volutin

Corynebacterium diphtheriae

Corynebacterium diphtheriae is a pathogenic bacterium that causes diphtheria. It is also known as the Klebs-Löffler bacillus, because it was discovered in 1884 by German bacteriologists Edwin Klebs (1834 – 1912) and Friedrich Löffler (1852 – 1915).

Morphology and toxin production

C. diphtheriae is a aerobic organism Gram positive organism, characterized by non-encapsulated, non-sporulated, immobile, straight or curved rods with a length of 1 to 8 µm and width of 0.3 to 0.8 µm, which form ramified aggregations in culture (looking like "Chinese characters")and sometimes which have clubbed ends[Gr.Coryne,a club] . The bacterium may contain polymetaphosphate aggregates called Volutin granules. It is pathogenic only in humans. C. diphtheriae produce diphtheria toxin, a proteic exotoxin, with a molecular weight of 62 kilodaltons which ADP-ribosylates host EF-2, resulting in the inhibition of protein synthesis and thus is responsible for the signs of diphtheria. The inactivation of this toxin with an antitoxic serum (antitoxin) is the basis of the antidiphtheric therapeutic vaccination. However, not all strains are toxigenic; the ability to produce the exotoxin is conferred on the bacterium when it is infected by a bacteriophage (a mechanism termed "lysogenic activation"). A non-toxigenic strain can thus become toxigenic by the infection of such a bacteriophage.

Classification

Four subspecies are recognized: C. diphtheriae mitischodis, C. diphtheriae intermedius, C. diphtheriae gravis, and C. diphtheriae belfanti. The four subspecies differ slightly in their colonial morphology and biochemical properties such as the ability to metabolize certain nutrients, but all may be toxigenic (and therefore cause diphtheria) or non-toxigenic.

Diagnosis

In order to accurately identify C. diphtheriae, a Gram stain is performed to show gram-positive, highly pleomorphic organisms with no particular arrangement (classically resembling Chinese characters). Then, culture the organism on an enrichment medium, namely Löffler's serum, to allow it to overgrow any other organisms present in the specimen. After that, use a selective plate known as tellurite agar which allows all Corynebacteria (including C. diphtheriae) to reduce tellurite to metallic tellurium producing brown colonies and, only in the case of C. diphtheriae, a black halo around the colonies allowing for easy differentation of the organism.

A low concentration of iron is required in the medium for toxin production. At high iron concentrations, iron molecules bind to an aporepressor on the beta bacteriophage which carries the genes for the Tox gene, converting it to a repressor which shuts down toxin production. This is most appreciated when performing Elek's test for toxogenecity, in order to know if the organism is able to produce the diphtheria toxin or not.