BIOTERRORISM

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BIOTERRORISM

• SATURDAY MORNING PHYSICS
• Philip Hanna, Ph.D.
• Department of Microbiology Immunology
• University of Michigan Medical School
• pchanna_at_umich.edu
• February 10, 2007

Anthrax Spore magnified 92,000X
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Topics
Hot Fall Fashions! 2005 anthrax scare Ravens
game, Baltimore, MD

• What is bioterrorism?
• What are the most suspected agents?
• Is the threat real? If so, how did this come to
  be?
• Countermeasure efforts example anthrax.
• Society Science some challenges.

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Hannas Video Store therory of Fear
Bioterrorism
In the HORROR section of Hollywood Video, I find
2 predominant sub-genera of films
Unstoppable Forces of Nature Monster Flicks
Unrepentant Evil Slasher Flicks
Nightmare on Elm Street, 1984
Alien, 1979
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Definition
Agent of smallpox the variola virus, two
victims

• Bioterrorism the malevolent use of bacteria,
  viruses or toxins against humans, animals or
  plants in attempt to cause harm and create fear

Quarantine posting, 1936
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Select Agent list includes

• Bacteria plague, anthrax, tularemia, Q-fever,
  etc.
• Viruses smallpox. Marburg, Ebola, Lassa, Denge,
  Monkeypox, etc.
• Microbial Toxins BoTox, Tetrodotoxin, Ricin,
  etc.
• Genetically engineered microbes

www.cdc.gov/od/sap/
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Why Germs?

• Undetectable by our 5 senses
• Relatively inexpensive lt600X less Chem,lt800X
  Nuc
• Relatively low-tech to make deliver
• Small amounts hard to trace
• Delay in symptoms anonymity
• Chance of mass casualties
• Chance of fear, panic and disruption
• Very expensive to treat the sick and to
  decontaminate the places

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Why Germs?

• Germs in our FOOD supply
• Germs in our WATER supply
• Germs in our AIR supply aerosols
• Germs in our MAIL?
• Other ways means?
• Some diseases are transmitted person-to-person
  (Smallpox, plague, Ebola). Others are not
  (anthrax, botulinum toxin).

Crop dusting plane
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Biologic weapons are not new
1364 The Tartars, frustrated after years of
laying siege to the Black Sea city of Kaffa,
catapulted plague victims over unassailable city
walls.
1763 Sir Jeffery Amherst, commander of British
Troops in America, sanctioned the use of
Smallpox-infected blankets as germ warfare
agents against the Native Americans, who were
highly susceptible to the deadly virus.
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More instances
Aum Shinrikyo, a Japanese cult, attempted
several unsuccessful biologic attacks with
anthrax botulinum toxin before releasing
sarin gas in the Tokyo subway in 1995.
1984 The Rajneeshees, a religious cult, in
attempt to influence a local election poisoned 10
restaurant salad bars with Salmonella sickening
more than 700.
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2001 Anthrax Attacks Spores in US Mail 22
cases 11 cutaneous 11 inhalational 5 deaths
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Big Biowarfare

• United States, England Soviet Union had large
  efforts, highly developed, technologically
  advanced, weaponized and deployable offensive
  Bioagent Programs 20th century.
• US, under President Nixon, unilaterally
  discontinues all its ChemBio programs in 1969.
• Global Ban Treaty signed by gt100 nations in 1975.
• The Soviets, treaty signers, cheated big until
  the collapse of the Soviet Union.
• Nations still believed to have programs /or
  stockpiles.
• Tracking expertise, the weaponers, still
  difficult.

