LOVELY

PROFESSIONAL
UNIVERSITY

TERM PAPER OF
ADVANCED
MICROBIOLOGY

Submitted to: -
Submitted by:-

Ms POOJA TANDAN ISHA SINGH

MICROBIOLOGY Msc
Microbiology
 Roll no: -
12

CLOSTRIDIUM TETANI

CLASSIFICATION :-

Division-Prokaryotes
Kingdom - Bacteria
Phylum - Firmicutes
Class - Clostridia
Order - Clostridiales
Family - Clostridiaceae
Genus - Clostridium
Species - Clostridium tetani

INTRODUCTION:-

Clostridium tetani was discovered in 1884 by a German physician Arthur Nicolaier. He found
that C.tetani causes tetanus when he injecting test animals with garden soil which contain
bacteria. Clostridium tetani is an anaerobic bacterium that is found in soil and animal
intestinal tracts. C. tetani bacteria are single-celled and do not contain any membrane-bound
organelles, such as a nucleus. It is a Gram-positive bacteria as it lacks an outer
lipopolysaccharide membrane and possesses only a thick peptidoglycan cell wall.C.tetani is a
rod-shaped bacteria and have flagella in vegetative form and drumstick in shape in its spore
form. Eleven strains of C.tetani are identified and all of these strains produce an neurotoxin
known as tetanospasmin. This toxin is responsible for Tetanus.[1]

As Clostridium tetani bacteria are anaerobic so they derieve nutrition by the process of
fermentation. Acetic acid, butyric acid, butanol and acetone are byproducts of the bacterial
fermentation. Certain strains of C. tetani produce different enzymes that are used to
externally digest organic material in preparation for fermentation. In the anaerobic condition
C. tetani is primarily found in soil, manure, and intestinal tracts. [2]
Clostridium tetani bacteria have two primary life stages, sporular and vegetative. The latter
stage is extremely anaerobic and bacteria of this state are unable to survive oxygen exposure
where as sporular form is quite hardy and can withstand oxygen and environmental extremes.
Clostridium tetani reproduces by an asexual reproduction process known as binary fission. In
this process a single bacteria cell after reaching maxium growth stage divides into two
smaller identical clone bacteria cells. Due to this process, C. tetani growth is exponential.

Figure: Clostridium tetani in the sporular state[3].

C. tetani usually enters a host through a wound to the skin and then it replicates. On entering
the host C. tetani produces two exotoxins known as tetanolysin and tetanospasmin. Eleven
strains of C. tetani have been identified which only differ in flagellar antigens and their
ability to produce tetanospasmin. The genes that produce toxin are encoded on a plasmid
which is present in all toxigenic strains.Tetanolysin serves no known function to C. tetani.
Tetanospasmin is a neurotoxin and causes tetanus. Tetanus toxin is generated in living
bacteria and is released when the bacteria lyses such as during spore germination or during
vegetative growth. A minimum amount of spore germination and vegetative cell growth are
required for toxin production. Tetanospasmin is a zinc-dependent metalloproteinase which is
similar in structure to botulinum toxin but each toxin have different effects. C. tetani [which
remain connected with a disulfide bridge. Cleavage of the tetanospasmin toxin into A and B
fragments can also be induced artificially with trypsin.If infections of C. tetani left untreated
or ineffectively treated then they may result in the commonly fatal tetanus. Tetanus is a
potentially deadly nervous system disease. Tetanus causes muscular spasms (tightening of the
muscles) that can cause paralysis of the respiratory (breathing) system and lead to death. The
disease is also known as lockjaw because there is a a common symptom of tetanus in which
the jaw muscles become tight and rigid and a person becomes unable to open his mouth.
Sometimes tetanus affects only one part of the body but usually the infection spreads
throughout the body until the entire body becomes paralyzed. The incubation period for
tetanus is from two to fifty days. The incubation period is the time between infection and the
first appearance of symptoms. There are increased chances of death if symptoms occur early.
In neonatal tetanus, symptoms usually appear from 4 to 14 days after birth, averaging about 7
days.

There are mainly three types of tetanus Local tetanus, Cephalic tetanus and generalised
tetanus. Local tetanus is an uncommon form of the disease, in which patients have persistent
contraction of muscles in the same anatomic area where the injury is present. The
contractions may persist for many weeks before gradually subsiding. Local tetanus is
generally milde only about 1% of cases are fatal, but it may lead to the generalized tetanus.
Cephalic tetanus is a rare form of the disease and is occasionally occurring with otitis media
(ear infections) in which C. tetani is present in the flora of the middle ear or following
injuries to the head. There is involvement of the cranial nerves, especially in the facial
area.Generalized tetanus is the most common type of tetanus representing about 80% of
cases.. The first sign is trismus or lockjaw and the facial spasms called risus sardonicus,
followed by stiffness of the neck, difficulty in swallowing, and rigidity of pectoral and calf
muscles. Other symptoms include elevated temperature, sweating, elevated blood pressure,
and episodic rapid heart rate. Spasms may occur frequently and last for several minutes with
the body shaped into a characteristic form called opisthotonos. Spasms continue for 3–4
weeks, and complete recovery may take months.[4]

