CBW CH 339 K CBW An Ancient Tradition

CBW CH 339 K

CBW - An Ancient Tradition Sixth Century B. C. 431 -404 B. C. Fourth Century B. C. Circa 200 B. C. 187 B. C. 7 th Century ~ 1040 1347 1672 1767 1855 Assyrians reportedly used ergot fungus (Claviceps purpurea) to poison their enemy’s water wells Spartan armies use sulfur and toxic arsenic smoke during Peloponnesian War Chinese engineers use arsenic against underground sappers. Officers in Hannibal’s army adulterate the wine of African rebels with mandrake, which contains belladonna alkaloids causing hallucinations. Ambraciots (Greece) employ irritating smoke against Roman soldiers The Byzantine architect, Callinicus (“Kallinikos”), invents the first liquid incendiary— “Greek Fire. ” Scottish king poisons wine using a belladonna-like (“sleepy nightshade”) herb and gives to Norwegian enemies as “provisions” under pretense of surrender. Scots then slaughter the incapacitated Norwegians. Mongolians lay siege to Kaffa (in modern Ukraine) and throw corpses over city walls to spread bubonic plague. Bishop of Münster attempted the use of atropine-like drug in grenades in siege against city of Groningen. Attack backfires. British plot to supply cloths from a smallpox hospital ward to American Indian tribes in hopes of spreading disease. Sir Lyon Playfair suggests using cyanide-containing chemicals against Russian troops during Crimean War.

Fritz Haber • 1868 – 1934 • 1918 Nobel for synthesis of ammonia (Haber process) • Father of modern chemical warfare (probably has a plaque on his office door in Hell)

Haber’s Law Where • W = Haber Product, also known as lethality index • C = Concentration of the agent in the air • t = time of exposure The lower the Haber Product, the more lethal the agent Lower concentration for a longer time will yield the same effects as a larger concentration over a shorter exposure

Beginnings • The large-scale use of poison gas in modern warfare originated during WWI in the battle of Ypres, April 22, 1915 – 168 tons of Cl 2 were released from cylinders along a 4 mile front – 5700+ cylinders of gas, weighing 90 lbs. each – 6000+ French troops were killed within 10 minutes – Due to the vagaries of wind and the crude delivery method, a fair number of Germans were also killed or wounded • 1. 3 million casualties, 91, 000 fatalities were attributed to the use of phosgene, chlorine, and mustard gas during the course of the war. • Including my grand uncle Bill

Modern Chemical Agents • Early Agents (WW 1) – Inhalation (Choking) Agents • Chlorine • Chloropicrin • Phosgene – Vesicants • Mustards • Lewisite – Blood Agents (Metabolic Poisons) • Cyanide Post-WWI Agents – Nerve “gases”

Chlorine • • Thick greenish vapor First used by Germans in 1915 Dispensed from gas cylinders Caused massive casualties due to total lack of defense

Chlorine Effects (Initial) • • • Burning pain in the throat and eyes, Sensation of suffocation; Painin the chest, especially behind the sternum. Rapid, painful, difficult respiration with coughing Profuse lachrymation. Retching and vomiting, Severe headache Face violet red, ears and finger nails blue Respiration often over 40 and sometimes even 80 a minute Dyspnoea lasts about thirty-six hours, after which it gradually subsides, if death does not occur before. The patient then falls asleep and wakes up feeling much relieved.

Chlorine Effects (Pneumonial Stage) • A few hours later acute bronchitis or broncho-pneumonia develops. • Sputum viscid, yellow or greenish, and muco-purulent with occasional streaks of blood. • Respiration shallow and rapid, up to 80 a minute. • Rapid pulse • Fever as high as 104. • Possible delirium. • Pleurisy may occur • Empyema and gangrene of the lung may occur. • Most survive after long recovery

Use of Chlorine French cylinder attack on German trenches in Flanders, year unknown. Note dependence on wind direction for successful attack.

