ANTHRAX

• Document Type: FOIA
• Collection: FOIA Collection
• Document Number (FOIA) /ESDN (CREST): 06785544
• Release Decision: RIPPUB
• Original Classification: U
• Document Page Count: 30
• Document Creation Date: March 9, 2023
• Document Release Date: November 23, 2021
• Case Number: F-2019-00776
• Publication Date: November 6, 2018

Anthrax - Intellipedia
Doc ID: 6640699
Approved for Release: 2021/11/22 C06785544
(b)(3) 50 USC 3605
(U) Anthrax
From Intellipedia
I _1) This page may not comply with Intellipedia:Classification policy.
., .
classification should match the classification of the contents.
(U) Per ICD 710 Paragraph G, Foreign Disclosure & Release (FD&R) markings are
mandatory on classified intelligence information.
(U) Please add or correct the classification markings. Mark the front of all portions,
See the discussion page for details.
including paragraphs, section titles, tables, links, and image captions. The banner
�CON-11-DEN-T-1-A-141440FORN�
(U) Anthrax is a serious illness caused by
Bacillus anthracis, an encapsulated, spore-
forming, large, gram-positive, aerobic
(grows in the presence of air), non motile
(unable to move independently),
rod-shaped bacterium. It is primarily a
disease of plant-eating animals
(herbivores); cattle, sheep, goats, horses,
and swine are the usual hosts. The bacteria
grow within the host, and sporulation
occurs when the bacteria are exposed to
oxygen or adverse growing conditions.
Virulent strains of B. anthracis produce a
protective capsule, composed of poly-D
glutamic acid, and two protein exotoxins
(called the lethal and the edema toxins).
(U) Definition in the WMD Term
Handbook An infectious disease of cattle
and sheep caused by the bacterium
Bacillus anthracis. The disease can be
transmitted to human by infected aerosol
inhalation anthrax or the respiratory form
presents as a flu-like illness with fever,
fatigue, nonproductive cough and chest
discomfort all of which last 2 to 3 days. It
may then progress to pneumonia with
respiratory distress, shock, and meningitis
generally leading to death in 24 to 36
hours despite appropriate therapy. Person
to person spread does not occur. However
the disease can be transmitted through
contact of contaminated animal substances
such as hair, bones, or hides. Direct
contact of infected materials with the skin
(cutaneous form) leads to the development
of a painless ulcer and swollen lymph
nodes. Gastrointestinal anthrax occurs
after ingestion of contaminated poorly
Contents
� 1 Type
� 2 Names
� 3 Simulants
� 4 History
� 5 Description/Symptoms
� 5.1 Inhalational anthrax
� 5.2 Gastrointestinal anthrax
� 5.3 Cutaneous anthrax
� 5.4 Mortality rates
� 5.5 Animal Studies
� 6 Diagnostics
� 6.1 Obstacles to Diagnosis
� 6.2 Diagnosis Based on Exposure History and Signs of Disease
� 6.3 Technical Methods to Confirm Diagnosis
� 6.4 Diagnostic methods for Infected Animals
� 7 Properties/Persistence
� 7.1 Properties of Anthrax
� 7.2 Persistence of Anthrax
� 7.3 Effect of Ultraviolet Radiation
� 8 Mechanism of Exposure/Effects
� 8.1 Mode of Transmission
� 8.2 Route of Infection
� 8.3 Poisoning
� 8.4 Toxicity
� 9 Prevention/Treatment
� 9.1 Prevention
� 9.2 Vaccines
� 9.3 Inoculation Schedule and Resulting Protection
� 9.4 Sensitivities and Side Effects
� 9.5 Treatment
� 9.6 Under Battlefield Conditions
� 9.7 For Respiratory Anthrax
� 9.8 For Cutaneous, Intestinal, and Oropharyngeal Anthrax
1 of 30
Approved for Release: 2021/11/22 C06785544
11/6/2018 9:29 AM
Anthrax - Intellipedia
Approved for Release: 2021/11/22 C06785544
(b)(3) 50 USC 3605
Doc 1D: 6640699
cooked meat or bone meal. The
therapeutic agents of choice are
ciprofloxacin, doxycycline, and penicillin
plus streptomycin. Ciprofloxacin in
combination with anthrax vaccine
provides effective prophylaxis in case of
exposure.[']
Type
(U) Bacillus anthracis is a bacterium.[21
This bacterium is the causative agent of
anthrax. This organism has the ability to
remain viable for decades in the
environment, especially the soil, in the
form of spores. Spores are the most highly
infectious phase of the bacterial life
cycle.[3]
Names
Anthrax is also known as woolsorter's
disease, splenic fever, Siberian ulcer,
Siberian boil plague, rag-pickers disease,
Persian fever, milzbrand, malignant
pustule (pustula maligna), malignant
carbuncle, and charbon.[2]
Simulants
The behavior of Bacillus anihracis can be
simulated by Bacillus szibtilis var. niger
(Bacillus globigii), and Bacillus
thuringiensis.[4][5]
History
Anthrax has been traced back to the fifth
and sixth plagues of Egypt, around 1500
B.C. The anthrax disease, known during
that era as "black bane" or "Murrain,"
caused serious losses of cattle. In the early
1700s, anthrax first appeared in North
America in Louisiana. Cutaneous anthrax
later appeared itself among cowboys on
cattle ranches in Kentucky in 1824. In
1864, anthrax was linked as a disease
caused by microorganisms living in the
blood of infected animals.
