1. BIOTERRORISM: is it a real threat? Zofia Piotrowska-Seget Department of Microbiology University of Silesia

2. Bioterrorism Microorganisms that infect and grow in the target host producing a clinical disease that kills or incapacitates the targeted host. Biologically Derived Bioactive Substances (BDBS) products of metabolism (usually, but not always, of microbial origin). These include biological toxins, as well as substances that interfere with normal behavior, such as hormones, neuropeptides and cytokine. The unlawful use of microorganisms or toxins derived from living organisms to produce death or disease. The act is intended to create fear and/or intimidate governments or societies in the pursuit of political, religious, or ideological goals.

3. Targets of bioterroristic attack people animals plant food water

4. History of bioterrorism Ancient times Poisoned arrows Solon poisoned the water supply with hellebore (skunk cabbage), an herb purgative (battle of Krissa). 184 B.C. During the naval battle against King Eumenes of Pergamon, Hannibal's forces hurled pots filled with serpents upon enemy decks. //www.zoetecnocampo.com/Documentos/cornezuelo 1346 During the siege of Kaffa, the Tartar army hurled its plague-ridden dead over the walls of the city. The defenders were forced to surrender.

5. 15th Century During Pizarro's conquest of South America, he improved his chances of victory by presenting to the natives, as gifts, clothing laden with the variola virus 1763 Captain Ecuyer of the Royal Americans, under the guise of friendship, presented to the native Americans blankets and the handkerchiefs contaminated with smallpox History of bioterrorism http://faculty1.coloradocollege.edu/~cneel/h y105/Black_.jpg 1915 Dr. Anton Dilger, grew cultures of Pseudomonas mallei supplied by the German government, in his home. The agents and an inoculation device were given to sympathetic dockworkers in Baltimore to infect 3000 horses, mules, and cattle, destined for the Allied troops in Europe http://www.saskschools.ca/curr_content/native30/photo s/bbTrade.gif

6. History of bioterrorism World War I Germany used Burkholderia mallei to infect horses and mules on the Eastern Front Due to contact with the infected animals people also became ill 1914-1917 Germans attempted to spread cholera in Italy, plague in St. Petersburg, and biological bombs over Britain http://www.appaloosa.com.br/mormo/ http://www.fotoinf o.pl/fil_41e.jpg http://www.first-world-war.com/images/dhm1361.jpg

7. International law Protocol for the prohibition of the use in war of asphyxiating, poisonous or other gases and of bacteriological methods of warfare on June 17, 1925, This protocol does not prevent the stockpile or development of such weapons and many state parties, reserved the right to strike against non-member states using such weapons or to retaliate when such weapons were used in a first strike, Geneva Protocol is the first multilateral agreement that extends the prohibition of chemical agents to biological agents. http://images.google.pl/imgres?imgurl=http://w ww.pilsudski.org/images/Sosnkowski.jpg

8. Unit 731 1936 Unit 731 was formed. Ishii constructed a 150 building complex just outside of Harbin, Manchuria for experimental purposes. Over 9000 test cases eventually died there. Ishii tested biological warfare on the Chinese, soldiers and civilians. Tens of thousands died as a result of plague, cholera, anthrax, etc., http://people.bu.edu/wwildman/WeirdWildWeb/courses/thth/project s/thth_projects_2003_parkeun.htm http://www.z-files.de/paranormal/ verschwoerung/ishii.jpg 1932 As Japanese troops invaded Manchuria, Shiro Ishii, a physician & army officer, began experiments on biological warfare. 1941 - plague was introduced by the Japanese in Suiyuan and Ninghsia provinces. A serious epidemic followed.

9. 1941 The British experimented with anthrax off the Scottish coast. Despite attempts to disinfect Gruinard Island, the spores left by the experiments kept the island for 48 years. The island was totaly disinfected in 1986 year. http://news.bbc.co.uk/1/hi/scotland/1457035.stm http://www.gifte.de/B-%20und%20C-Waffen/gruinard_island_bild01.htm II World War

10. Cold war Both USA and Soviet Union explored the use of hundreds of different bacteria, viruses, and biological toxins. Each program devised sophisticated ways to disperse these agents in fine-mist aerosols, to package them in bombs, and to launch them on missiles. In 1979, a rare outbreak of anthrax disease in the city of Sverdlovsk killed nearly 70 people. The Soviet government publicly blamed contaminated meat, but U.S. intelligence sources suspected the outbreak was linked to secret weapons work at a nearby army laboratory. 1942: The U.S. begins an offensive biological weapons program at Camp Detrick. After the war, efforts were continued at the renamed Fort Detrick and Pine Bluff.

11. 1969: President Richard Nixon ended the U.S. biological weapons program, and pledged the nation will never use biological weapons under any circumstances. The entire arsenal was destroyed by 1973, except for seed stocks held for research purposes. 1972: The Convention on the Prohibition of the Development, Production, and Stockpiling of Bacteriological (Biological) and Toxin Weapons and on Their Destruction is signed by more than 100 countries, going into effect in 1975. Signatories include Iraq and all permanent members of the United Nations Security Council. Cold war

12. Classification of biological agents Bacillus anthracis (anthrax) Yersinia pestis (plague) Variola major (smallpox) Francisella tularensis (tularemia) Botulin toxin (Clostridium botulinum) Viral hemmorrhagic fevers: Ebola, Marburg (filovirusy), Lassa (adenovirus) According to Centers for Disease Control and Prevention, Atlanta Category A Microorganisms that pose a special risk to national security can be easily disseminated or transmitted from person to person; result in high mortality rates and have the potential for major public health impact; might cause public panic and social disruption; require special action for public health preparedness. http:// www.bt.cdc.gov/agentlist-category.asp

