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Biological Warfare Agents
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- 2. Several bacterial, viralagents and toxins- pose public health risk- bioterrorist attack1 14th century- siege of ukraine2 Fort Pitt, Ohio river valley3 Anthrax- 1979, Soviet Union Anthrax, botulinum and aflatoxin- 1995, Iraq 1. Bioterriorism: from threat to reality.Atlas RM Annu Rev Microbiol. 2002; 56():167-85. 2. Biological warfare. A historical perspective.Christopher GW, Cieslak TJ, Pavlin JA, Eitzen EM Jr. JAMA. 1997 Aug 6; 278(5):412-7. 3. Wheelis M. Biological warfare before 1914. In: Moon JE van Courtland., editor. Biological and toxin weapons: Research, development, and use from the middle ages to 1945. Vol. 1. Stockholm, Sweden: Stockholm International Peace Research Institute; 1991. pp. 8–34. 4/20/2015 2
- 3. 4. Why shouldwe be concerned about biological warfare?. Richard Danzig et al,. JAMA, Vol 278,No,5,pp. 431-432 Factors Easy delivery Low visibility, high potency Recipes – available on internet Extremely Low- technology methods concealment, transportatio n ,disseminatio n easy 4/20/2015 3
- 4. 4/20/2015 4 5. J PharmBioallied Sci. 2010 Jul-Sep; 2(3): 179–188.
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- 8. Disease Pathogen Abused AnthraxBacillus antracis (B) First World War Second World War Soviet Union, 1979 Japan, 1995 USA, 2001 Botulism Clostridium botulinum (T) – Plague Yersinia pestis (B) Fourteenth-century Europe Second World War Smallpox Variola major (V) Eighteenth-century N. America Tularemia Francisella tularensis (B) Second World War 6. The history of biological warfare. EMBO Rep. 2003 June; 4(Suppl 1): S47–S52 4/20/2015 8
- 9. Agent Infective Dose (Aerosol) Incubation Period Diagnostic Assay Chemotherap y Anthrax 8,000-50,000 spores 1-5 d Ag-ELISA Ciproflaxin Doxycycline Penicillin Plaque 100-500 organisms 2-3 d Ag-ELISA Chlorampheni col Q-fever 1-10 organisms 10-40 d ELISA Tetracycline Small pox Assumed low 10-100 org 7-17 d ELISA,PCR, Virus isolation Cidofovir 7. Clinical Recognition and management of patients exposed to biological warfare agents. JAMA, Vol,278,No5.1997.pp.399-411 4/20/2015 9
- 10. • A releaseof 50 kg agent in an area with population 5 million • Anthrax 250,000 cases -100,000 deaths Plague 150,000 cases -36,000 deaths Tularemia 250,000 cases -19,000 deaths 4/20/2015 10
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- 12. Gram-positive, Endospore-forming, rod-shaped width of 1–1.2µm and a length of 3–5µm Only obligate species on Bacillus 4/20/2015 12 6. The history of biological warfare. EMBO Rep. 2003 June; 4(Suppl 1): S47–S52
- 13. Cell bindingprotein- Protective Antigen(PA) Enzymes • Edema Factor (EF) • Lethal Factor (LF) 4/20/2015 13
- 14. 14 • 95% ofall cases globally • Incubation: 3-5 days (up to 12 days) • Spores enter skin through open wound or abrasion Large skin ulcer created • Fever and malaise 5% - 20% mortality • Untreated – septicemia and death. Cutaneous • Severe gastroenteritis • Incubation: 2-5 days after consumption of undercooked, contaminated meat • Case fatality rate: 25-75% Gastrointestinal • Incubation: 1-7 days • Phase 1: Nonspecific - Mild fever, malaise • Phase 2: Severe respiratory distress Cyanosis, death in 24-36 hours • Case fatality: 75-90% (untreated) Inhalational 4/20/2015
- 15. 15 Worst-case scenario (Office ofTechnology Assessment) • 50 kg of spores Urban area of 5 million 250,000 cases of anthrax 100,000 deaths • 100 kg of spores Upwind of Wash D.C. 130,000 to 3 million deaths 4/20/2015
