Corynebacterium

diphtheriae
 Corynebacterium diphtheriae
• Morphology
– Non-sporing, non-capsulated, and non-motile
– Gram +ve rod with a tendency to clubbing at one or both
ends
– Exist extensively in nature. In human, they are localized
on skin and upper respiratory tract.
– Attach to each-other in pairs, resembling the letters "V"
or "L", or in palisade.
– Have metachromatic granules composed of
polyphosphate
Corynebacterium diphtheriae. A photomicrograph of
Corynebacterium diphtheriae, showing pleomorphism (especially
club forms), metachromatic granules, and palisades arrangement.
Corynebacterium diphtheriae
Corynebacterium diphtheriae
• Culture
– Aerobic and facultatively anaerobic
– Löffler's serum medium is commonly used
– Tellurite-blood-agar is a selective medium
containing potassium tellurite (0.04%)
 Diphtheria
• Epidemiology
– Diphtheria is worldwide in distribution where there is
overcrowding.
– The sources of infection are active infections and
carriers.
– Transmission occurs from person to person by
respiratory droplets or skin contact.
– Cutaneous diphtheria also occurs
 Pathogenesis
• Virulence factor
– Exotoxin: C. diphtheriae produces systemic
manifestations from production of exotoxin, also
called as diphtheria toxin.
• Synthesis: only strain of C. diphtheriae that have
been infected with β-phage carrying the tox gene
produces exotoxin.
• The toxin inhibits protein synthesis in a host cell.
 Diphtheria
– The toxin affects the body on two levels.
• Locally, it produces an inflammatory reaction (low-grade
fever may occur as a result), sore throat, nausea,
vomiting, enlarged cervical lymph nodes, and severe
swelling in the neck (Bull neck appearance).
• One life-threatening complication is the
pseudomembrane, that develops in the pharynx from the
solidification of fluid (exudate) expressed during
inflammation. The pseudomembrane is so leathery and
sticky that attempts to pull it away result in bleeding, and
if it forms in the airways, it can cause airway blockage.
Diagnosing diphtheria. The clinical appearance in diphtheria infection
includes gross inflammation of the pharynx and tonsils marked by grayish
patches (a pseudomembrane) and swelling over the entire area.
 Diphtheria
– Toxemia occurs when the toxin is absorbed from the
throat and carried by the blood to certain target organs,
primarily the heart and nerves.
– The action of the toxin on the heart causes
myocarditis.
– Peripheral nerve : Cranial and spinal nerve involvement
can cause muscle weakness and paralysis.
– Although toxic effects are usually reversible, patients
with inadequate treatment often die from respiratory
obstruction, or heart damage.
 Diphtheria
droplet、 Local infection by
C. diphtheria contamination oropharynx bacteria and toxinGray-white
trachea pseudomembrane
2-7 day incubation
period

toxemia Produces
mucous
edema and
these may
Cardiac muscle Peripheral block
myocarditis nerve respiratory
•Paralysis of tract
palate and
dysphagia
 Immunization
• Active immunization
– After infection, there occurs lifelong immunity
– Can be immunized with toxoid (DPT)
– Infants are passively protected with maternal
antibodies
• Passive immunization
– Given antitoxin subcutaneously
 Schick test

• This test is done to check whether the person is having

immunity against diphtheria or not.
• A dose of diluted diphtheria toxin (0.1 ml) is injected
intradermally in one forearm and a control dose of heat-treated
toxin is injected into other forearm. The test is read at 24 and
48 hours, and again in 6 days.
• Positive reaction: It indicates absence of neutralizing antitoxin.
Redness and swelling will occur on arm at the site of injection
and disappear after 6 to 7 days.; no reaction on control arm.
– Not protected.
• Negative reaction: No reaction at either site of injection:
antibody present.
– Protected.
 Laboratory Diagnosis
• The initial diagnosis of diphtheria is entirely clinical
• Specimens: can be taken from throat or skin
• Smears: morphology, metachromatic granules
• Culture: the Löffler’s media, tellurite blood agar
 Treatment and Prevention
• Prevention
– Active immunization: by using DPT
– Passive immunization: by using antitoxin
– Chemoprophylaxis: Penicilli to eliminate the organism
• Treatment
– Diphtheria antitoxin
– Antibiotics: penicillin
n­
Bacillus anthracis
 Bacillus anthracis
• Bacillus anthracis is the causative organism of
anthrax, a disease primarily confined to animals,
and humans gets infected secondarily.
• A lot of “firsts”:
– It is the first pathogenic bacteria to be seen under
the microscope.
– The first communicable disease to be transferred
experimentally by inoculation of infected blood
– The first bacteria used for preparation of
attenuated vaccine.
Morphology and identification
• Gram + ve,
• Large and rectangular cell.
• Spore forming bacilli.
• Non-motile
• Capsules are formed in the tissue but are usually
lacking in culture: polypeptidal in structure
• The spores are oval and central in position and its
diameter is same as the width of the bacillus.
– Spore are formed in soil when the organism is shed
by the animal or grown on culture media. Spores are
highly resistant to killing.
Bacillus anthracis
Morphology and identification
• The bacilli are aerobic and grow well in ordinary media
producing large granular greyish white colonies with a
frosted glass appearance in nutrient agar.
• Under the low power microscope, the edge of the
colonies are composed of long, interlacing chains of
bacilli resembling locks of matted hair. This is called
medusa head appearance.
• Non-hemolytic on blood agar.
Morphology and identification

