BANGABANDHU SHEIKH MUJIBUR RAHMAN

SCIENCE AND TECHNOLOGY UNIVERSITY,

GOPALGANJ-8100.

Assignment On
Clostridium botulinum infection.
Course Tittle: Fisheries Microbiology
Course Code:
Submitted by: Submitted To:
Name:Rifath Ahamed Shirin Akter
ID:18FMB065 Assistant Professor,
SESSION:2018-2019 Department of Fisheries & Marine
Department of Fisheries & Bioscience ,
Marine Bioscience, BSMRSTU.
BSMRSTU.

Date of Submission: 30 July 2021.

INTRODUCTION:
Clostridium botulinum is an anaerobic spore-forming gram-
positive bacillus, which is ubiquitous in the
environment. C.botulinum produces eight botulinal
neurotoxins  (BoNT) , some of which can cause disease in
humans. BoNTs are some of the most potent neurotoxins
known to man.  The best known are its neurotoxins,
subdivided in types A-G, that cause the flaccid muscular
paralysis seen in botulism. They are also the main paralytic
agent in botox. C. botulinum is an anaerobic spore-former,
which produces oval, sub terminal endospores and is
commonly found in soil.

Taxonomy history:
C. botulinum was first recognized and isolated in 1895
by Emile van Ermengem from home-cured ham implicated in
a botulism outbreak The isolate was originally named Bacillus
botulinus, after the Latin word for sausage, botulus. ("Sausage
poisoning" was a common problem in 18th- and 19th-century
Germany, and was most likely caused by botulism.)
However, isolates from subsequent outbreaks were always
found to be anaerobic spore formers, so Ida A. Bengtson 
proposed that the organism be placed into the
genus Clostridium, as the genus Bacillus was restricted
to aerobic spore-forming rods.
Since 1959, all species producing the botulinum neurotoxins
(types A–G) have been designated C. botulinum. Substantial
phenotypic and genotypic evidence exists to
demonstrate heterogeneity within the species. This has led to
the reclassification of C. botulinum type G strains as a new
species, C. argentinense.
Group I C. botulinum strains that do not produce a botulin
toxin are referred to as C. sporogenes.
The complete genome of C. botulinum has been sequenced
at Wellcome Trust Sanger Institute in 2007

Characters of C.botulinum:
 A gram-positive (at least in early stage of growth)
 Anaerobic
 Spore forming.
 Toxin forming.
 Heat sensitive.
 Prefers low acid environment.
 Cause Botulism disease.
 Defined as a biothreat level A organism.
Microbiology:
 Toxin produced by the bacterium Clostridium botulinum.
 Anaerobic, gram positive, rod- shaped bacteria.
 Bacteria are 0.5 to 2.0 mcm in width and 1.6 to 22.0
mcm in length.
 Create spores that can remain dormant for 30 years or
more.
 Spores extremely resistant to environmental stressors,
such as heat and UV light.
 Produces botulinum toxin.

Fig: C. botulinum
Toxins:
 liberated during growth.
 Seven types of toxins (A-G).
 Antigenic (light and heavy chain).
 Environmental survival. (Inactivated by heat 100ºC for
20min ).
 Most potential biological warfare agents.
 Lethal dose= 1-2 g .

Neurotoxin types:
Neurotoxin types production is the unifying feature of the
species C. botulinum. Seven types of toxins have been
identified and allocated a letter (A-G). Most strains produce
one type of neurotoxin but strains producing multiple toxins
have been described. The toxin type has been designated as
the type B toxin was found in excess to the type F. Similarly,
strains producing Ab and Af toxins have been reported. There
is evidence that the neurotoxin genes have been the subject of
horizontal gene transfer, possibly from a viral source. This
theory is supported by the presence of integration sites
flanking the toxin in some strains of C. botulinum. However,
these integrations sites are degraded indicating that the C.
botulinum acquired the toxin genes quite far into the
evolutionary past.
Botulinum toxin:
 Botulinum toxin is synthesized as a single polypeptide
chain ( low potency).
 The toxin is nicked by a bacterial protease (or by gastric
proteases) to produce two chains :
 a light chain (the A fragment)
 a heavy chain (the B fragment)
 The A fragment of the nicked toxin, becomes the most
potent toxin found in nature.
 Absorbed in GIT, blood, peripheral neuromuscular
synapse.
 Presynaptic block Acetylcholine release (by proteolysis
of SNARE proteins in neuron which is important in
Ach release) muscle paralysis.
 SNARE proteins are synaptobrevin, SNAP-25 and
syntaxin.

Pathology:
Foodborne botulism:
Signs and symptoms of foodborne botulism typically begin
between 18 and 36 hours after the toxin gets into your body,
but can range from a few hours to several days, depending on
the amount of toxin ingested:
 Double vision.
 Blurred vision.
 Dropping eyelids.
 Nausea, vomiting, and abdominal cramps.
 Slurred speech.
  Trouble breathing.
 Difficulty in swallowing.
 Dry mouth.
 Muscle weakness.
 Constipation.
 Reduced or absent deep tendon reactions, such as in the
knee.

