Corynebacterium Diphtheriae

Morpology

Gram stain: Gram positive, club-shaped bacilli

with Chinese letter appearance.

methylene blue: The presence of polyphosphate

inclusions called metachromatic (or volutin)
granules give the bacilli beaded appearance better
seen when stained with
Corynebacterium diphtheriae ,Methylene blue stain
 Culture

•C. diphtheriae is facultative anaerobe and can

grow on blood agar and Loeffler's serum media.
The selective differential media containing
tellurite. A blood agar plate is also inoculated for
the detection of beta-haemolytic streptococcus.

•Following initial isolation, C. diphtheriae may be

identified as mitis, intermedius, or gravis biotype
by differences in both colony morphology and
biochemical tests.
 Loeffler’s serum
Growth on
Loeffler’s serum
Shows best
morphology
 Pathogenesis of Diphtheria
-Mode of transmission: droplets, or direct contact
-IP : 2 – 5 days
Diphtheria is described as "an upper respiratory
tract illness characterized by
-Local pathology: An adherent pseudomembrane
is formed in the upper respiratory tract (the tonsil,
pharynx, and/or nose) involves necrotic injured
epithelial cells, fibrin network resulting from blood
plasma leak and WBCs & RBCs interlaced with
rapidly-growing C. diphtheriae cells. It may spread
locally causing suffocation.
 Systemic pathology (due to Diphtheria toxin)
The diphtheria bacilli do not invade tissues. They produce the
toxin that is absorbed and disseminated through blood to the
susceptible tissues of the body (heart muscle, nerves).

-Two factors influence the ability of C. diphtheriae to produce

the toxin:
(1) the presence of a lysogenic prophage in the bacterial
(2) low extracellular concentrations of iron.

-The gene for toxin production occurs on the prophage, but a

bacterial repressor protein controls the expression of this
gene. The repressor is activated by iron.
•The toxin produced by the three biotypes of C.
diphtheriae (gravis, intermedius and mitis) is
antigenically the same. The differences in virulence
can be explained by the amount of toxin produced
(being highest in gravis).
 Clinical manifestations

•The disease is acute, and has an insidious onset. It can

involve almost any mucous membrane but Pharyngeal
and tonsillar diphtheria is the most common form. A
grayish-white membrane is formed and may extend to
cover most of the soft palate & may result in respiratory
obstruction.
•The patient appears quite toxic with cervical
lymphadenitis and low grade fever.
•The most frequent complications of diphtheria are
myocarditis and neuritis. Death may occur from heart
failure, airway obstruction, or diaphragmatic paralysis.
bullneck diphtheria
Diphtheria
 Laboratory diagnosis
The diagnosis of diphtheria is primarily clinical,
and treatment should be started if diphtheria
was clinically suggested.Rapid microbiological
confirmation of a clinical diagnosis is crucial:

(1)Because specific treatment is effective only if

administered during the early stages
(2)For epidemiological control purposes
 Laboratory diagnosis ( continue)
• Specimens: Swabs from pseudomembrane.
• Direct detection:
Gram stain: club-shaped bacilli with Chinese
letter appearance..
methylene blue: The metachromatic (or volutin)
granules give the bacilli beaded appearance.
• Cultivation and identification: C. diphtheriae is
facultative anaerobe and can grow on blood agar
and Loeffler's serum media. The selective
differential media containing tellurite. identified as
mitis, belfanti, intermedius, or gravis biotype by differences
in both colony morphology and biochemical tests.
 Toxigenicity testing of C. diphtheriae strains

1. Elek test
2. Tissue culture cytotoxicity tests.
3. Detection of diphtheria toxin gene: by PCR.
4. Detection of diphtheria toxin from clinical
isolates by ELISA.

For diagnosis of carriers the same procedures are

applied except the direct Gram stain.
Elek’s Test
 Treatment
• Diphtheria antitoxin: Diphtheria antitoxin must be
given immediately if diphtheria is strongly
suspected on clinical grounds.. It is given
intramuscularly or intravenously, taking precautions
to avoid hypersensitivity reactions (anaphylactic
shock or serum sickness).
• Antibiotics:Erythromycin or penicillin G
• Respiratory support and airway maintenance may
be needed.
 Prevention and control
1. Active immunization: by Diphtheria toxoid
Diphtheria toxoid is available either combined with tetanus
as pediatric DT or adult Td, or with both tetanus toxoid and
acellular pertussis vaccine as DTaP. Vaccine is given by
intramuscular injection. The usual schedule is a primary
series of 4 doses at 2, 4, 6, and 18 months of age. Booster
dose is recommended every 10 years.

2. Preventive measures: For close contacts, especially

household contacts, a booster of diphtheria toxoid should be
given. Contacts should also receive antibiotics (one injection
of benzathine penicillin or erythromycin for 7 to 10 days).
BACILLUS ANTHRACIS
 Morphology
Bacillus anthracis is very large, Gram-positive rod
arranged in chains. They are facultative anaerobe
nonmotile, encapsulated.
•Capsule formation is usually evident in smears from
infected tissues (in vivo). The capsular material is
detected in the McFadyean reaction,
•The bacterium forms oval central non pulging spores
Spores are formed in vitro & are stained with a special
acid-fast (spore) stain. B. anthracis spores are found
in soil habitats around the world.
 Pathogenesis of anthrax
Mode of transmission: Humans acquire anthrax as a result
of contact with infected animals or animal products. There is
no person-to-person transmission of anthrax
Virulence factors;
-Polypeptide capsule
-Anthrax toxin
The oedema factor (EF) produces oedema.
The lethal factor (LF).
The protective antigen (PA)
 Clinical manifestations

1. Cutaneous anthrax (malignant pustule = ‫الجمرة‬

‫ )الخبيثة‬from handling infected material. Spores
from the soil or a contaminated animal or carcass
inoculate a cut or abrasion, usually on an exposed
area. Approximately 20% of untreated cases of
cutaneous anthrax progress to fatal septicaemia.

2. Intestinal anthrax; from eating infected meat.

3. Pulmonary anthrax; from inhaling spore-laden

dust.
 Bacillus Food Poisoning
Bacillus cereus causes two types of food poisoning:
•emetic form: It has an incubation period of 1 to 6
hours and is characterized by nausea, vomiting and
abdominal cramps. It resembles Staphylococcus
aureus food poisoning This form is caused by a
preformed heat-stable enterotoxin (emetic toxin).
•Diarrhoeal form: It has an incubation period of 8 to
16 hours and manifests primarily by abdominal
cramps and diarrhoea. This type resembles food
poisoning caused by Clostridium perfringens. This
form is mediated by a heat-labile enterotoxin which
activates intestinal adenylate cyclase and causes
intestinal fluid secretion.
 Anthrax and Biological Warfare
• Bioterrorism is specifically defined as the use of biological
agents to inflict disease and/or death on humans, animals or
plants.

• The biological agents thought to be the most likely weapons

of bioterrorists include B. anthracis (anthrax), Francisella
tularensis (tularaemia), Yersinia pestis (plague), variola virus
(smallpox), agents of viral haemorrhagic fevers, and
botulism toxin

• The inhalation of anthrax spores can lead to infection and

disease. The possibility of creating aerosols containing
anthrax spores has made B. anthracis a chosen weapon of
bioterrorism.