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Soviet Facilities
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Kazakhstan Production Facility
Anthrax Milling and Drying Facility, Kazahkstan
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Production-scale fermenters
Aerosol test chamber
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The 1979 Sverdlovsk Anthrax Outbreak
Resulted in scores of cases, 67 deaths
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Soviet Program
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Smallpox

• Variola major variola minor viruses (orthopox).
• Humans are the only natural reservoir.
• Endemic in people of all lands until latter half
  of the 20th century - fully eradicated in 1977.
• Highly contagious aerosol, body fluids, clothes,
  bed linens
• 12-14 day incubation period
• Fever, malaise, vomiting, head backaches
• 2-4 days later, pustular rash
• 30 - 40 mortality
• Fever contagious

Communicable Patient Transport
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Smallpox vs. Chickenpox
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Botulinum Toxin

• Made by the bacterium Clostridium botulinum
• Most toxic substance on earth - food poisoning
  ( 0.1 ug lethal dose)
• Weaponizable and aerosolizable (air food
  supplies)
• BoTox also used to medically, anti-wrinkle
• Secreted protein neurotoxin causing flaccid
  paralysis
• Death due to asphyxiation

Cosmetic improvements are NOT bioterrorism
C. botulinum
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Plague

• The Black Death 1/4 Europe killed in the 1300s
  (appx 30 million).
• 19th 20th century -10 million deaths in India
• Caused by the bacterium Yersina pestis
• Naturally, a zoonosis (from animals)

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Plague Symptoms

• General malaise
• High fever
• Pain/tenderness lymph nodes
• Convulsions
• Shock
• Diffuse, hemorrhagic changes in skin cyanosis
  dark skin Black Death
• Bubonic vs. pneumonic

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Two Forms of Plague
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Anthrax Countermeasures
Bacillus anthracis
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The center of my solar system
The sun
Anthrax spore
The other center of my solar system
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Anthrax Advent of Modern Microbiology Proof of
the Germ Theory
Robert Koch (1876)
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Anthrax and Advances in Immunology
Ilya Metchnikov
Louis Pasteur
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Bacillus anthracis

• Gram-positive, spore-forming rod
• Identified by Robert Koch in 1876 and used to
  formulate Kochs Postulates
• Causative agent of anthrax

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Sources of anthrax
Until recently, human infections came from
handling or eating contaminated animal products
In developed countries, anthrax is now very rare
B. anthracis is known to be part of biowarfare
arsenals worldwide, and is an attractive option
for terrorists
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B. anthracis pathogenesis
Dixon, et al. NEJM, 1999
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Progression of Systemic Anthrax 0
2 3 4
5 6 Days
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B. anthracis genome

• Chromosome 5.23 MB 5508 genes
• pXO1 toxin plasmid
• 182 KB 217 genes
• pXO2 capsule plasmid
• 95 KB 113 genes

Which of these encodes the next vaccine or drug
target?
T. D. Read et al, Nature 2003
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APPROACH

• DNA microarray construction for RNA expression
  studies
• Mass Spectrometry-based shotgun proteomics

T. D. Read et al, Nature 2003
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Cluster analysis of growth phase-regulated B.
anthracis genes
500
000

• K-median cluster analysis
• -24 clusters of co-expressed genes
• -4719 genes scored
• -3518 with gt2-fold change
• -2090 gt2-fold 2consecutive time points
• Arranged in order by time of activation

2090 genes
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High-Throughput Proteomics MuDPIT
Sample The Anthrax Spore
Lysis
Digestion
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Theoretical proteome, B. anthracis spore
Good non-bias with Multi-Dimensional Protein
Identification MuDPIT
Actual proteome, B. anthracis spore
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B. anthracis morphologies

• Endospore
• Stable for decades
• Resistant to uv, heat, desiccation , antibiotics
• Metabolically dormant, yet very sensitive to
  germinants in environment
• Infectious particle

• Vegetative bacilli
• Rapid growth (doubling time 15-20 minutes)
• Non-infectious

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Science Society

• Microbiology New understanding of pathogenic
  mechanisms of Bioagents.
• Medicine Select Agent vaccines, antibiotics,
  antivirals, diagnostics, training.
• Public Health Communication, facilities,
  education
• Detection real-time, surface antigen PCR, new
• Intelligence Law Enforcement dialogue
• Law Policy Quarantine Laws?, enforced
  vaccination?, Duel Use Research?
• Fear Terror get a better feeling for what
  germs are and what they can and cannot do.

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THANK YOU!
Cutaneous anthrax