1. Clostridium botulinum and botulism

2. Introduction About 900's:  Certain foods caused typical poisoning.  Emperor Leo VI of Byzantium forbade the manufacture of blood sausages.

3. Introduction 1793: An outbreak caused by blood sausages was described in Wildbad, Germany

4. Introduction van Ermengem isolated an anaerobic bacterium from cured raw ham that had caused “Kerner’s Disease” in 23 people and killed 3. 1897

5. Other Findings killed a number of different experimental animals with the same signs as the disease in humans Extract from ham & a culture of m.o.

6. Another outbreak Caused by canned white beans The signs and symptoms were typical of botulism The toxin did not cross-react: type A Subsequently more toxin types found

7. How about the US? Between 1918-1922:  297 cases and 185 deaths  mainly in California  Prompted the cannery program that still exists today

8. Introduction 1936: C. botulinum type E was isolated  from smoked fish that caused botulism in the US and Russia

9. Introduction 1951: Wound botulism was described for the first time.

10. Categories of Human Botulism 1.Foodborne botulism 2.Infant botulism 3.Wound botulism 4.Adult infectious botulism

11. Categories of Human Botulism Infant botulism  It was first recognized in 1976.  This type of poisoning affects infants under the age of 12 month.  It is caused by the ingestion of C. botulinum spores.

12. Categories of Human Botulism Infant botulism  The spores colonize the intestinal tracts of infants, germinate, multiply, and produce neurotoxin.  The neurotoxin travels through the bloodstream to the central nervous system and causes flaccid paralysis.

13. Categories of Human Botulism Wound botulism  This illness results from the pathogen itself infecting a wound.  Foods are not the vehicle of transmission.  The microorganism produces the neurotoxin which is transmitted to other parts of the body via the blood.  Seen in intravenous drug users

14. Categories of Human Botulism Adult infectious botulism  This type of botulism resembles infant botulism, however, it affects adults.  C. botulinum colonizes the intestinal tract of adults and produces the toxin in vivo.  It has been thought to occur after antibiotic treatment depleted the indigenous intestinal flora.

15. Recorded Botulism Cases in California and in the US: 2002-2007 Botulism TypeCaliforniaU.S. (2007 not available) Food1980 Infant207435 Wound132155 AdultNot Avail. 4 http://ww2.cdph.ca.gov/data/statistics/Pages/CD_Tables.aspx http://www.cdc.gov/nationalsurveillance/botulism_surveillance.html

16. Classification of C. botulinum There are seven types of C. botulinum  A, B, C, D, E, F, and G  based on the serological specificity of the neurotoxin produced Types A, B, E, and, very rarely, F are associated with human botulism (foodborne, wound and infant types). Types C and D affect animals. Type G has not been linked to illness up to this date.

17. Characteristics of C. botulinum pH values for growth  Types A and proteolytic B pH 4.6-8.5  Minimum pH for E is:  6.2 at 5°C, and  5.4 at 30°C

18. Implicated Foods in Botulism  Any food that can support the growth of this pathogen or allow the germination of its spores and eventually toxin production can be associated with this illness.  Low acid foods (pH>4.6)  Home canned or preserved low acid vegetables  Garlic in soybean oil  Foil-wrapped baked potato  Commercial carrot juice held at ambient temperature

19. Implicated Foods in Botulism North American Indian specialties  fish and fish eggs  seal flippers Other implicated foods include luncheon meats, ham, sausage, smoked and salted fish, and lobster.

20. Illness & Causative Agent Botulism is a serious paralytic illness. It is caused by a nerve toxin that is produced by the bacterium. It is a rare illness. It is much feared.

21. Nature of Illness Intoxication Onset is about 18 – 36 hrs after ingestion of the food containing the neurotoxin. Symptoms vary from a mild to severe illness.

22. Clinical Symptoms Symptoms include:  neurological signs  blurred or double vision  difficulty in speaking or swallowing  fatigue  lack of muscle coordination, and  difficulties in breathing

23. Toxigenic Dose Few nanograms of C. botulinum neurotoxin can cause illness. The neurotoxin produced is probably the most toxic compound made by a biological system. About 1 oz. of this toxin can kill 200 million people. Fortunately, the incidence of the illness is low.

24. Mechanism of Toxin Neurotoxin 0 binds to neurons 0 internalized 0 prevents release of acetyl choline (neurotrasmitter)

25. C. botulinum Outbreak In 1994, in Oklahoma, a 47-year old man was hospitalized for symptoms of progressive dizziness, blurred vision, slurred speech difficulty swallowing, and nausea. Twenty-four hours earlier the patient had eaten some home canned green beans and beef and potato stew. Green beans tested negative, but stew was positive

26. C. botulinum Outbreak Apparently,  the stew was cooked,  covered tight,  left out for four days at room temperature, and  then eaten without reheating

27. Other recent cases: October 2004 Refrigerated clam chowder that was stored in the consumer’s home at room temperature Consumer tasted it, spit it out because it tasted bad She put it out for her chickens The next morning the chickens were dead

28. Other recent cases: June 2006 A truck driver picked up a load of pasteurized refrigerated carrot juice in the Bakersfield area He kept a case of it, and left one bottle in the cab of his truck where he was sleeping He subsequently drank it and developed botulism signs and symptoms Other bottles from the case that had been refrigerated were negative

29. Prevention Assurance of destruction or inhibition of C. botulinum. Keep susceptible foods out of the temperature danger zone (4.4 – 60°C or 40 – 140°F). Botulinum toxin is destroyed by heating at 80°C for 30 min or boiling for a few minutes. Thus re-heating foods properly can be a controlling factor.

30. Antitoxin Produced in horses by injecting them with gradually increasing levels of toxin type A To make a bivalent antitoxin, the horse is then injected with toxin type B, starting with extremely low levels Plasma is subjected to purification steps such as enzyme treatment and ammonium sulfate precipitation Time required is a matter of months

31. Antitoxin Distributed by CDC In California, local health jurisdictions can contact the DCDC duty officer for release of antitoxin (see flow chart) HHS awarded contract to Cangene Corp. of Canada for 200,000 doses of heptavalent botulism antitoxin to be delivered to SNS starting in 2007 as part of Project Bioshield

32. Detection of Toxin Toxin can be detected and typed by mouse inoculation. Number of tests depends on how many mice the lab has available. An ELISA method has been developed for detection of toxin. Used by LRN labs Assumption: Active toxin will kill mouse

33. Detection of Toxin Department of Defense, USDA, and Food and Drug Lab Branch use ECL (electrochemiluminescence) with which they can do 400-500 samples per day CDC has developed a TRF (Time-resolved fluorescence) test that is also high capacity (based on 96-well plate).