Chloropicrin CCl 3 NO 2 pungent, colorless, oily liquid oral–nasal irritation, coughing, and vomiting LC 50 = 20, 000+ mg-min/m 3 (i. e. usually sublethal) First used by Russians in 1916 in hand grenades Used as secondary agent to make targets remove their masks to vomit • Used today as a fumigant • • •

Phosgene COCl 2 Colorless gas, smells like new mown hay BP 8 o C LC 50 = 3200 mg-min/m 3 Common industrial chemical Reacts with amino groups on surface proteins of the lung alveolar cells, cross linking them • Shortness of breath, coughing, tight chest, fluid buildup in lungs causing asphyxia • Can be asymptomatic for up to 24 hours • • •

Synthesis CO + Cl 2 → COCl 2 (ΔHrxn = − 107. 6 k. J/mol) • Purified caron monoxide and chlorine gas are passed together through a porous bed of activated carbon • Reactor must e cooled to temps of 50 – 150 o C

Sulfur Mustards Bis (beta-chloroethyl) sulfide • • Vesicant (blister agent) Alkylating agent cross-links lipids, proteins, and DNA Not very volatile (610 mg/l at 20 o C) Strong smell like horseradish or garlic Persistant agent LD 50 (human) = 20 mg/kg pericutaneous LC 30 (human) = 900 mg-min/m 3 by inhalation Interestingly, the Canadians in 1942 determined that anthrax spores can survive in sulfur mustard

Action • Sulfur mustard readily loses Cl- to form a highly reactive cyclic sulphonium ion • This ion reacts with free amino groups on biomolecules – e. g. guanine nucleotides in DNA, lysine residues in proteins • Molecular disruption – particularly of DNA – leads to cell death. • Blistering of skin • Blindness from eye damage • Lung damage and asphyxia due to filling of lungs with fluid

Use of Mustard • WW 1 – both sides • Second Italo-Abysinnian War 1935 -36 (League of Nations fretted and did nothing) • Sino-Japanese War • Iran-Iraq War • American troops exposed to low levels during Gulf War 1

Mustard (continued) Iranian victim of Iraqi mustard attack, 1980’s Americans blinded by mustard, France, 1918

Production Pre-WWII mustard production plant at Edgewood Arsenal, MD

Synthesis 1) Lommel-Steinkopf method using 2 -chloroethanol 2) Modern method using ethylene oxide

Synthesis 3) Levinstein method using ethylene 4) Radical addition reaction using vinyl chloride

Lewisite Talked about before as inhibitor of PDH LC 50 = 1300 mg-min/m 3 Severe chemical burns Inhalation causes a burning pain, sneezing, coughing, vomiting, and pulmonary edema • AFAIK, never used (manufacturing process was being worked out when WW 1 ended) • Significant numbers of Chinese were killed in the latter years of the 20 th century from accidental exposure to stockpiles left by the Japanese • •

Cyanide (HCN or salt thereof) By inhalation or skin adsorption LC 50 = 3000 mg-min/m 3 First used by Austria in 1915 Used by Iraq against both Iran and Iraqi Kurds More commonly used for assassinations, murders, and executions (not easy to disseminate on battlefield) • Used by KGB to assassinate Stepan Bandera, Ukranian resistance leader, in Germany in 1959 • • •

Nerve Agents • We talked about these with respect to enzyme inhibitors • Organophosphates first developed by IG Farben in 1936 • Extensively produced by Germans but never used in WWII • Russians in Afghanistan (? ), Iraq versus Iran and versus Kurds • Tokyo subway attacks of 1995 by Aum Shinrikyo

Irreversible Inhibition at the NM Junction

Nerve Gases Sarin Tabun Soman VX Symptoms: Contraction of pupils, profuse salivation, convulsions, involuntary urination and defecation and eventual death by asphyxiation as control is lost over respiratory muscles.

Action of Nerve Agents

Common Nerve Agents

Toxicity of Nerve Agents Agent LD 50 (human) LC 50 Tabun (GA) 1000 mg 400 mg/min-m 3 Sarin (GB) 1700 mg 100 mg/min-m 3 Soman (GD) 50 mg 70 mg/min-m 3 VX 10 mg 50 mg/min-m 3

Use of nerve agents • Iraq produced (admitted values) – 3. 9 tons of VX – 795 tons of Sarin – 210 tons of Tabun – In addition to 2850 tons of good old mustard. • Sarin and Tabun were loaded into bombs and ballistic missile warheads.

