Clostridium botulinium(Microbiology)

clostridium

1. CLOSTRIDIUM BOTULINIUM By Caroline Karunya Ponnarasi Kanagaraj TSMU-2 nd year group-04

2. CLOSTRIDIUM-introduction Clostridia : are strictly anaerobic to aerotolerant sporeforming bacilli found in soil as well as in normal intestinal flora of man and animals. There are both gram-positive and gram-negative species, although the majority of isolates are gram-positive. Exotoxin(s ) play an important role in disease pathogenesis . motile -- peritrichous flagella (exception: C. perfringens — nonmotile )   the sporangia– swollen typical clinical symptoms

3. C.botulinum Classification: 1.Domain –bacteria 2. Division – firmicutes 3. Class –clostridia 4. Order – clostridiales 5. Family – closridiacae 6. Genus –clostridium 7. Species - botuilinium

4. C.botulinum Characteristics: Gram positive rod Subterminal endospore Noncapsule Obligate anaerobe Morphology: able to produce the neurotoxin during sporulation, which can only happen in an anaerobic environment.   is a  lipase  negative microorganism that grows between  pH  of 4.8 and 7 and it can't use  lactose  as a primary carbon source Spores of the organism are highly resistant to heat, withstanding 100 °C for several hours.

5. TOXIN Virulence factor — botulinum toxin neurotoxin relatively heat-labile and resistant to protease types: A , B , C, D, E , F, G the most potent toxic material known potassium cyanide(KCN) mechanism of action Toxin → gut → blood → cholinergic synapses → block the release of exciting neurotransmitter, e.g., acetylcholine → flaccid paralysis 10,000 times

6. MECHANISM OF TOXIN. 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

7. flaccid paralysis

8. Pathogenesis Disease—Botulism from Latin botulus , "sausage" There are three forms: adult botulism , caused by ingestion of preformed toxin in food; infant botulism , in which the organism replicates and secretes toxin in the intestinal tract; wound botulism , in which the organism replicates in the wound and secretes toxin. Toxin binds to neuromuscular junctions of parasympathetic nerves and interferes with acetylcholine release, causing flaccid muscle paralysis.

9. CLNICAL MANIFESTATIONS Adult botulism: flaccid paralysis: double vision, dysphagia, difficulty in breathing & speaking ,rare gastrointestinal symptoms .cause of death: respiratory failure Infant botulism: manifestation : constipation, poor feeding, difficulty in sucking and swallowing, weak cry, loss of head control. Floppy baby prevention: free of honey

10. Patient at rest, bilateral mild ptosis, disconjugate gaze, symmetric facial muscles. Requested to perform max. smile . Ptosis, disconjugate gaze, mild asymmetric smile.

11. . Wound Botulism from a heroin user

12. Epidemiology U.S. incidence < 200 annual cases of all forms Approx 9 annual outbreaks of food-borne median of 24 cases Recent trend toward restaurant rather than home-preserved foods All ages and genders equally susceptible Mortality 25% prior to 1960 6% during 1990’s I

13. Epidemiology Incubation period Depends on inoculated dose Inhalational 12-18 hours in primate studies 72 hours in 3 known inhalational cases True incubation period is unknown Foodborne 6 hours to 8 days Wound 7.5 days (range 4-18 days) after injury

14. DIAGONOSIS Clinical diagnosis Diagnostic tests help confirm Toxin neutralization mouse bioassay Serum, stool, or suspect foods Infant botulism C botulinum organism or toxin in feces samples for diagnostic testing Serum, vomit, gastric aspirate, suspect food, stool Collect serum before antitoxin given

15. DIAGONOSIS Confirmation Takes 1-4 days Available only at CDC and state public health labs Mouse Bioassay Type-specific antitoxin protects vs. toxin in sample The assay can detect at minimal 0.03ng of toxin. Culture Fecal and gastric specimens cultured anaerobically Results in 7 to 10 days Unique features to help in diagnosis Disproportionate cranial nerve palsies More hyptonia in facial muscles than below neck Lack of sensory changes

16. 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 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%)

17. PREVENTION Natural disease 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 Vaccine Botulinum pentavalent toxoid Not available to general public In use for laboratory workers, military Protects vs. types A-E Long-lasting immunity Prohibits future therapeutic use of toxin

18. <<<QUESTIONS PLS>>>