Medusa head appearance

 Virulence factors
• Capsule helps organism to resist phagocytosis but
antibodies are not protective.
• Exotoxin is produced only when the bacteria is
growing inside the tissues. The toxin consists of
three protein components (maximum toxicity
occurs when all three components are present).
– Protective antigen
– Lethal factor
– Edema factor
 Virulence factors
– Protective antigen (PA) reacts with host tissue
receptors where it is activated so that it binds LF and
EF to allow entry of LF and EF into the host cells.
Injection of PA into a host will result in the production
of antibody against PA that provides short term
immunity.
– Lethal factor (LF Both PA and LF are required for
lethal activity.
– Edema factor (EF) Both PA and EF are required for
edema to occur.
)­
 Anthrax
• Anthrax, the disease caused by B. anthracis, is essentially
a disease of animals who acquire the organism by
ingestion or inhalation of spores. The spores are
extremely resistant to adverse chemicals and physical
environments. They may remain a source of infection in soil
for 20-30 years.
• Goat and sheep are the most common sources of anthrax
followed by cattle and horses.
• Human infection is acquired during dealing with
contaminated products of animals e.g., goat hair, wool etc.
So, it is an example of zoonotic disease.
 Anthrax
– The type of anthrax depends upon the mode of
transmission:
• Pulmonary (Woolsorter‘s disease):
– Spores are inhaled and germinate in the lungs where they
multiply and spread to cause a fatal septicemia or
meningitis. This is the most serious form of the disease.
• Intestinal anthrax:
– Results from ingestion of spores.
– Rare type
– bloody vomiting, diarrhea and abdominal distension occurs.
• Cutaneous anthrax:
– This is the most common form of anthrax . Here organisms
come in through skin abrasions.
– Forms black necrotic membrane like lesion known as eschar.
 Anthrax
• Eschar: Appears on
exposed parts of body
 Anthrax
• Eschar: means dried and
thick crust
– Spore germinates at
site of entry
– Eschar looks black and
spreading
– Painless
– May heal
spontaneously
• May cause septicemia
and mortality in 20% of
patients.
 Diagnostic Laboratory Tests
• Specimens
– Fluid or pus from local lesion, blood, sputum
• Stained smears
– Chains of large Gram-positive rods are often seen,
also can be identified by immunofluorescence staining
technique.
• Culture
– gray colonies with typical microscopic morphology
can be seen on blood agar plate. (frosted
appearance)
 Treatment
• Penicillin or tetracycline

• Prevention:
– A short-term PA vaccine is available for industrial
workers and others at high risk.
 Bacillus cereus
• Bacillus cereus is a large, Gram-positive, rod-
shaped, endospore forming, facultative aerobic
bacterium
• It is distributed widely in nature and is commonly found
in the soil.
• As a soil bacterium, B. cereus can spread easily to
many types of foods such as plants, eggs, meat, and
dairy products, and is known for causing 2-5 % of
food-borne intoxications due to its secretion of
enterotoxin.
• Can cause diarrhea and vomiting (emesis).
 Bacillus cereus
• The diarrheal type is associated with a wide range
of foods, has an 8 to 16 hour incubation time, and is
associated with diarrhea and gastrointestinal pain.
– Enterotoxin can be inactivated after heating at 56 °C for
5 minutes.
• The 'emetic' form is commonly caused by rice
cooked for a time and temperature insufficient to kill
any spores present, then improperly refrigerated.
– It can produce a toxin, cereulide, which is not inactivated
by later reheating. This form leads to nausea and
vomiting one to five hours after consumption.
– Emetic toxin can withstand 121 °C for 90 minutes
 Listeria
• It is facultatively anaerobic and motile bacteria.
• It can grow and reproduce inside the host's cells.
• It is one of the most virulent food-borne pathogens.
• Listeria monocytogenes is the most common species
• Causes listeriosis.
– Listeriosis primarily causes infections of the central nervous
system (meningitis, meningoencephalitis, brain abscess) and
bacteremia in those who are immunocompromised, pregnant
women, and those at the extremes of age (newborns and the
elderly), as well as gastroenteritis in healthy persons who
have ingested a large amount of the organism.
• Treatment includes prolonged administration of
antibiotics, primarily ampicillin and gentamicin.
 Actinomyces
• Anaerobic bacteria
• Gram positive rods
and have branching
filaments
• Not stained by acid
fast stain
• Culture:
– Thioglycate broth (A.
israelli): fluffy balls
appearance
– Solid media like blood agar:
spidery colonies
 Actinomyces
• Actinomyces israelii:
– Commonly found in human gingival crevices.
– Pathogenesis:
• Invasive growth in tissue with compromised oxygen supply,
anaerobic growth.
• An affected human often recently had dental work, poor oral
hygiene, periodontal disease or trauma (broken jaw) causing
local tissue damage to the oral mucosa, all of which
predispose the person to developing actinomycosis.
• Disease:
• Actinomycosis
– Cause draining abscess with sulphur granules in
exudate that can be use for culture.
– It occur in the form of lumpy jaw, thoracic and pelvic
abscess, brain abscess.
 Nocardia
• Nocardia asteroides:
– Aerobic.
– Gram positive branching
rods.
– It grows readily on ordinary
media, forming dry,
granular, wrinkled colonies.
– It produces pigment ranging
from yellow to red.
– Partially acid fast.
– Reservoir: soil, dust.
– Transmission: airborne or
traumatic implantation.
 Nocardia asteroides

• Pathogenesis:
– Immunosuppression

• Disease:
– Nocardiosis ( cavitatory bronchopulmonary
nocardiosis)

– Cutaneous/subcutaneous nocardiosis ( starts with

traumatic implantation, causes subcutaneous abscess
with granules