CORYNEBACTERIUM

Maya Savira, MD, M.KES

Microbiology Department
Riau University
 CORYNEBACTERIUM

The genus name was established to accommodate the

diphtheria bacillus

Coryne come koryne (Greek = pallisade )

 MORPHOLOGY
Pleomorphic form: Rod coccoid, rudimentary
branching may occur but no definite mycelia.
Gram (+)
Arranged in angular V and L shape like Chinese
Letter.
Non spore forming but with Babes Ernt granule
Non Acid fast
Motility may be (+).
Species of the medical important :
1. C. diphtheriae ( Diphthery ).
2. C. ulcerans ( exudative pharyngitis and diphtheria like disease, nasopharynx-
carrier ).
3. C. Pseudotuberculosis ( Granulomatous lymphadenitis ).

Those three species produced diphtheria toxin.

4. C. haemolyticum ( pharyngitis and skin ulcer, occasionally mimic diphthery )

5. C. xerosis ( endocarditis, bacteremia, pneumonia, surgical wound inf. )
6. C. pseudodiphthericum ( endocarditis, UTI )
Those species other than C. diphtheriae have
reported to cause tissue and blood stream in-
infection, including :
1. Bacteremia.
2. Endocarditis.
3. Osteomyelitis.
4. Upper Resp. Infection.
5. Wound infection.
Diphtheroid Bacteria.
This term has been used in medical bacteriology
for : Gram (+) Rods that resemble and may
be confused with. C. diphtheriae and
are presumably a species of genus Co-
rynebacterium. With metachromatic
granules. It is usually the commensal
inhabitants of the skin
in Morphologic Based Taxonomic, Corynebac-
terium could be found in human, animal, plant
and saprophytic form.
by Chemotaxonomic based is used, the Coryne
bacterium contain :
1. Meso-diaminopimelic acid.
2. Arabinose.
3. Galactose.
4. short chain mycolic acid
( 22 38 Carbon atom )
Corynebacterium was found only in human
and animal ( based on the chemotaxonomic )

Metabolism : Facultative anaerob

It is striking similarities in the cell wall compo-

sition with Mycobacterium and Nocardia
Corynebacterium diphtheriae
Pathogenesis
Transmission mostly by droplets, usual portal
of entry is the upper resp. tract. Attached in
mucosal cell layer with its pili, multiplied in the
mucous membrane layer, produced and ela-
borated their toxin ( exotoxin )
On the upper resp. tract mucous membrane the
exotoxin causes necrosis of the adjacent tissue.
Triggering inflammatory response result in for-
mation of dihtheritic pseudomembrane.

Toxin synthesis in the local lesion is absorbed

and carried by the blood to the all parts of hu-
man body, but the toxic effect involved prima-
rily the heart, and peripheral nerve.
Diphtheritic Pseudomembrane, composed of :
1. Bacteria
2. Necrotic epithelium
3. Phagocytes
4. Fibrin.
the membrane usually appears first on the
tonsil or posterior pharynx, then extend either
upward into soft and hard palate, nasopharynx
and down ward to larynx and trachea.
Laryngeal diphtheria is particularly dangerous.
Cutaneous diphtheria is common in the tropics.
During outbreak of diphtheria cases it is recom-
mended to routinely culture the bacteria from
the skin of patients and contact persons.

Skin lesion due to C. diphtheriae is usually

secondarily infected abrasion or insect bite
which also contain Streptococcus or Staphylo-
coccus aureus or both.
The significance of C. diphtheriae in skin lesion
may be its role as a reservoir for these patho
gens during epidemic.
It is also the opportunity of genetic exchange
with coryneform bacteria in the skin.

Diphtheritic lesions also occur in the nares an-

terior, inner nose, mouth, eye, middle ear, and
in rare cases in vagian.
Laboratory Diagnostic Procedure
Specimen
In both patient and contact person, the speci-
men is swabs taken from the nasopharynx as
well as the throat, especially under the pseudo-
membrane, 20 % of positive cultures can be missed
if only one site is cultured.
Nasipharyngeal swab shoud be obtained with
a flexible alginate swab that reaches deep into
the posterior nares.
In Suspected cases of diphtheria, swab of the
throat are taken with cotton swab which is firm
ly apply to any area with a membrane or inflam-
mation.
For asymptomatic patients, the tonsillar fossae,
posterior pharynx and retrouvular should be
sampled as well as nasopharynx.
In case of wound infection the lesions should be
cleansed with sterile normal saline, and crusted
material should be moved.
 SPECIMEN TRANSPORT

The specimen should be directly send to the

lab. If there is a delay, specimen shoud be
streak to Loffler medium and could be sent
im a day or more.
Microscopic Examination

Special staining for Diphtheric bacilli is needed.

Albert staining method is currently used for this
purpose. An experience laboratory technician
can easily detected suspected bacteria with
greenish rod in palisade arrangement and con-
tain Babes Erns granules in one side or both like
a clubbed end or a halter.
CULTURE
Specimens are inoculated onto a Loeffler or Pai
slant, a cystein- tellurite agar plate, and a 5 %
sheep red blood agar plate.
Incuation for overnight at 35* C
Methylen blue staining could be done and see
the bacteria microscopically and reported to
the clinician if any suspected diphtheric bacilli
detected.
Any suspected colonies then subcultured to
loeffler and tellurite agar and followed by bio-
chemical test.
Another medium can also be used instead of
loeffler or tellurite, the agar is Tinsdale medium
Colony identification
There are 3 types of colony could be found on
culture medium : Gravis, Intermedius and mitis
Those types of colony reflects the severity of
disease produced by each type.

Gravis is very severe clinical outcome.

Intermedius less severe and not fatal.
mitis gave mild symptoms even without symp-
toms at all.
Biochemical test

C. diphtheriae is catalase (+), Nitrate (+), Urea

(-), Glucose and maltose (+) with gas. Trehalo
se and sucrose (-), mannitol and xylose (-)
Esculin hydrolysis (-), gelatin hydrolysis (-)
Toxinogenicity Test
All isolates of C. diphtheriae are tested for toxin
production. There are methods available :

1. The in vitro immunodiffusion test.

2. Guinea pig subcutaneous lethality test.
3. In vitro modified Elek immunodiffuion test