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CLOSTRIDIUM
BOTULINUM
B Y
M D . A S H FA Q A Z I Z
D E PA R T M E N T O F M I C R O B I O L O G Y
P R I M E A S I A U N I V E R S I T Y
INTRODUCTION
• Clostridium botulinum is a Gram-positive, rodshaped
bacterium that produces several toxins. 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.
MORPHOLOGICAL CHARACTERISTIC
• A gram-positive, anaerobic bacilli.
• Spore forming.
• Toxin forming.
• Heat sensitive.
• Prefers low acid environment.
• Cause Botulism disease.
MORPHOLOGICAL CHARACTERISTIC
• 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
MORPHOLOGICAL CHARACTERISTIC
VIRULENCE FACTOR
• Neurotoxin:
- 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 micro gram.
VIRULENCE FACTOR
• 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) connected by a disulfide
bond.
- The A fragment of the nicked toxin, becomes the most potent toxin
VIRULENCE FACTOR
• Mechanism of Botulinum toxin:
– Absorbed from the gut.
– Binds to receptors of presynaptic membranes of motor neurons of the
peripheral nervous system and cranial nerves.
– Proteolysis-by the light chain of botulinum toxin of the target SNARE
proteins in the neurons inhibits the release of acetylcholine at the
synapse, resulting in lack of muscle contraction and paralysis.
– SNARE proteins are-synaptobrevin, SNAP 25, syntaxin.
– Type A and E toxin cleaves-SNAP 25.
– Type B toxin cleaves synaptobrevin.
VIRULENCE FACTOR
• Mechanism of Botulinum toxin:
Figure: Mechanism A Figure: Mechanism B
PATHOGENESIS
• Transmitted in three ways:
– Food or water toxin contamination.
– Wound infected with C. Botulinum.
– Ingestion of C. botulinum.
• Most common contaminated food:
Vacuum packed, or canned alkaline food.
E.g. fish, green beans, any home-canned food.
• Foods eaten without cooking
PATHOGENESIS OF INFANT BOTULISM
• Cause :infection by C. botulinum.
• Age: 5 - 20 weeks of age.
• Characterized by constipation and weak sucking.
• It cause “sudden infant death syndrome-SIDS”.
• Found in the stool.
SYMPTOMS
• Appear in 18-24 hrs.
• Include :
-Blurry vision, double vision. -Thrombocytopenia.
-Dry mouth.
-Trouble swallowing.
-Trouble breathing.
-Muscle weakness.
-Constipation.
-Dropping eyelid.
-Irritability.
• Fever is not a symptom of botulism.
SIGNS OF FOOD-BORNE AND WOUND
BOTULISM
• Ventilatory (respiratory) problems.
• Eye muscle paresis/paralysis (extra ocular, eyelid).
• Dry mucous membranes in mouth/throat.
• Dilated, fixed pupils.
• Ataxia.
• Hypotension.
• Nystagmus.
• Decreased to absent deep tendon reflexes.
DIAGNOSIS
• Clinical diagnosis
• Diagnostic tests help confirm
– Toxin neutralization mouse bioassay
• Serum, stool, or suspect foods
– Infant botulism
• C. botulinum organism or toxin in feces
• Testing is done in Reference laboratories under Biosafety regulations
TREATMENT
• Passive immunization - equine antitoxin
– Antibodies to Types A, B and E toxins.
– Binds and inactivates circulating toxin.
– Stops further damage but doesn’t reverse.
• Heptavalent antitoxin
– Investigational.
– Effective against all toxins.
TREATMENT
• Antitoxin action
– Food-borne botulism
• Neutralizing antibody levels exceed toxin levels
• Single dose adequate
– Large exposure (e.g. biological weapon)
• can confirm adequacy of neutralization
– recheck toxin levels after treatment
• Antitoxin adverse effects
– Serum sickness (2-9%), anaphylaxis (2%)
THANK YOU

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Clostridium botulinum

  • 1. CLOSTRIDIUM BOTULINUM B Y M D . A S H FA Q A Z I Z D E PA R T M E N T O F M I C R O B I O L O G Y P R I M E A S I A U N I V E R S I T Y
  • 2. INTRODUCTION • Clostridium botulinum is a Gram-positive, rodshaped bacterium that produces several toxins. 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.
  • 3. MORPHOLOGICAL CHARACTERISTIC • A gram-positive, anaerobic bacilli. • Spore forming. • Toxin forming. • Heat sensitive. • Prefers low acid environment. • Cause Botulism disease.
  • 4. MORPHOLOGICAL CHARACTERISTIC • 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
  • 6. VIRULENCE FACTOR • Neurotoxin: - 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 micro gram.
  • 7. VIRULENCE FACTOR • 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) connected by a disulfide bond. - The A fragment of the nicked toxin, becomes the most potent toxin
  • 8. VIRULENCE FACTOR • Mechanism of Botulinum toxin: – Absorbed from the gut. – Binds to receptors of presynaptic membranes of motor neurons of the peripheral nervous system and cranial nerves. – Proteolysis-by the light chain of botulinum toxin of the target SNARE proteins in the neurons inhibits the release of acetylcholine at the synapse, resulting in lack of muscle contraction and paralysis. – SNARE proteins are-synaptobrevin, SNAP 25, syntaxin. – Type A and E toxin cleaves-SNAP 25. – Type B toxin cleaves synaptobrevin.
  • 9. VIRULENCE FACTOR • Mechanism of Botulinum toxin: Figure: Mechanism A Figure: Mechanism B
  • 10. PATHOGENESIS • Transmitted in three ways: – Food or water toxin contamination. – Wound infected with C. Botulinum. – Ingestion of C. botulinum. • Most common contaminated food: Vacuum packed, or canned alkaline food. E.g. fish, green beans, any home-canned food. • Foods eaten without cooking
  • 11. PATHOGENESIS OF INFANT BOTULISM • Cause :infection by C. botulinum. • Age: 5 - 20 weeks of age. • Characterized by constipation and weak sucking. • It cause “sudden infant death syndrome-SIDS”. • Found in the stool.
  • 12. SYMPTOMS • Appear in 18-24 hrs. • Include : -Blurry vision, double vision. -Thrombocytopenia. -Dry mouth. -Trouble swallowing. -Trouble breathing. -Muscle weakness. -Constipation. -Dropping eyelid. -Irritability. • Fever is not a symptom of botulism.
  • 13. SIGNS OF FOOD-BORNE AND WOUND BOTULISM • Ventilatory (respiratory) problems. • Eye muscle paresis/paralysis (extra ocular, eyelid). • Dry mucous membranes in mouth/throat. • Dilated, fixed pupils. • Ataxia. • Hypotension. • Nystagmus. • Decreased to absent deep tendon reflexes.
  • 14. DIAGNOSIS • Clinical diagnosis • Diagnostic tests help confirm – Toxin neutralization mouse bioassay • Serum, stool, or suspect foods – Infant botulism • C. botulinum organism or toxin in feces • Testing is done in Reference laboratories under Biosafety regulations
  • 15. TREATMENT • Passive immunization - equine antitoxin – Antibodies to Types A, B and E toxins. – Binds and inactivates circulating toxin. – Stops further damage but doesn’t reverse. • Heptavalent antitoxin – Investigational. – Effective against all toxins.
  • 16. TREATMENT • Antitoxin action – Food-borne botulism • Neutralizing antibody levels exceed toxin levels • Single dose adequate – Large exposure (e.g. biological weapon) • can confirm adequacy of neutralization – recheck toxin levels after treatment • Antitoxin adverse effects – Serum sickness (2-9%), anaphylaxis (2%)