MORPHOLOGY AND CULTURE CHARACTERISTICS OF CLOSTRIDIUM PERFRINGENS (C. WELCHII OR BACILLUS WELCHII)

 MORPHOLOGY OF CLOSTRIDIUM PERFRINGENS (CLOSTRIDIUM WELCHII)

ShapeClostridium perfringens is a large, rod shape (bacillus) bacterium with straight, parallel sides and rounded or truncated ends.

Size – The size of Clostridium perfringens is about 4–6 µm × 1 µm (micrometer).

Arrangement Of CellsClostridium perfringens is arranged singly, in short chains or in small bundles. They are pleomorphic, filamentous & involution forms are commonly present.



MotilityClostridium perfringens is a Non-motile bacterium.

FlagellaClostridium perfringens is a Non-flagellated bacterium

Spores – The Central or Sub-terminal spores are present in C. perfringens but are rarely seen in artificial cultures or in the microscopic examination of lesions. The spore formation takes place on a selective medium used for the cultivations of this bacterium.

CapsuleC. perfringens is a capsulated bacterium. The capsule can easily be demonstrated using India ink preparation, which appear as a clear halo in a dark background.

Gram Staining Reaction – Clostridium perfringens is a Gram +ve (Positive) bacterium.

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CULTURE REQUIREMENTS OF CLOSTRIDIUM PERFRINGENS (CLOSTRIDIUM WELCHII)

Special requirementsClostridium perfringens or C. welchii have complex nutritional requirements and readily grow in a media containing Blood or Egg, commonly Blood Agar medium & Robertson’s Cooked meat broth medium is used for the cultivation of Clostridium perfringens in Laboratory.

 Optimum temperatureClostridium perfringens ranges from 20 –50°C but usually grown at 37°C in laboratories. Also, 45°C is the optimum temperature for some strains of the Clostridium perfringens.

⇒ Optimum pH – 5.5 – 8.0

⇒ Oxygen requirementsC. perfringens (C. welchii) is an anaerobic bacterium i.e. can grow in the absence of oxygen and it is also a microaerophilic organism i.e. can grow in the low oxygen environment.

⇒ There are various culture media used for the cultivation of C. perfringens (C. welchii) in the laboratory and most commonly the Blood Agar medium and CMB medium is used, the other media are as follows –

  • Egg yolk Agar medium
  • Columbia Horse Blood Agar medium
  • MacConkey Agar medium
  • Litmus milk
  • Tryptose Sulfite Cycloserine (TSC) Agar (Selective medium).
  • Tryptone Sulfite Neomycin (TSN) Agar medium a.k.a. Marshal’s medium (Selective medium).
  • The liquid medium (Nutrient Broth medium, TSB medium, etc.).

 The Tryptose sulfite Cycloserine Agar (TSC) medium which is the Selective medium for Clostridium perfringens can be prepared by incorporating Cycloserine alone or along with the 4-Methylumbelliferyl-phosphate disodium salt in Perfringens Agar base or in TSC Agar base medium for the growth of Clostridium perfringens.

 Similarly, the Tryptone Sulfite Neomycin Agar (TSN) medium which is also a selective medium for Clostridium perfringens can be prepared by incorporating Neomycin and Polymixin B Sulfate in the TSN Agar Base medium for the Growth of Clostridium perfringens.

The Litmus milk for the Litmus milk test which is commonly used for the C. perfringens can easily be prepared in the laboratory by suspending Skim milk powder (100gm), Litmus powder (0.5 gm) & Sodium sulfite (0.5gm) in 1000 ml of distilled water.

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CULTURE CHARACTERISTICS OF CLOSTRIDIUM PERFRINGENS (CLOSTRIDIUM WELCHII)



Cultural CharacteristicsBlood Agar Medium (BAM)Tryptose Sulfite Cycloserine (TSC) Agar mediumTryptone Sulfite Neomycin (TSN) Agar medium
ShapeCircular or RoundCircular or RoundCircular or Round
Size3-5 mm2-3 mm2-3 mm
ElevationNo elevation - Flat Colonies----------
SurfaceFlat & FilamentousFlatSmooth
ColorWhite or light yellowBlackBlack
StructureTranslucent –OpaqueTranslucent –OpaqueOpaque
HemolysisTarget Hemolysis----------

In Roberson’s Cooked meat broth medium, C. perfringens grows well and the meat is turned pink or Black as per the type of strain. The Saccharolytic strains of C. perfringens turn the meat pink with sour odor and the Acidic reaction of the medium whereas the Proteolytic species of Clostridium perfringens turn the meat black with the foul smell and Acidic pH of the medium.

In litmus milk test, the fermentation of lactose (sugar present in milk) leads to the formation of acid in the medium, which is visible by the change in color of medium from blue to red and the clotted milk, due to the formation of acid which coagulates the casein (protein present in milk), usually marked by stormy fermentation (the clotted milk disrupted due to the gas produced during fermentation and the pressure of gas pushes it towards the mouth of test tube due to which the casein sticks to the wall of the tubes).

In liquid culture media like Trypticase soy broth or Nutrient broth, the growth of the bacterium occurs as a turbidity in the broth medium, but the growth is scanty in these ordinary media, which is further analyzed for the morphology (under the microscope), gram reaction, biochemical tests, and Clostridium perfringens specific tests.

THAT’S ALL ABOUT THE MORPHOLOGY AND CULTURE CHARACTERISTICS OF CLOSTRIDIUM PERFRINGENS (C. WELCHII OR BACILLUS WELCHII)



Further Reading:

Wikipedia

Microbewiki

Foodborne illness

2 thoughts on “MORPHOLOGY AND CULTURE CHARACTERISTICS OF CLOSTRIDIUM PERFRINGENS (C. WELCHII OR BACILLUS WELCHII)

  1. We are using fluid thioglycholate broth for cl .perfringens to grow under anaerobic conditions.

    After growth if we do plating on Tsc agar with anaerobic conditions,black colour isolated colonies are not getting.

    Can you please suggest.

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