During the 1930s, extensive research was
conducted in Germany, Russia, and Japan
� 9.9 Immunity
� 9.10 For Exposed Animals
� 10 Alternate Uses
� 10.1 Vaccine
� 10.2 Immune system
� 10.3 Inserting the vaccine
� 10.4 Results of the vaccine
� 11 Production/Storage
� 12 Pilot Scale Production
� 12.1 Interim Report 108
61 12.3 Laboratory requirements
� 12.4 Seed selection
� 12.5 Drying methods for seed stocks
� 12.6 Starter cultures
� 12.7 Media production
a 12.7.1 Technical report BWL 12
� 12.8 Fermentation
� 12.9 Centrifugation
� 12.10 Hazards of production and storage
� 13 Detection
� 13.1 Role of detection
� 13.2 Architectural hardware
� 13.2.1 Triggers
� 13.2.2 Collectors
� 13.2.3 Detectors
� 13.2.4 Identifiers
� 13.3 Point detectors
E 13.4 Mobile Labs
� 13.5 Stand-off detectors
� 14 Anthrax as a Biological Warfare Agent
� 14.1 US CBW Program Data
� 15 Weaponization/Dispersal
� 15.1 Weapon Types
� 15.2 Bomblets
E 15.3 Types of Bomblets
� 15.4 Line Source Weapons
E 15.5 Other Delivery Systems
� 15.6 Stabilization
� 15.7 Meteorological Conditions for Release
� 15.8 Aerosol Form
� 15.9 Dispersal Efficiency
� 15.10 Meteorological Conditions for Dispersal
� 16 Physical Protection/Decontamination
� 16.1 Protective Clothing
� 16.2 Decontamination Exposure
16.3 Methods of Decontamination
E 16.4 Chemicals Used for Decontamination
� 16.5 Medical Instrument Decontamination
� 16.6 Proof of Decontamination
� 17 Russian Anthrax Compendium
2 of 30
Approved for Release: 2021/11/22 C06785544
11/6/20189:29 AN
Anthrax - Intellipedia
Approved for Release: 2021/11/22 C06785544
(b)(3) 50 USC 3605
Doc ID: 6640699
toward the use of anthrax as a biological
weapon. During World War II, several
countries produced anthrax, yet Japan was
the only country to use it as a biological
warfare agent (Unit 731). Japan produced
mass quantities of anthrax from 1939 until
1945.
� 18 Anthrax and Botulinum Handbook, 1993
� 19 (U) Non-IC Resources
� 20 References
Anthrax has surfaced in the news in the post-World War II era. In support of the
Biological Weapons Convention, the United States announced in 1969 that it would
destroy all stockpiles of biological agents and weapons, including anthrax. Also, two anthrax epidemics occurred in the
1970s. The first occurred among cattle in Rhodesia (now called Zimbabwe), and the second epidemic occurred in
Sverdlovsk. This outbreak resulted from an accidental release of anthrax spores from a production facility known locally as
Cantonment 19. In 1980, it was reported that Iranian prisoners of war died while undergoing anthrax testing. In the 1990s,
North Korea allegedly tested anthrax and other bioagents. In 1995, it was discovered that Iraq had conducted weapons trials
using anthrax as the fill agent.
Intelligence History Portal
For more information, see History of Anthrax
Description/Symptoms
B. anthracis bacteria are encapsulated, spore-forming, large, gram-positive, aerobic (grows in the presence of air), nonmotile
(unable to move independently), and rod-shaped. It is primarily a disease of plant-eating animals; cattle and sheep are
common hosts.