Synthesis

Synthesis (cont. ) Alternative syntheses for sarin (GB) Top: binary synthesis (components used in inary munitions) Bottom: Di-di process using methyl phosphonyl difluoride and methyl phosphonyl dichloride

Production US GB (Sarin) Production Plant, Rocky Mountain Arsenal, CO (now closed)

Delivery BLU-80 B Bigeye Binary Chemical Munition Could deliver 180 lbs. VX

Delivery (cont. ) Honest John rocket warhead containing M 139 bomblets, each with about 1. 5 lbs of Sarin

Binary munitions • • Binary munitions contain reactants which produce active agent on mixing Can e shipped and stored with only one (or no) components present Final filling at staging are before use Greater safety in storage and transport

Delivery (cont. ) M 687 Binary 155 mm Projectile Forward chamber contains methyl phosphonyl difluoride Rear contains mixture of isopropyl alcohol and isopropyl amine Munition is shipped / stored with rear compartment empty for safe handling – filled for final use • Separation between chambers id broken upon firing, creating agent “on the fly. ” • •

Delivery (cont. ) • M 23 Chemical Land Mine • Last units were destroyed in 2008 • Charged with 10. 5 lbs VX

Nerve Gas Antidotes Atropine sulfate from Atropa belladonna (left) competes with acetylcholine for the receptor binding site Blocks ACH and offsets the effects of the nerve agent

Pralidoxime Hydrochloride (Pyridine Aldoxime Methyl Chloride) Usually given along with atropine Reversibly binds to the enzyme acetylcholinesterase, competing with organophosphate binding. Doesn’t inhibit acetylcholinesterase – reactivator.

2 -PAM Action

Antidote Delivery • US Mark 1 anti-nerve agent kit containing (top to bottom) atropine autoinjector, 2 -PAM autoinjector, and the carrying case. The two autoinjectors are held together by a safety clip which prevents accidental discharge

Chemical Agents - Persistence Classification of agents as “persisting, ” “transitional, ” or “volatile” based on dispersal times. Wind conditions, rainfall, and cloudiness will also impact persistence Persistent agents are best used for area denial; volatile agents if attacking troops are to follow through.

Biochemical Weapons • Split the difference between Chemical and Biological • Mostly Protein toxins

Amount of toxin needed to lethally expose humans in a 100 km 2 (38 mile 2)area. (from Spertzel RO, Wannemacher RW, Patrick WC, Linden CD, Franz DR. Technical Ramifications of Inclusion of Toxins in the Chemical Weapons Convention (CWC). Alexandria, Va: Defense Nuclear Agency; 1992: 18. DNA Technical Report 92 -116. )

LCt 50 for Selected Aerosols

Botulinum Toxin • Clostridium botulinum is a spore-forming, anaerobic bacterium found worldwide in soil. • Mix of 7 related toxins • LD 50 = 1 ng/kg subq, 3 ng/kg inhaled • Bind to receptor on nerve cell • Taken in by receptor-mediated endocytosis • Prevents release of acetylcholine at synaptic cleft • Death by flaccid paralysis, respiratory failure • 6000 liters of concentrate were produced at Al Hakkam in 1990 • Delivery by warhead or from underwing spray tanks • High terrorist potential but technically difficult to produce • High lethality, low persistence

Ricin and related toxins • We’ve discussed extensively • US and England produced and tested a ricin bomb in WWII, but never used. • Inhalation causes death through lung damage rather than the organ failure seen in internal exposures • Diffuse necrotizing pneumonia • Not as toxic as botulinum, but easy to produce in large quantities

Georgi Markov 1929 -1978 Assassinated by Bulgarian secret police in London ricin attack

• Modified umbrella used as delivery device • Pellet contained ricin (note size)

Ricinus communis – castor oil plant

Ricin B chain (the attachment bit)

Ricin uptake and release 1. 2. 3. 4. 5. 6. 7. endocytosis by coated pits and vesicles or, endocytosos by smooth pits and vesicles. The vesicles fuse with an endosome. Many ricin molecules are returned to the cell surface by exocytosis, or the vesicles may fuse to lysosomes where the ricin would be destroyed. If the ricin-containing vesicles fuse to the Trans Golgi Network, (TGN), thereís still a chance they may return to the cell surface. Toxic action will occur when RTA, aided by RTB, penetrates the TGN membrane and is liberated into the cytosol.

The possibility that ricin might be used as an asymmetric warfare weapon has not escaped the attention of the armed services. The last time I knew for sure, there were no effective antidotes.

