Clostridium tetani

teaching basics
Dr.T.V.Rao MD

08/23/15

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Clostridia:
general characteristics
Genus Clostridium contains a large
number of gram-positive, spore-forming
species, several of which are able to
produce disease in humans.
Most species are obligate anaerobes,
some will grow under microaerophilic
conditions.
Natural habitat: soil and the intestinal
Tract
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History
Tetanus was well known to ancient
people, who recognized the
relationship between wounds and fatal
muscle spasms. In 1884, Arthur
Nicolaier isolated the strychnine-like
toxin of tetanus from free-living,
anaerobic soil bacteria.
Rosenbach and Kitasato contributed
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Dr.T.V.Rao MD @ Rao's Microbiology

Clostridium tetani
Clostridium tetani is a rodshaped, anaerobic bacterium of the
genus Clostridium.. C. tetani is found
as spores in soil or as parasites in the
gastrointestinal tract of animals. C.
tetani produces a potent biological
toxin, tetanospasmin, and is the causative
agent of tetanus
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Dr.T.V.Rao MD @ Rao's Microbiology

Clostridium tetani
Clostridium tetani is
an anaerobic
pathogenic bacterium
that is primarily found
in soil and animal
intestinal tracts.
Cotton, dust
Plaster of Paris
Catugut
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Microbiology

Morphology
Gram + bacilli
4-8 x 0.5 microns
Drumstick
appearance
Obligatory
anaerobe

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Microbiology

Cultural characteristics'
Grows on blood agar with advancing
edges
In gelatin stab fir tree appearance
In Robertson cooked meat medium
produces turbidity gas
Meat is not digested turns black
First hemolysis followed by hemolysis
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Dr.T.V.Rao MD @ Rao's Microbiology

C. tetani: key characteristics

Large, spore-forming,
motile, obligate anaerobic
bacillus (see below).
Ferments: proteins or
amino acids.
Produces: acetic acid,
fatty acids, NH3, CO2, H2,
and a strong exotoxin.
Tetanospasmin, a
powerful neurotoxin
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Dr.T.V.Rao MD @ Rao's
Microbiology

Resistance of the Clostridium tetani

.Survives boiling upto
3 hours
They can not
survive autoclaving
at 249.8 F (121 C)
for 20 minutes.
The spores are also
relatively resistant
to phenol and other
chemical agents.
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Dr.T.V.Rao MD @ Rao's
Microbiology

Biochemical reactions
Indole +
MR and VP test
negative
H2 S not produce
Nitrites are not
reduced
Gelatin liqification
occurs
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Microbiology

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Tetanus etiology
In necrotic and infected wounds,
anaerobic conditions will permit
germination. Contaminated
puncture wounds can be
particularly dangerous,
especially when a foreign body
is present.
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Mode of Transmission -

Tetanus
Mode of Transmission: Transmission is
primarily by contaminated wounds,
Tissue injury( surgery, burns,deep
puncture wounds, crush wounds, Otitis
media ,dental infection, animal bites,
abortion, and pregnancy
Incubation Period: 8 DAYS ( 3-21 DAYS)

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Type of Tetanus

Traumatic tetanus
Puerperal tetanus
Otogenic tetanus
Idiopathic tetanus
Tetanus
Neonatroum

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Tetanus Toxin oriented disease

C. tetani usually enters a host through a
wound to the skin and then it replicates.
Once an infection is established, C.
tetani produces two
exotoxins, tetanolysin and tetanospasmin..
The genes that produce toxin are encoded
on a plasmid which is present in all
toxigenic strains, and all strains that are
capable of producing toxin produce
identical toxin
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Nature of Toxins
C. tetani produces two exotoxins,
tetanolysin and tetanospasmin.
The function of tetanolysin is not
known with certainty.
Tetanospasmin is a neurotoxin and
causes the clinical manifestations of
tetanus.
Tetanospasmin estimated Human lethal
dose 2.5 ng/kg
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Toxins