B. anthracis spores are extremely resistant to environmental factors. They can remain viable for several decades under
suitable environmental conditions. There are three forms of anthrax: inhalational, gastrointestinal, and cutaneous. The
incubation period for anthrax is 1 to 7 days, with most cases occurring within 2 days of exposure. The infection usually lasts
from 3 to 5 days. Each form of infection is unique with its own characteristic symptoms.
inhalational anthrax is the manifestation of the disease likely to be expected in biological warfare. The symptoms of
inhalational anthrax may vary.
sudden death is common, due to blood poisoning (septicemia) or inflammation of the lymph nodes
ymp a enitis). If the dose is small and the particle size is large there are usually notable
symptoms. Early symptoms include malaise (discomfort), fatigue, 'ever, ana an upper reSpiratory infection. These symptoms
are followed by the sudden onset of respiratory distress, which is followed by shock; death occurs within 24 to 36 hours.
Death occurs in 95 to 100 percent of untreated cases.
Gastrointestinal anthrax shows symptoms of abdominal distress followed by bloody stools, vomiting, fever, and signs of
septicemia. Death occurs in about 50 percent of all cases.
Cutaneous anthrax, also referred to as malignant pustule, malignant carbuncle woolsorter's disease, or rag-picker's disease,
develops when exposed abraded skin is infected with the bacteria. First, the skin itches, and within 2 to 5 days a skin ulcer
(lesion) forms. Later the lesion turns into a large black scab. Untreated infections can spread to regional lymph nodes,
causing blood poisoning. Meningitis can also occur. Approximately 5 to 20 percent of untreated patients develop septicemia
and generalized infection. It must be emphasized that sudden unexpected collapse and death are the most characteristic
indication of anthrax infections, especially in the inhalation form of the disease.
B. anthracis is found in soil (particularly dry soil) as a resistant spore that may persist for years under suitable environmental
conditions. Spores vegetate in the soil when the pH and temperature conditions are favorable. The bacteria are found most
commonly in areas with somewhat neutral soil (pH 6 to 8.5) and during periods of both drought and flooding. Flooding
allows the bacteria to accumulate at the ground surface in low-lying areas. Subsequent drought affords conditions for
exposure of the spores. The areas of the world where anthrax is endemic (prevalent) in animals are the Middle East, Africa,
and Central and South America. The spores are very resistant to heat, disinfectants, sunlight, and other environmental
factors. When the spores are inhaled, they convert to the vegetative form, establish an infection, and, as they multiply in the
host, produce highly lethal toxins. There are three primary classifications of infection by B. anthracis: cutaneous,
3 of 30 11/6/2018 9:29 AM
Approved for Release: 2021/11/22 C06785544
Anthrax - Intellipedia
Doc ID: 6640699
Approved for Release: 2021/11/22 C06785544
(b)(3) 50 USC 3605
gastrointestinal, inhalational. The incubation period of anthrax is 1 to 7 days, with most cases occurring within 2 days of
exposure. The infection usually lasts from 3 to 5 days. Each form of infection is unique with its own characteristic
symptoms.[61[7][81
Inhalational anthrax
When the dose of inhalational anthrax is large and the particle size is small
death is expected once symptoms occur. Because the average anthrax spore is 2.6 microns, it is small enough to
pass through the upper respiratory tract and penetrate the lungs. Anthrax spores are transported by host cellular immune
defense processes. This causes septicemia (blood poisoning) or hemorrhagic inflammation of the lymph nodes
(lymphadenitis). This is the type of anthrax expected to occur in biological warfare. After a short period of moderate general
disability and the rapid onset of septicemia death occurs.[9]
When the patient is infected with a small dose and a large particle size most of the particles settle in (b)(1)
the upper respiratory tract, such as the nasal passages and upper lungs, and do not reach the alveoli. The incubation period is
1 to 6 days. These patients have a prolonged course of prominent bronchial symptoms, which occur in two stages because
the particles clump together and cannot pass easily into the bloodstream. In the early stage of large-particle, low-dose
inhalation anthrax, the symptoms are mild, nonspecific, and characterized by insidious onset. Symptoms include mild fever,
discomfort (malaise), fatigue, muscle pain (myalgia), a nonproductive cough, and frequently a sensation of precordial
oppression (a feeling of heaviness in the chest). This initial stage typically lasts for several days. The patient's clinical