Wound botulism:
Most people who develop wound botulism inject drugs
several times a day, so it's difficult to determine how long it
takes for signs and symptoms to develop after the toxin enters
the body. Most common in people who inject black tar heroin,
wound botulism signs and symptoms include:
 Difficulty swallowing or speaking.
 Facial weakness on both sides of the face.
 Blurred or double vision.
 Dropping eyelids.
 Trouble breathing.
 Paralysis.

Infant botulism:
If infant botulism is related to food, such as honey, problems
generally begin within 18 to 36 hours after the toxin enters the
baby's body. Signs and symptoms include:
 Constipation (often the first sign)
 Floppy movements due to muscle weakness and trouble
controlling the head
 Weak cry
  Irritability
 Drooling
 Dropping eyelids
 Tiredness
 Difficulty sucking or feeding
 Paralysis

Adult intestinal toxemia:

A very rare form of botulism that occurs by the same route as
infant botulism but is among adults. Occurs rarely and
sporadically. Signs and symptoms include:
 Abdominal pain
 Blurred vision
 Diarrhea
 Dysarthria
 Imbalance
 Weakness in arms and hand area

Beneficial effects of botulinum toxin:

Purified botulinum toxin is diluted by a physician for
treatment:
 ongenital pelvic tilt
 Spasmodic dysphasia (the inability of the muscles of the
larynx)
 Achalasia (esophageal stricture)
 Strabismus (crossed eyes)
 Paralysis of the facial muscles
 Failure of the cervix
 Blinking frequently
  Anti-cancer drug delivery

Use and detection:

C.botulinum is used to prepare the medicaments Botox,
Dysport, Xeomin, and Neurobloc used to selectively paralyze
muscles to temporarily relieve muscle function. It has other
"off-label" medical purposes, such as treating severe facial
pain, such as that caused by trigeminal neuralgia.
Botulinum toxin produced by C. botulinum is often believed
to be a potential bioweapon as it is so potent that it takes
about 75 nanograms to kill a person (LD50 of
1 ng/kg, assuming an average person weighs ~75 kg);
1 kilogram of it would be enough to kill the entire human
population. For comparative purposes, a quarter of a typical
grain of sand's weight (350 ng) of botulinum toxin would
constitute a lethal dose for humans.
A "mouse protection" or "mouse bioassay" test determines the
type of C.botulinum toxin present using monoclonal
antibodies. An enzyme-linked immunosorbent assay (ELISA)
with digoxigenin -labeled antibodies can also be used to
detect the toxin, and quantitative PCR can detect the toxin
genes in the organism.
Diagnosis:
 Foodborne botulism: serum analysis for toxins by
bioassay in mice should be done, as the demonstration of
the toxins is diagnostic.
 Wound botulism: isolation of C. botulinum from the
wound site should be attempted, as growth of the bacteria
is diagnostic.
  Adult enteric and infant botulism: isolation and growth
of C. botulinum from stool samples is diagnostic. Infant
botulism is a diagnosis which is often missed in the
emergency room.Other tests that may be helpful in ruling
out other conditions are:
o Electromyography (EMG) or antibody studies may
help with the exclusion of myasthenia gravis and
Lambert-Eaton myasthenic syndrome (LEMS).
o Collection of cerebrospinal fluid (CSF) protein and
blood assist with the exclusion of Guillan-Barre
syndrome and stroke.
o Detailed physical examination of the patient for any
rash or tick presence helps with the exclusion of any
tick transmitted tick paralysis.
Treatment:
In the case of a diagnosis or suspicion of botulism, patients
should be hospitalized immediately, even if the diagnosis
and/or tests are pending. If botulism is suspected, patients
should be treated immediately with antitoxin therapy in order
to reduce mortality. Immediate intubation is also highly
recommended, as respiratory failure is the primary cause of
death from botulism.
Mortality from botulism ranges from less than 5 percent to 8
percent.
Prevention:
 Boil home-canned foods 10 minutes
 Follow USDA instructions on home- canning
 Restrict honey from < 1 year old
 Seek medical care for wounds
 Avoid injectable street drugs
Conclusions:
Regardless of the mode of exposure, botulinum toxins
produce a distinctive syndrome of cranial nerve palsies that
may be followed by descending flaccid paralysis. Effective
treatment depends on provision of intensive care and rapid
administration of botulinum antitoxin based on clinical
presentation, because laboratory diagnosis is time-consuming.
Free expert clinical consultation and antitoxin can be rapidly
obtained from the CDC by contacting public health authorities
.Rapid diagnostics and more-advanced specific therapy may
accelerate diagnosis and facilitate treatment.
References:
 Clinical Infectious Diseases by Jeremy Sobel, Volume
41, Issue 8, 15 October 2005, Pages 1167–1173.
Botulism | Clinical Infectious Diseases | Oxford
Academic (oup.com)
 A. Harris, in Encyclopedia of Food and Health, 2016.
Clostridium botulinum - an overview | ScienceDirect
Topics
 Peck MW (2009). Biology and Genomic Analysis
of Clostridium botulinum. Advances in Microbial
Physiology. Advances in Microbial Physiology.
Biology and Genomic Analysis of Clostridium botulinum
- ScienceDirect