Hemolysin ( tetanolysin )
Neurotoxin ( Tetanospasmin )
Non spasmogenic toxin
Tetanolysin is heat labile and oxygen laible
Toxin is responsible for the Tetanus
Heat laible inactivated at 650c
Mol wt Heavy chain 93,000
Light chain 52,000 bound by disulphide bonds

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Tetanospasmin
Mol wt Heavy chain 93,000
Light chain 52,000
bound by disulphide bonds
Human 130 nanograms
Horse and guinea pigs guinea
pigs rabbits effected
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Tetanospasmin
Tetanospasmin is distributed in the
blood and lymphatic system of the
host. The toxin acts at several sites
within the central nervous system,
including peripheral nerve terminals,
the spinal cord, and brain, and within
the sympathetic nervous system.
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Tetanospasmin
The toxin is taken up into within the
nerve axon and transported across
synaptic junctions, until it reaches the
central nervous system, where it is
rapidly fixed to gangliosides at the
presynaptic junctions of inhibitory
motor nerve ending
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Pathogenicty
The spores germinate in
reduced oxygen potential
devitalized tissues,
presence of foreign
bodies
Motor neurons absorb
Spread Intraaxonally to
CNS
Avidly fixed to
gangliosides of gray
matter
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Microbiology

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Mechanism of Action
of Tetanus Toxin

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Tetanospasmin
Resembles strychnine blocks synaptic
inhibition in the spinal cord
At inhibitory terminals that use glycine and
GABA as neurotransmitters
Toxin acts presynaptically unlike
styrchnine post synaptically
Muscle spasms of agonists and
antagonists
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How Toxin acts

The toxin, by blocking the release of
inhibitors, keeps the involved muscles
in a state of contraction and leads to
spastic paralysis , a condition where
opposing flexor and extensor muscles
simultaneously contract. Death is
usually from respiratory failure.
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Clinical manifestations
The clinical manifestations of tetanus
are caused when tetanus toxin blocks
inhibitory impulses, by interfering with
the release of neurotransmitters,
including glycine and gammaaminobutyric acid. This leads to
unopposed muscle contraction and
spasm.
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Tetanus

Involves somatic musculkar system

Injuries
Punctured wound
Surgery
Otitis media
Septic abortion
Cow dung applications
Ear boring circumcision
Incubation 2 days to weeks
Average 6 12 days

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Microbiology

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Opisthotonus

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Microbiology

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Photo Courtesy of U.S. Centers for Disease

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Control and Prevention

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Tetanus symptoms & signs

Characteristic
features are risus

sardonicus (a
rigid
smile), trismus (comm
only known as "lockjaw"), and

opisthotonus
(rigid, arched back).
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Microbiology

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Risus Sardonicus in Tetanus Patient

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Laboratory Diagnosis
of Tetanus

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Laboratory Diagnosis
Clinical diagnosis most important
in management
Microscopy
Culture
Animal Inoculation
Microscopy not reliable
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Culture
Done on Blood agar
One half of plate spreads to other half after 1 2
days of incubation anerobically
Three tubes of cooked meat broth inoculated
1 tube 800c for 15 mt
2 tube 800c for 5 mt
3 tube unheated
All are incubated at 370c upto 4 days
Subcultured
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Clostridium tetani Gram Stain

Round terminal spores give cells a drumstick or tennis

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racket

Showing spores and Grwoth on

Blood Agar plate

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Microbiology

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Toxigenicty tests
In a plate of Blood divided into 2 halves first half
incorporated with 1500 / ml of antitoxin
Hemolysis without antitoxin
Bacteria grown in Robertson cooked meat
medium inoculated into tail of a mice
A 2nd animal injected with tetanus antitoxin 1000
units an hour earlier the test
Spasm of tail of un inoculated mice spread to
limb and other side and the animal dies
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Tetanus bacteria grow in RCM

Medium

Bacteria
grown in
Robertson
cooked meat
medium
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Microbiology