Significant Terrorist Incidents Involving Chemical and Biological Agents Year 1946 1970 1972 1974 1980 1984 1991 1990 -1995 1998 2001 2003 -2004 Organization DIN ("Revenge" in Hebrew; also Dahm Y'Israel Nokeam, "Avenging Israel's Blood") (Germany) Weather Underground (United States) R. I. S. E (United States) Aliens of America (Alphabet Bomber) (United States) R. A. F. (Rote Armee Faktion) (Germany) Rajneshee Cult (United States) Minnesota Patriots Council (United States) Aum Shinrikyo (Japan) Aryan Nation (United States) The Covenant and the Sword (United States) Republic of Texas (United States) Unknown (United States) Fallen Angel (United States) Agents Arsenic Compounds Tried to obtain agents from Ft. Detrick by blackmailing a homosexual serviceman. Typhoid, diphtheria, dysentery, meningitis and several others to be delivered by aerosol. Nerve Agents Botulinum toxin Salmonella enterica serovar typhimurium Ricin Bacteria and viral agents, toxins, organophosphorus nerve agents. Yersinia pestis Ricin Bacterial and viral agents Bacillus anthracis Ricin

Biological Weapons • Bacterial – Anthrax – Plague – Tularemia – Brucellosis – Glanders – Q Fever • Viral – Smallpox – Encephalitis – Hemorrhagic Fevers

Biologicals – Modern Use • Japan dropped plague infected fleas over various parts of China in 1940 – 42 • Several thousand prisoners and POWs died from anthrax, botulism, brucellosis, cholera, dysentery, gas gangrene, meningococcal infection, and plague. • Iraq prepared the following prior to GW 1 – 166 bombs (100 botulinum toxin, 50 anthrax, 16 aflatoxin); – 25 Scud/Al Hussein missile warheads (13 botulinum toxin, 10 anthrax, 2 aflatoxin); – 122 -mm rockets filled with anthrax, botulinum toxin, and aflatoxin; – 422 spray tanks capable of being fitted to a fighter aircraft or remotely piloted aircraft, and spraying 2, 000 L over a target – artillery shells. • The Soviets had an enormous biological program up to the collapse of the USSR, despite signing the 1972 convention against BW • Richard Nixon terminated the American offensive BW program in 1969

Anthrax • Discussed when we talked about bacterial cell walls • Spore formers make good weapons • Produces two exotoxins – Lethal toxin – Edema toxin • Proteins have A and B chains • B chain is same for both – 83 kdal cell binding domain • 90 kdal Lethal factor • 89 kdal edema factor

Cutaneous anthrax • • Most common non-intended form Handling of infected animals or animal products < 1% lethal with treatment 1 -2 cm blister that ruptures to leave necrotic ulcer

Inhalation anthrax • Inhalation of spores • High lethality even with treatment • Frequently leads to meningitis Characteristic widening of mediastinum

GI anthrax • • • Consumption of infected meat Up to 50% lethality Nausea and vomiting (possibly bloody) Anorexia Bloody diarrhea Incubation usually 1 week or less

Spore formation Figure 1. Stages of sporulation. (A) Stage 0/I, (B) stage II, (C) stage III, (D) stage IV, (E) released spore. The hatched line around the spore in panels D and E is the coat. From: Cell. Mol. Life Sci. 59 (2002)

Bacillus anthracis

Production Do not try this at home!!!!! 1. It will not work as shown 2. You will die 3. Everyone you know will go to Gitmo

Delivery The M 33 500 -lb biological cluster bomb, which held 108 of the M 114 bombs. Photograph: Chemical and Biological Defense Command Historical Research and Response Team, Aberdeen Proving Ground, Md.

Plague • • Black Death killed millions in Europe in 1400’s Causative agent is Yersinia pestis Endemic in wild rodent populations Gram -, does not form spores

Bubonic Plague • Characterized by formation of buboes – erthymatous, tender swollen lymph nodes • Fever, headache, chills, nausea, vomiting, prostration, coma • 5 -15% will develop pneumonic plague • 11 -12% fatal in USA

Septicemic Plague • Fever, chills, vomiting, diarrhea, purpura, and necrosis of extremities • 33% mortality in USA • Can be primary, or secondary effect of bubonic form Parient recovering from septicemic plague – note necrosis of nose and fingers

Pneumonic plague • Highly infectious person -to-person through aerosol • Mortality 50% with treatment, essentially 100% without

Dispersal • Japanese found that bacterial dispersal was inefficient • Clay “bombs” containing infected fleas can be dropped with 80% flea survival • Each flea can pass > 20, 000 bacteria per feeding • In the late 80’s, Russians developed a dry, antibiotic-resistant weaponized form for direct dispersal • Direct dispersal (particles suspended in air) produces very high incidence of pneumonic disease

