Corynebacterium

Diphtheria

Prof.
Zainab A. Aldhaher
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 Corynebacterium
• Gram + Non Acid fast, Non motile,
• Irregularly stained with granules,
• Club shaped swelling at one or both ends
so the name
• Important Pathogen
Corynebacterium diphtheria,
Diptheros meaning leather,

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• It’s a localized infectious disease,
which usually attacks the throat
and nose mucous membrane

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• C. diphtheriae is an aerobic gram-
positive bacillus.
–Pleomorphic, club-end
–Non-spore-forming
–Non-acid-fast
–Non-motile

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• Definition
• •Diphtheria is an acute infectious disease that
typically strikes the upper respiratory tract
including the throat. It is caused by infection
with the bacteria Corynebacterium
diphtheria. It’s characterized by sore throat
and mild fever at first. As the disease
progresses, a membranous substance forms
in the throat that makes it difficult to breathe
and swallow.
Causes,,,,
• Corynebacterium diphtheriae causes
diphtheria. Usually the bacteria multiply on
or near the surface of the mucous
membranes of the throat, where they
cause inflammation.
• The inflammation may spread to the voice
box (larynx) and may make your throat
swell, narrowing your airway. Disease-
causing strains of C. diphtheriae release a
damaging substance (toxin), which can also
involve the heart, brain and nerves.
• Pathogenesis
• C. diphtheria infects the respiratory tract
by droplet inhalation or infects the skin by
direct contact.
• Most clinical infections are contracted from
carriers rather than symptomatic patients.
• C. diphtheria carriers usually harbor the
organism in their throat or nose.
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• The organism doesn’t penetrate deeply
into mucosal tissue and bacteremia
doesn’t occur
• The organism starts to multiply at the site
of entry and produces small amount of
exotoxin to kill any phagocytic cell and
then multiplies freely causing inflammation
of the throat with local necrosis
• At the same time the organism secretes
exotoxin to the blood

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 Diphtheria Exotoxin
• It is a heat labile polypeptide composed of
two fragments : A (active) and B (binding)
• Virulence is due to an extoxin
• It inhibits protein synthesis – kills cells
• Causes necrosis (death) of cells in upper
respiratory tract – Pseudomembrane
formation
• Toxin can be distributed systemically to
cause life – threatening damage to organs
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• Signs and symptoms
• •A sore throat
• •Painful swallowing
• •Swollen glands (enlarged lymph nodes) in
your neck

• Signs and symptoms usually begin two to five

days after a person becomes infected.
• •A thick, gray membrane covering your
throat and tonsils
• •Difficulty breathing or rapid breathing
• •Nasal discharge
• •Fever and chills
• Cutaneous diphtheria occurs in tropical
countries, the lesion is characterized by a
non healing ulcer covered by necrotic
pseudomembrane

Dr.T.V.Rao MD 17
 Etiology
• There are three biotypes —
gravis, Intermedius, and
mitis. The most severe
clinical type of this disease
is associated with the gravis
biotype, but any strain may
produce toxin. 18
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 Staining methods
• Grams method
• Albert's stain
• Neissers stain
• Ponders stain
• On staining seen as
Pairs, Appear as v and L letters, resembling
Chinese letter pattern or also called
cuneiform arrangement.
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 Cultural characters
• Need enrichment Media
• Contain
• Blood, Serum or Egg 37 c ph 7.4
• Aerobic/Facultative anaerobic.
• Commonly used medium
• Loeffler serum slope,
• Tellurite Blood agar,

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Gram +ve Bacilli and Colonies

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 Culturing
• Selective & differential medium
• Corynebacterium are resistant to
tellurite
– Reduced to tellurium
• Forms deposit in colonies
– Colonies appear dark
• Biotypes
– gravis, Intermedius, mitis

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 Growing on Culture Plates
• Loffler serum slope
Grows rapidly in
6 -8 hours,
Small white
opaque disks
Turns to yellow
Tellurite blood agar
Modified Mac
Leod
Hoyles medium.
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 Commonly used medium

• Tellurite blood agar

Contains tellurite
0.04 tellurite
Inhibits other
bacteria
• Produce
Grey/Black
colonies.

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Thick Membrane

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Pseudo membrane

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 Laboratory Diagnosis
• Specific treatment is more
important than Laboratory
Diagnosis.
1.Isolation of Diphtheria bacilli.
2.Testing for toxigenicity
3.Tissue culture test
4.Polymerase chain reaction
5.Elisa test
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 Collection of Specimens
• Throat swabs
• Smear examinations
Gram s staining,
Albert's, Ponders
Immunoflorescent methods
Cultures on Loeffers serum slope
Tellurite Blood agar,
Blood agar.
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 Isolation of C.diptheria
• Serum slope – Growth in 6 – 8 hours,
• Stain with Neissers stain Albert's
stain
• Bacilli have metachromatic granules,
• Tellurite Blood agar takes two days
for manifestation of colonies,

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 Toxigenicity Tests
In Vitro Elek test
In Vivo Animal
inoculation
rabbit skin test-
necrosis
guinea pig
challenge test-
lethal
low [Fe 2+] induces toxin
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Schick Test ( Out dated )
–Schick test: It is an intradermal test,
the test is carried out by injecting
intradermally into the skin of
forearm 0.2 ml of diphtheria toxin,
while into the opposite arm is
injected as a control, the same
amount of toxin which has been
inactivated by heat.
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 Interpretation
• Negative reaction: If a person had immunity to diphtheria,
no reaction will be observed on either arm.

• Positive reaction: An area of in duration 10-15 mm in

diameter generally appears within 24-36 hours reaching
its maximum development by 4-7 days, the control arm
shows no change. The person is susceptible to diphtheria.

• False positive reaction: A red flush develops in both arms,

the reaction fades very quickly, and disappears by 4th day.
This is an allergic type of reaction found in certain
individuals
• Combined reaction: the control arm shows pseudo
positive reaction and the test arm is true +ve reaction,
susceptible and need vaccination 35
 Schick Test
• Injection of toxin I
D
• Produces
redness/erythemati
c in 2-4 days
• No reaction –
Protective
immunity present.

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 Active Immunization.
• Toxoid – Toxin treated with Formaldehyde
• Absorbed Toxoid
• Given by Intramuscular route
• Given in DTP –Triple Vaccine
• Primary Immunization
• Three Doses of DPT at least 4 weeks apart.
• Non vaccinated
• Three doses of Toxoid four weeks apart
• One dose after One Year.

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 Prevention
Vaccination: Immunisation with diphtheria toxoid,
combined with tetanus and pertussis toxoid (DTP
vaccine), should be given to all children at two,
three and four months of age. Booster doses are
given between the ages of 3 and 5 .

The child is given a further booster vaccine

before leaving school and is then considered to
be protected for a further 10 years (16 – 18
years).

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 Passive Immunization
• Given in Acute infections
• Give Subcutaneously
• 500 – 1000 Units of Antitoxin
• Given as Horse Serum
• Combined in Acute Infections ( Both
Active Immunization with Toxoid and
Antitoxin.
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 Treatment
• Commonly used antibiotics,
• Penicillin parentally,
• Oral Erythromycin

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