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Treatment
Treatment involves debridement, the antibiotic
metronidazole* active immunization with
tetanus toxoid , and passive immunization with
tetanus immune globulin.
Prevention is through active immunization with
tetanus toxoid**. The toxoid stimulates the body
to make neutralizing antibodies against the
binding component of the tetanus toxin. Once
the antibody binds to the toxin, the toxin can no
longer bind to the receptors on the host cell
membrane
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Prevention
Tetanus carries a 35% mortality rate, making prevention very important!
The best course is childhood immunizations, with consistent booster doses,
and prompt cleaning of wounds with hydrogen peroxide.

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Microbiology

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Prophylaxis

Surgical attention
Antibiotics
Immunization
Antibiotics <4 hours
Erythromycin 5000 mg Bid
Pencillin
Local treatment with bacitracin, and
neomycin

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Passive Immunization
Tetanus antitoxin 1,500 IU s/c IM
Test dose to be given
Human Anti tetanus globulin 250
units

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Passive Immunization
Antitoxin (tetanus immune globulin) should be
administered immediately. This will inactivate toxins in
the blood.
Wounds should be debrided to remove dead tissue or
foreign bodies.
Antibiotics should be given to inhibit growth of C. tetani.
A tetanus toxoid booster immunization should be given
to patients who have not received one within the last 5
years.
If spasms occur, antispasmodic drugs should be used
and respiration maintained by a breathing apparatus if
necessary.
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TETANUS TOXOID
Tetanus toxoid was developed by Descombey in
1924,
Tetanus toxoid immunizations were used
extensively in the armed services during World
War II.

Although the rates of seroconversion are

about equal,the adsorbed toxoid is
preferred because the antitoxin response
reaches higher titers and is longer lasting
than that following the fluid toxoid.
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Tetanus toxoid
Tetanus toxoid
consists of a
formaldehyde-treated
toxin.
There are two types
of toxoid available
adsorbed (aluminum
salt
precipitated)toxoid
and fluid toxoid
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Microbiology

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Active Immunization

1st dose
2nd dose
3rd dose
1st booster
2nd booster
3rd booster

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6th week
10th week
14th week
18th month
6th year
10th year

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Passive Immunization
1. ATS(equine) Ig- 1500 IU/s.c after
sensitivity test
(or)
2. ATS(human) Ig- 250-500 IU, no
anaphylactic shock, very safe and
costly.
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Treatment of Tetanus patients

Hospitalization
Tracheotomy
Human TIG 10,000 IU
Pencillin, Metronidazole
ATS IV
Give active immunization.

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Microbiology

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PREVENTION OF NEONATAL
TETANUS
2 doses of T.T to all pregnant women between
16 to 36 weeks of pregnancy with an interval of
1 to 2 months between the two doses.
The first dose as early as possible & the
second dose a month later preferably 3 weeks
before delivery.
If the pregnant woman is previously
immunized, a booster dose is sufficient.
If the pregnant woman is not immunized, then the new
born should be protected against tetanus by giving
tetanus human immunoglobulin 750 IU with in 6 hours
of birth.
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Triple Antigen vaccine

Triple Antigen vaccine
is a combination of
Diphtheria, Tetanus,
and Pertussis. The
vaccine stimulates the
production of
antibodies to
immunize the body
against the causative
agents of the three
diseases listed above
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Microbiology

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Epidemiology
Endospores are found in most soils and in the
intestinal tract of many animals and humans.
Although exposure to endospores is common,
disease is uncommon except in countries with
poor medical care and vaccination compliance.
It is estimated that there is more than one million
cases a year worldwide, with a mortality rate of
20% to 50%.
Most deaths occur in neonates and originates
from infection of umbilical stumps in mothers
that have no immunity.
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Prevention is Better than

Cure

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Microbiology

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 Program Created and Designed by

Dr.T.V.Rao MD for the benefit of
Universal Education on Infectious
diseases
Email
doctortvrao@gmail.com

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