Tularemia – rabbit fever • • Francisella tularensis – gram negative Fever, localized skin lesions, lymphadenopathy, pneumonia High infectivity Pneumonic form has high mortality Low persistence (unlike anthrax) (Formerly) used in cluster bombs Vaccine available Possibly used by Hittites – 14 th century BC – sending infected sheep as gifts Cutaneous Tularemia Francisella inside a liver cell

Brucellosis • • • Brucella sp. – gram negative coccobacilli Mainly infects ruminants in nature Field tested 1944 -45, never used Development terminated in USA in 1967 Aerosols highly infectious

Q fever • • Coxiella burnettii Rickettsia, normal reservoir in livestock Extremely High infectivity (as low as 1 particle) Moderately lethal, but a good incapacitant

Virus weapons • Good points – Not readily treatable – High lethality in some cases – Easier to engineer • Bad points – Difficult to stabilize, store, and disperse – Many viruses can be stabilized using very high concentrations of trehalose to form a glasslike material

Smallpox • • • Russians produced in large quantities Infects only humans Highly infectious (10 -100 particles) 35% mortality Extinct (hopefully) in the wild Stocks at Ft. Detrick, CDC in Atlanta, and Vektor – and (I think) Porton Down

Other Virus Agents Venezuelan equine encephalitis virus Incubation: 1 to 5 days 10 to 100 organisms Length of illness: 1 to 2 weeks Incubation: 3 to 6 days Length of illness: 1 to 2 weeks 5% mortality rate No specific therapy exists Vaccine available Nausea, vomiting, cough, sore throat and diarrhea can follow Low mortality rate Yellow fever virus Sudden onset of fever, severe headache, and muscle pain 1 to 10 organisms Severe fever, headache, cough, nausea, vomiting, vascular complications (including easy bleeding, low blood pressure) No specific therapy exists Vaccine available

Hemorrhagic Fever Viruses • High lethality – Marburg 50% – Ebola 60 -90% depending on strain • Tie up medical resources • Generally non-contagious • Russians weaponized Marburg virus, probably others

Delivery British Canberra Bomber, outfitted for biological agent delivery (notice spray apparatus)

Particle Penetration - Definitions

Delivery – Criticality of Particle Size (microns) Degree of Infectivity

Target Penetration as a Function of Particle Size Ideal particle size 15 mm for maximum lung penetration

Maximizing particle “fluffiness” • Silica particles in mix minimize van der Waals contacts and prevent clumping • Important to keep dry • Powdered glass or similar materials can carry a charge and help disperse particles • Other tricks are no doubt locked in a safe somewhere

Effectiveness

Cost Effectiveness Modality Cost per km 2 destroyed Conventional $6000 Nuclear $2400 Chemical (Nerve Agent) $1800 Biological $3 Note: 1969 dollars not adjusted for inflation “Destroyed” is defined as 50% casualty rate

Effectiveness by Amount Needed

Effectiveness of Biological Agents by Infectivity

Recent Terrorist Uses of CBW Significant Terrorist Incidents Involving Chemical and Biological Agents Year Organization Agents 1946 DIN ("Revenge" in Hebrew; also Dahm Y'Israel Nokeam, "Avenging Israel's Blood") (Germany) Arsenic Compounds 1970 Weather Underground (United States) R. I. S. E (United States) Aliens of America (Alphabet Bomber) (United States) R. A. F. (Rote Armee Faktion) (Germany) Rajneshee Cult (United States) Tried to obtain agents from Ft. Detrick by blackmailing a homosexual serviceman. Typhoid, diphtheria, dysentery, meningitis and several others to be delivered by aerosol. Nerve Agents Minnesota Patriots Council (United States) Aum Shinrikyo (Japan) Ricin Aryan Nation (United States) The Covenant and the Sword (United States) Republic of Texas (United States) Unknown (United States) Fallen Angel (United States) Yersinia pestis 1972 1974 1980 1984 1991 1990 -1995 1998 2001 2003 -2004 Botulinum toxin Salmonella enterica serovar typhimurium Bacteria and viral agents, toxins, organophosphorus nerve agents. Ricin Bacterial and viral agents Bacillus anthracis Ricin

Just to Make You Feel Secure… Vozrozhdeniye Island Biological Test Facility in Uzbekistan was the CCCP’s primary biological weapons test site. It is/was located on an island in the Aral Sea.

What could go wrong? Thanks to the wonders of Soviet agricultural engineering… Aral Sea 1971 Aral Sea 2001