria through staining and biochemical tests, including Gram staining and catalase tests. The provisional diagnoses suggest the presence of either Proteus mirabilis or Proteus vulgaris based on the test results.

Uploaded by

subarnachowdhury64
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Download as PDF, TXT or read online on Scribd
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Date: ……………………………

TOPIC: Study of Proteus spp.

Competency Number: MI 1.1

Competency addressed: Describe the different causative agents of infectious diseases, methods used
in their detection and discuss the role of microbes in health and disease.

Name of activity: Isolation of bacteria and Identification of common pathogenic bacteria: by staining
& their biochemical tests

Introduction:

Proteus spp. are occasionally encountered in human lesions. They are associated with:

1. UTI, especially following catheterisation or cystoscopy


2. Sepsis: Abdominal, wound and middle ear infections. The organisms are generally low
grade pathogens and often are secondary invader of ulcers, pressure sores, burns and
damaged tissues.
3. Septicaemia

Requirements:

I. Equipment: Compound microscope, Bunsen burner and Bacteriological loop


II. Consumables:
a) Glassware: Clean grease-free slides
b) Reagents:
1. Gram’s stain: 0.85% saline or normal saline, crystal violet (basic dye), Gram’s iodine
(mordant), acetone (decolourising agent) and safranine (counter stain).
2. Catalase test: 3% H2O2 (v/v)
c) Miscellaneous: Marker pen

Technique:

Culture plate 1

Macroscopical examination
a) Identification of culture medium: MacConkey’s agar

b) Colony characteristics:

 Size: Large (1-3 mm X 0.5 mm)


 Shape: Circular
 Surface: Smooth
 Elevation: Flat
 Margins: Entire regular edge
 Translucency: Opaque
 Colour: Pale colonies due to non-lactose fermentation
 Odour: Fishy smell
 Consistency: Moist
 Emulsification: Easily emulsifiable
Tests: We performed the following tests:

a) Gram’s stain:

Pink coloured rod-


shaped bacteria
seen

Fig: Gram stained smear showing pink coloured rod shaped bacteria

Observation: Pink coloured rod-shaped pleomorphic bacteria seen


Interpretation: Gram-negative bacilli seen

b) Catalase test:

Fig: Catalase test

Observation: Bubbles was formed immediately


Inference: Positive reaction

c) Motility test by hanging drop:


Bacteria
moving from
one place to
another place

Fig: Hanging drop motility test showing motile bacteria

Observation: Bacteria are moving from one place to another.


Inference: Motile bacilli are seen.
d) Biochemical reactions:

Indole: Negative
Glucose: Fermented and production of acid as well as gas (AG)
Lactose: Not Fermented
Sucrose: Not fermented
Triple Sugar Iron (TSI): K/A with H2S gas
Citrate: Negative (Citrate is not utilised)
Urease: Positive (Urease enzyme is produced)
Phenylalanine deaminase or Phenyl pyruvic acid (PPA) test: Immediate appearance of
an intense green color after adding 4 or 5 drops of Ferric chloride reagent

e) Provisional Diagnosis: It may be Proteus mirabilis

Culture plate 2

Macroscopical examination
a) Identification of culture medium: MacConkey’s agar
b) Colony characteristics:
 Size: Large (1-3 mm X 0.5 mm)
 Shape: Circular
 Surface: Smooth
 Elevation: Flat
 Margins: Entire regular margin
 Translucency: Opaque
 Colour: Pale colonies due to non-lactose fermentation
 Odour: Fishy smell
 Consistency: Moist
 Emulsification: Easily emulsifiable

Tests: We performed the following tests:


a) Gram’s stain:

Pink coloured rod-


shaped bacteria
seen

Fig: Gram stained smear showing pink coloured rod shaped bacteria

Observation: Pink coloured rod-shaped pleomorphic bacteria seen


Interpretation: Gram-negative bacilli seen
b) Catalase test:

Fig: Catalase test

Observation: Bubbles was formed immediately


Inference: Positive reaction

c) Motility test by hanging drop:

Bacteria
moving from
one place to
another place

Fig: Hanging drop motility test showing motile bacteria

Observation: Bacteria are moving from one place to another.


Inference: Motile bacilli are seen.

d) Biochemical reactions:

Indole: Positive
Glucose: Fermented and production of acid as well as gas (AG)
Lactose: Not Fermented
Sucrose: Not Fermented
Triple Sugar Iron (TSI): K/A with H2S gas
Citrate: Negative (Citrate is not utilised)
Urease: Positive (Urease enzyme is produced)
Phenylalanine deaminase or Phenyl pyruvic acid (PPA) test: Immediate appearance of an
intense green color after adding 4 or 5 drops of Ferric chloride reagent

e) Provisional Diagnosis: It may be Proteus vulgaris

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