condition may improve slightly toward the end of this stage. The second stage of inhalational anthrax is acute toxemia. It
advances very rapidly and is associated with a massive invasion of the organism throughout the body. It develops with acute
difficult breathing (dyspnea) and bluish skin coloring (cyanosis). The patient appears moribund (unable to move), with
accelerated pulse and respiration. The body temperature, although usually elevated to 102�F or more, may suddenly become
subnormal because of shock. Harsh breathing (stridor) is common, and chest examination discloses crepitant rales (abnormal
crackling sound) associated with fluid in the lungs. The average duration of this acute stage is less than 24 hours,
terminating in death. Consciousness is usually maintained until death, except in the case of meningitis (which occurs in 50
percent of the cases), when disorientation and coma occur.[10][i I
Gastrointestinal anthrax
The least common, naturally occurring form of anthrax is gastrointestinal anthrax. It may develop secondary to a primary
lesion in the region of the mouth, or it may result from the ingestion of large numbers of spores in uncooked or undercooked
infected food (primarily meat). The incubation period is from 2-5 days. In the early stages of gastrointestinal anthrax, the
symptoms are mild and nonspecific. This early stage is followed by a very rapid onset of the advanced disease, which is
associated with a massive invasion of the organism throughout the body. Symptoms of acute gastrointestinal anthrax are
severe protracted vomiting, fever, signs of blood poisoning, and bloody diarrhea. The latter is relatively uncommon because
intestinal obstructions rapidly develop (adynamic ileus) with symptoms of fulminating, acute, generalized inflammation of
the abdominal cavity memliranes (peritonitis). No significant involvement of the membrane that lines the abdominal cavity
and contains the organs of the abdominal cavity (peritoneum) actually occurs, although a characteristic hemorrhagic
inflammation of the lymph nodes (lymphadenitis) in the peritoneal fold encircling the small intestines (mesentery) is
invariably present.[I�1[111
Cutaneous anthrax
In cutaneous anthrax, also known as woolsorter's disease or rag-picker's disease, invasion occurs through either abraded skin
or through small breaks in the layers of cells forming the epidermis of the skin (epithelium). The incubation period is from I
to 5 days. The signs start with irritated itching where the skin was exposed to the agent. Within a few hours, the affected area
appears as a small, red discolored spot or patch on the skin (macule). It progresses over a period of several hours to a firm,
red elevated area of skin that is solid and circumscribed (papule). It proceeds to enlarge into a skin ulcer (vesicle) with
surrounding fluid buildup (edema). The vesicle formation may be single or multiple with rings of satellite vesicles
surrounding the central lesion. The known symptoms for the early stages of cutaneous anthrax are discomfort (malaise),
fever, headache, and general exhaustion (prostration). In 2 to 3 days, the vesicle becomes hemorrhagic and is surrounded by
a deep red zone of hardened tissue (induration and erythema). The amount of local fluid build-up (edema) is variable and
4 of 30 11/6/2018 9:29 AN
Approved for Release: 2021/11/22 C06785544
Anthrax - Intellipedia
Doc ID: 6640699
Approved for Release: 2021/11/22 C06785544
(b)(3) 50 USC 3605
depends to a certain extent on the location of the lesion. In areas such as the skin of the eyelids, face, and neck, it can be
massive and present a serious complication. Under most circumstances, a zone of doughy edema surrounds the primary
lesion. During the active phase of the disease, most patients have significant fever, reaching maximum levels of 101�F. By
the sixth day, it manifests as a black scab, and fluid buildup subsides. At this point, the lesion and surrounding skin dies
(becomes necrotic). The wound is not painful, but the regional lymph nodes are tender. By this stage, blood poisoning
(septicemia) and meningitis can occur. The black scab can usually be removed after 10 to 14 days. The severity of the
symptoms depends on the stage of the disease and the dose (the number of organisms involved in the initial infection).
Approximately 5 to 20 percent of untreated patients will develop septicemia and generalized infection. More than 95 percent
of anthrax cases are cutaneous.[1�][121[II][131
Mortality rates
The mortality rate for inhalation anthrax is 95 to 100 percent in untreated cases, in intestinal anthrax, �50 percent and in
cutaneous, �20 to 30 percent in untreated cases. Mortality rates for oropharyngeal anthrax can be as high as 50 percent.
[10][14]
Animal Studies
In studies using laboratory grown-spores and exposure of monkeys to aerosols generated under controlled conditions, the
median lethal dose (LD 50) values were established.