Figure: case of untreated tetanus [4]

Figure: map of the world telling about the concentration of tetanus cases. Darker area
represents high concentration of tetanus cases. [5]

METHODOLOGY:-

The infection begins when endospores of C. tetani enter an anaerobic wound. Tetanus can
also occur in burn victim, in patients receiving intramuscular injections, person obtaining a
tattoo, in persons with frostbite, dental infections, penetrating eye injuries and umbilical
stump infections. Other risk factors include diabetes, chronic wounds(skin ulcers.gangrene)
parental drug abuse and recent surgery .Unvaccinated mother, home delivery, and unhygienic
cutting of the umbilical cord increase susceptibility to tetanus. History of neonatal tetanus in
a previous child is a risk factor for subsequent neonatal tetanus. Potentially infectious
substances applied to the umbilical stump (e.g., animal dung, mud, clarified butter) are risk
factors for neonates. Immunity from tetanus decreases with advancing age. Since the
bacterium is an obligate anaerobe, an anaerobic environment is needed for the endospores to
germinate and the vegetative bacteria to grow. Vegetative bacteria eventually produce
tetanospasmin which is responsible for symptoms of tetanus. The tetanus toxin initially binds
to peripheral nerve terminals. It is transported within the axon and across synaptic junctions
until it reaches the central nervous system. There it becomes rapidly fixed to gangliosides at
the presynaptic inhibitory motor nerve endings and is taken up into the axon by endocytosis.
The effect of the toxin is to block the release of inhibitory neurotransmitters (glycine and
gamma-amino butyric acid) across the synaptic cleft which is required to check the nervous
impulse. If nervous impulses cannot be checked by normal inhibitory mechanisms it produces
the generalized muscular spasms characteristic of tetanus. The toxin appears to act by
selective cleavage of a protein component of synaptic vesicles synaptobrevin II and this
prevents the release of neurotransmitters by the cells. Major symptoms of tetanus spasms and
tightening of the jaw muscle known as lockjaw, stiffness and spasms of various muscle
groups like neck muscles, chest muscles, abdominal muscles, back muscles.Painful,powerful
muscle contraction,irritability,fever.Additional symptoms that may be associated with this
disease are excessive sweating,swallowing,hand or foot swamps, uncontrolled urination[6]

DIAGNOSIS: -

The symptoms of tetanus are quite distinctive and a doctor can diagnose the disease simply
by observing the patient. Painless spasm of the muscles of the jaw, difficulty in opening the
mouth, contraction of the muscles on the forehead and mouth are the characteristic
symptoms. Knowing whether or not the patient has had a tetanus vaccination also helps a
doctor make their diagnosis. Other signs of being generally irritable, muscle spasms, and
poor ability to take in liquids (poor sucking response), Irregular heartbeat, changes in blood
pressure

Tetanus can be tested by spatula test. This simple test involves touching the oropharynx with
a spatula or tongue blade. This test typically elicits a gag reflex and the patient tries to expel
the spatula (which indicates a negative test result). If tetanus is present patients develop a
reflex spasm of the masseters and bite the spatula (which indicates a positive test result). [6]
TREATMENT:-

When a tetanus infection becomes established, treatment usually focuses on controlling

muscle spasms stopping toxin production, and neutralizing the effects of the toxin. Treatment
includes administration of tetanus immune globulin (TIG) which comprises antibodies which
inhibit tetanus toxin (also known as tetanus antitoxins) by binding to and removing unbound
tetanus toxin from the body. Binding of the toxin to the nerve endings is an irreversible event,
and TIG is ineffective at removing bound toxin. Recovery of affected nerves requires the
sprouting of a new axon terminal. Large doses of antibiotic drugs (such as metronidazole or
intramuscular penicillin G) are also given once tetanus infection is suspected, to halt toxin
production. The wound must be cleaned. Dead and infected tissue should be removed by
surgical debridement. Metronidazole treatment decreases the number of bacteria but has no
effect on the bacterial toxin. Penicillin was once used to treat tetanus, but this treatment is no
longer used, owing to a theoretical risk of increased spasms. However, its use is
recommended if metronidazole is not available. Passive immunization with human anti-
tetanospasmin immunoglobulin or tetanus immune globulin is crucial. If specific anti-
tetanospasmin immunoglobulin is not available, then normal human immunoglobulin may be
given instead. All tetanus victims should be vaccinated against the disease or offered a
booster shot.

An infant suffering from neonatal tetanus.

Treatment of tetanus is based on whether it is mild or sever tetanus. Mild cases of tetanus can
be treated with: Tetanus immune globulin IV or IM ,metronidazole IV for 10 days ,diazepam
tetanus vaccination .
 Lock-jaw in a patient suffering from tetanus.