- 16. 16 Vaccine – availablebut effectiveness unproven in humans (only monkeys) • 5-35% experience systemic side effects • No long-term side effects proven • Six shots plus annual booster required Penicillin • Has been the drug of choice • Some strains resistant to penicillin Ciprofloxacin • Chosen as treatment of choice in 2001 • No strains known to be resistant 4/20/2015
- 17. Clostridium botulinum- Anaerobic bacteria Neurotoxins –A to G Incubation: 24 to 36 hrs 0.001 pg/kg of body weight 4/20/2015 17
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- 19. Vaccine : •Every case of Foodborne botulism is treated as a public health emergency. If antitoxin is needed, it can be quickly delivered to a physician anywhere in the country. • Skin should be tested for hypersensitivity before equine antitoxin is given. Mortality : • Botulism can result in death due to respiratory failure. • In the last 50 years, patients who die from botulism have dropped from 50% to 8%. 4/20/2015 19
- 20. Extreme neuro-specificity: BoNTsare being exploited in the treatment of a myriad of neuromuscular disorders and for the removal of facial wrinkles (BOTOX). 4/20/2015 20
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- 23. Techniques Binary Biological Weapons Designer Genes Gene Therapy 8. Genetically EngineeredBioweapons: A New Breed of Weapons for Modern Warfare. Mackenzie Foley .Applied Sciences, Winter 2013 / March 10, 2013 4/20/2015 23
- 24. The unfortunatefact remain that humans are often the most sensitive or the only detectors of the biological attack.9 9. Department of the Army, Navy and the Air Force NATO Handbook on the medical aspects of NBC Defensive operations,1996. 4/20/2015 24
- 25. Not onlyhuman sample Powdery material, air/water samples Sample preparation- hours to days Sample collection, handling, transportation and preparation- vital Conventional culture procedures- some virus or bacteria • Minimum: 3-7 to 15 days • Skilled manpower • No real time 4/20/2015 25
- 26. Detection in the fieldwith limited or no instruments Accurate Rapid 4/20/2015 26
- 27. • Natural prevalenceof the disease • Becton Dickinson (USA) , Vitek (BioMe’rieux) and Microlog (USA) • Pure culture and trained manpower Biochemical test based assays • Luciferin- luciferase interaction • Quality control- bacterial contamination • ATP contamination from non-microbial source • Non-specific • First line defence Bioluminescence based detection 4/20/2015 27
- 28. 10. Immunoassay ofinfectious agents.Andreotti PE, Ludwig GV, Peruski AH, Tuite JJ, Morse SS, Peruski LF Jr Biotechniques. 2003 Oct; 35(4):850-9. 11. A review of molecular recognition technologies for detection of biological threat agents.Iqbal SS, Mayo MW, Bruno JG, Bronk BV, Batt CA, Chambers JP Biosens Bioelectron. 2000; 15(11-12):549-78. • ELISA based • Quality of antigen or antibody11 • Different substrate label i.e, fluroscent, chemiluminescent and different platforms like ELISA plate, Visual dot and lateral flow format • Only one agent at a time Antigen- Antibody10 • Q-PCR assays-probes for all the agents • Software: Monitors the progress and presence detected online on a monitor • Data transferred over long distance • Variation with nucleic acids, availability of starting material • Inhibitory substances, specificity and sensitivity of primers, probes and enzymes used Nucleic acid based 4/20/2015 28
- 29. DRDE, Gwalior- • Toxicologyand biochemical pharmacology • nanotechnology-based sensors • unmanned robot-operated aerial and ground vehicles fitted with NBC detection sensors HSADL, Bhopal- • animal diseases such as avian influenza, Nipah virus infection, rabbit haemorrhagic fever, and swine flu. 4/20/2015 29
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