13. Coxiella burnetii (Q fewer) Brucella (Brucellosis) Burkholderia mallei (Glanders) Staphylococcal enterotoxin B Epsilon toxin Clostridium perfringens Ricin toxin (Ricinus communis) Viral encephalitis (e.g Venezuelan equine encephalitis, eastern equine encephalitis) Food safety threats: Salmonella Shigella dysenteriae Escherichia coli O157:H7 Water safety threats: Vibrio cholerae Cryptosporidium parvum Category BCategory C virus Nipah virus Hanta yellow fever virus

14. Bacillus anthracis Robert Koch (1876 year) isolated and described this rod Aerobe, spore-forming bacterium : 3 – 8 μm 1 – 1,5 μm www.textbookofbacteriology.net

15.  Zoonotic disease in herbivores (e.g., sheep, goats, cattle) follows ingestion of spores in soil  Human infection typically acquired through contact with anthrax-infected animals or animal products or atypically through intentional exposure  Three clinical forms  Cutaneous  Inhalational  Gastrointestinal Anthrax: basics

16. Virulence factors pXO1pXO2 poly-D-glutamic acid capsule anthrax toxin composed of three proteins http://www.the- optimists.org.uk/authors/mark_g/graphics/full/anthrax-toxin.gif

17. Smallpox 1979 - eradication of smallpox Now the disease is eradicated after a successful worldwide vaccination program. Officially, the only remaining stocks of smallpox virus are held at the VECTOR institute in Koltisovo in Russia and at the Centre for Disease Control and Prevention in Georgia, USA Smallpox is a serious, contagious, and sometimes fatal infectious disease caused by virus Variola major Overall mortality rates were approximately 30% http://www.innovations- report.de/html/berichte/biowissenschaften_chemie

18. Botulin toxin (A) COOH SS HNHN H N 1 H N 2 NH 2 Neurotoxic protein produced by the bacterium Clostridium botulinum It is possibly the most acutely toxic substance known, with a lethal dose of about 200-300 pg/kg This toxin is rapidly destroyed by heat, such as in thorough cooking Zn 2+ Arnon S. et al. 2001. Botulinum toxin as a biological weapon.JAMA, 285:1059-1070.

19. Al-Hakam Missiles: 13 with botulin toxin 10 with aflatoxin 2 with anthrax spores Bombs: 100 with botulin 50 with anthrax spores 7 with aflatoxin IRAQ Zilinskas RA. Iraq's biological weapons: the past as future? JAMA.1999;278:418-424. In the early 1990's it was discovered that Iraq had produced 8,000 liters of anthrax spores. 8,000 liters is a sufficient amount capable of killing every man, woman, and child on earth.

20. Ebola virus Zoonotic virus 4 biotypes Ebola: Zair, Sudan, Reston, Ivory Cost Ebola haemorrhagic fever Symptoms: high fever, headache, abdominal pain, severe organ damage (especially the kidneys, spleen and liver) as a result of disseminated systemic necrosis. Among humans, the virus is transmitted by direct contact with infected body fluids or to a lesser extent, skin or mucus membrane contact. From CDC collection

21. Ricin toxin Lethal dose – 1 mg can kill adult The seeds from the castor bean plant, Ricinus communis, are poisonous to people, animals and insects Ricin toxin inhibits protein sythesis by specifically and irreversibly inactivating eukaryotic ribosomes. The symptoms of human poisoning begin within a few hours of ingestion. The symptoms are: abdominal pain vomiting diarrhea, sometimes bloody decrease in blood pressure

22.  Highly infectious; requiring only a few organisms to cause the desired effect (e.g. smallpox) or highly effective; requiring a small quantity of material to cause the desired effect (e.g. botox),  Efficiently dispersible, usually in the air; contagious or effective on contact,  Readily grown and produced in large quantities,  Stable in storage; preferably in a ready-to-deliver state,  Resistant enough to environmental conditions so as to remain infectious or operational long enough to affect the majority of the target, but not so persistent as to affect the occupying army. Features of the perfect biological weapon

23. Most agents relatively easy to produce – Availability of information on the Internet – Access to dual use equipment Relatively inexpensive cost of 50% casualty rate per km 2 conventional - $2,000 nuclear - $800 anthrax - $1 Many casualties but preserves structures Bioterrorism: a real threat

24. Dissemination may cover large area Difficult to detect release Symptoms occur days or weeks later Some have secondary spread Use can cause panic Users able to protect selves Users can escape before effect Bioterrorism: a real threat

25. Inglesby T.V et al. 1999. Anthrax as a biological weapon. Medical and public health management. JAMA, 281: 1735-1745. World Health Organization (WHO) expert committee estimated that casualties following the theoretical aircraft release of 50 kg of anthrax over a developed urban population of 5 million would be 250,000 - 100,000 of whom would be expected to die without treatment

26. Can we stop bioterrorism? Clarifying the cause of suspicious disease outbreaks Increasing cooperation among international agencies, Improving peaceful cooperation between scientists to make their activities more "transparent," Looking for bio-weapons specialists from the former Soviet Union or Iraq who disappear, stop publishing, or suddenly get rich, Controlling international trade in biological equipment useful for military and civilian purposes,