Human tetanus immunoglobulin injected intrathecally, tracheotomy and mechanical

ventilation for 3 to 4 weeks, magnesium as an intravenous (IV) infusion to prevent muscle
spasm, diazepam (Valium) as a continuous IV infusion, the autonomic effects of tetanus can
be difficult to manage like alternating hyper- and hypotension, hyperpyrexia/ hypothermia
and may require IV labetalol, magnesium, clonidine, or nifedipine. Drugs such as diazepam
or other muscle relaxants can be given to control the muscle spasms. In extreme cases it may
be necessary to paralyze the patient with curare-like drugs and use a mechanical ventilator. In
order to survive a tetanus infection, the maintenance of an airway and proper nutrition are
required. An intake of 3500-4000 Calories, and at least 150g of protein per day, is often given
in liquid form through a tube directly into the stomach, or through a drip into a vein. This
high-caloric diet maintenance is required because of the increased metabolic strain brought
on by the increased muscle activity. [7]

PREVENTION:

There are two important ways to prevent tetanus: getting vaccinated against tetanus along
with other routine immunizations, or, after an injury that could cause tetanus, receiving a shot
(post-exposure tetanus prophylaxis).For kids, tetanus immunization is part of the DTaP
(diphtheria, tetanus, and a cellular pertussis) vaccinations. They typically receive a series of
four doses of DTaP vaccine before 2 years of age, followed by a booster dose at 4 to 6 years
of age. After that, a booster (Tdap) is recommended at 11 to 12 years of age, and then a
tetanus and diphtheria booster every 10 years through adulthood. Post-exposure tetanus
prophylaxis also involves getting tetanus shots, but after an injury occurs. Shots given will
depend on the number of years since the patient's last booster, the total number of tetanus
vaccinations the patient has received, and the nature of the wound. The doctor may
recommend a tetanus booster Td, DTaP, or Tdap, depending on the patient's age and previous
immunizations and an injection of tetanus immune globulin (TIG) to neutralize any toxin
released by the bacteria. Neonatal tetanus can be prevented by making sure that all pregnant
women have had their tetanus immunizations, by delivering babies in sanitary conditions, and
by proper umbilical cord care. If you are pregnant, discuss your immunization record with
your obstetrician well before your due date. Any skin wound — especially a deep puncture or
a wound that may be contaminated with feces, soil, or saliva — should be cleaned and
dressed right away. Although it's important to clean all wounds, remember that cleaning is
not a substitute for immunization [8].

RESULTS AND DISCUSSION:

Tetanus remains a severe disease occurring primarily among persons who are unvaccinated or
inadequately vaccinated. Adults aged greater than or equal to 60 years continue to be at
highest risk for tetanus and for severe disease. However, the overall incidence of tetanus has
decreased slightly since the late 1980s and early 1990s. Older adults are at highest risk for
tetanus because of the low prevalence of immunity to tetanus. Tetanus remains a clinical
diagnosis because confirmatory laboratory tests are not available for routine use. Isolation of
the organism from wounds is neither sensitive nor specific: anaerobic cultures of tissues or
aspirates usually are not positive, and the organism might be grown from wounds in the
absence of clinical signs and symptoms of disease. Tetanus is preventable through both
routine vaccination and appropriate wound management. Vaccination with a primary series of
three doses of TT-containing vaccine and booster doses of Td every 10 years are highly
effective in preventing tetanus.
CONCLUSION:-

Tetanus (commonly called lockjaw) is a disease manifested by uncontrolled muscle spasms.

Tetanus is caused by the toxin (poison) produced by the bacteria, Clostridium tetani. This
disease is frequently fatal, especially to the very old or very young. Tetanus is preventable by
immunization or through administration of tetanus immune globulin.It occurs predominantly
in developing countries among newborn infants, children, and young adults, especially in
unimmunized or inadequately immunized adults over 50 years of age. The symptoms are
often jaw or neck stiffness, difficulty swallowing, and irritability. There may be pain or
tingling at the site of inoculation (where the tetanus bacteria entered), followed by spasticity
of nearby muscles. The diagnosis is made clinically by the medical history and physical
findings. Immune globulin, given intramuscularly, is the immediate treatment of
unimmunized individuals exposed to material likely to contain the tetanus bacteria. Treatment
includes bed rest and quiet conditions.Antimicrobial drugs, such as penicillin, are used to
eradicate the bacteria. Tetanus is completely preventable by active immunization.
REFERENCES:-

1. http://en.wikipedia.org/wiki/Clostridium_tetani
2. http://bioweb.uwlax.edu/bio203/s2008/unrein_bren/nutrition.
htm

3. http://bioweb.uwlax.edu/bio203/s2008/unrein_bren/adaptati
on.htm

4. http://www.textbookofbacteriology.net/clostridia.html

5. http://kidshealth.org/parent/infections/bacterial_viral/tetanu
s.html

6. http://www.umm.edu/ency/article/000615.html

7. http://www.cdc.gov/vaccines/vpd-vac/tetanus/default.htm

8. http://en.wikipedia.org/wiki/Tetanus