How this data relates to humans is yet to be determined.1151
The symptoms for herbivores (cattle, sheep, swine, and goats) can be broken down into three forms: peracute (violent acute
symptoms), acute (rapid onset, severe symptoms, and a short course), and localized (restricted to a region). Peracute forms
are seen in cattle, sheep, and goats. These animals suffer from cerebral apoplexy (severe hemorrhage) and die, frequently
without showing any previous evidence of illness. In cattle, intermandibular swelling extending into the jugular furrow can
be noted. Severe difficult breathing (dyspnea), with head held low and outstretched, is common. Tremors and rapid
progression to lying down (recumbency) can occur. The acute form is most common in all species, except swine. The classic
symptoms of this type are fever, excitement, depression, stupor, spasms, respiratory or cardiac distress, convulsions, and
bloody discharges. In the localized form, swelling and circumscribed inflammation of the skin (carbuncles) in various parts
of the body occurs. In swine, anthrax usually is localized in the cervical lymph nodes, where it causes swelling and
hemorrhage. This form may progress to the acute stage or subside with recovery. In swine, death may occur from
161111[18]
suffocationJ
Diagnostics
Early diagnosis of anthrax is essential to prevent fatalities. A correct diagnosis is not easily accomplished, because the initial
symptoms of the disease are not unique to anthrax. The most critical aspect in making a diagnosis of any form of anthrax is a
strong suspicion of possible natural, occupational, or battlefield exposure.
Most of the technical methods for confirming a diagnosis of anthrax are useful only in a retrospective analysis, because the
patient often dies before conclusive results can be obtained. Available methods include DNA matching, visual examination
of smears and cultures, biochemical tests, immunoassays (detection of antibodies to specific antigens), and animal
inoculation and skin tests. Each method depends on identification of the disease-causing agent (pathogen) by matching
characteristics of a sample to a known pattern. The DNA-matching technique has the greatest potential to confirm a
diagnosis of anthrax before the patient dies, because the method is faster (and more specific) than other technical
approaches.
Obstacles to Diagnosis
Medical diagnosis is the process of applying scientific methods to establish the cause and nature of a person's illness. The
diagnosis is based on a health care practitioner's evaluation of the patient's subjective symptoms, the physical findings, and
(b)(1)
5 of 30 11/6/2018 9:29 AV
Approved for Release: 2021/11/22 C06785544
Anthrax - Intellipedia
Approved for Release: 2021/11/22 C06785544
(b)(3) 50 USC 3605
Doc ID: 6640699
the results of various laboratory tests, together with other appropriate diagnostic procedures. Early diagnosis of anthrax is
essential to prevent fatalities. Making a correct diagnosis is not easily accomplished because the initial symptoms of the
disease are not unique to anthrax. In order to determine the disease that is causing the symptoms, health practitioners often
begin by making a differential diagnosis that considers a number of diseases characterized by the patient's symptoms. For
example, the differential diagnosis of a respiratory (in the lungs) anthrax epidemic while still in the early stages of
nonspecific symptoms could be impeded by its similarities to a wide variety of viral, bacterial, and fungal infectious
diseases. The differential diagnosis for cutaneous (on the skin) anthrax could include tularemia, staphylococcal or
streptococcal disease, and orf (a viral disease of sheep and goats, transmissible to humans).[191 [20][21][13][15][14]
Diagnosis Based on Exposure History and Signs of Disease
The most critical aspect in making a diagnosis of any form of anthrax is a strong suspicion of possible natural, occupational
or battlefield exposure. For cutaneous mid naturally occurring respiratory anthrax, exposure most frequently arises from
proximity to infected animals or contaminated animal products. Under battlefield conditions, respiratory anthrax results
from exposure to a Bacillus anthracis-containing aerosol (i.e., weaponized anthrax). Gastrointestinal anthrax is exceedingly
difficult to diagnose because of the rarity of the disease. Diagnosis of this form of anthrax is usually based on evidence of an
outbreak due to ingestion of contaminated meat.[14]
Once the nature of the exposure is verified, physical signs of disease can be investigated to determine the form of anthrax
present. Cutaneous anthrax should be considered if the patient has a painless but itchy, raised reddened area of skin, often
with surrounding swelling due to fluid buildup in the tissues (edema), that develops into a black scar. The scar, together with
extensive edema, is strongly indicative of cutaneous anthrax.[I4]
If respiratory anthrax is suspected, X-rays should be performed to detect evidence of the widening of the mass of organs and
tissues separating the lungs (mediastinum). The widening is due to inflammation and fluid buildup, including blood, in the
mediastinum. The presence of meningitis, chest wall edema, and fluid buildup in the tissues surrounding the lungs also
points to respiratory anthraxf14][21][13]
Technical Methods to Confirm Diagnosis
Most of the technical methods for confirming a diagnosis of anthrax are useful only in a retrospective analysis, because the
patient usually is dead or dying before conclusive results can be obtained. (Between 20 and 100 percent of all untreated
forms of anthrax will result in fatalities.) Available methods include DNA matching, visual examination of smears and
cultures, biochemical tests, immunoassays (detection of antibodies to specific antigens), and animal inoculation and skin
tests. Each method depends on identifying the disease-causing agent (pathogen) by matching characteristics of a sample to a
known pattern.[22/[141123]
The DNA-matching technique has the greatest potential of confirming a diagnosis of anthrax before the patient dies, because
the method is faster than other technical approaches. Matching DNA sequences is a relatively new approach made possible
with the development of recombinant DNA techniques. DNA sequences are matched using the polymerase chain reaction
(PCR) blood test kits. PCR utilizes DNA probes to identify bacteria present in trace quantities in the blood. For anthrax, the
DNA probes would be fragments of Bacillus anthracis (B. anthracis) DNA marked with a radioactive isotope or fluorescent
dye. These labeled DNA fragments would pair up with DNA in a sample, if the sample contains DNA from the same
microorganism. The sensitivity of the test varies according to the length of the DNA fragment used for identification. The
longer the fragment, the more specific the identification of the bacteria; a long fragment might include 1,000 DNA base
pairs [23]
Traditional visual examination methods focus on the shape, color, and growth characteristics of bacteria found in samples of
infected blood, tissue, or body fluids. B. anthracis is a straight rod, 5 to 10 microns long, which does not move
spontaneously (nonmotile), grows in the presence of oxygen (aerobic), and produces spores. The microorganism is
gram-positive, which means that stained cells retain a bluish color after washing with organic solvents. Only those strains of
B. anthracis that have a protective outer coating or shell (capsule) and produce toxins are virulent (poisonous and
infectious). When grown in the laboratory, the organisms form long, curved chains and within 24 hours can produce large,
raised, opaque, grayish-white colonies with an irregular border. This irregularity will occasionally give individual colonies a
6 of 30 11/6/20189:29 AM
Approved for Release: 2021/11/22 C06785544
Anthrax - 1ntellipedia
Doc ID: 6640699
cornet tail appearance, which is quite characteristic.101
Approved for Release: 2021/11/22 C06785544
(b)(3) 50 USC 3605
The identification of bacterial microorganisms can be done by visual examination of smears (initial samples of infected
matter spread on a surface such as a glass slide) and cultures (large colonies of bacteria grown in the laboratory from an
initial sample). Smears require microscopic examination to view individual or small groups of microorganisms. Staining
techniques have been developed to aid in locating the bacteria. Smears are stained by Gram's method for determining shape
and staining properties and by the methods of McFadyean for demonstrating capsules. For blood smears, a Giemsa stain is
also satisfactory in demonstrating the presence of capsules.1I01
For cutaneous anthrax, samples are taken from the site of infection. Pathogenic strains of B. anthracis can usually be located
in the fluids found in cutaneous wounds, or in material scraped from the underside of a crust or scab of a skin wound. When
examined in smears from the blood or tissues, the organisms usually are found singly or in pairs with well-defined capsules.
The typical culturing method suspends samples in small quantities of saline solution that are inoculated directly onto agar
plates (a common solid culture medium using extracts from seaweed) or in broth (a liquid culture medium based on meat
extracts).[I�1
For respiratory anthrax, samples are taken from blood, sputum (material expelled by coughing), and body fluids drawn from
the lungs, spinal cord, or swollen lymph nodes. B. anthracis is readily detectable by blood culture with routine media.
Occasionally, bacilli may be identified in the centrifuged sediment of blood treated with 3 percent acetic acid solution and
stained with Wright's stain. Smears and cultures of pleural fluid (from the membrane surrounding the lungs) and
cerebrospinal fluid (from the brain and spinal cord) may also be positive for B. anthracis. Impression smears of mediastinal
lymph nodes (found near the lungs) and spleen from fatal cases are positive.[2I][13][24][25][261
For respiratory and intestinal anthrax, samples including specimens of vomitus and feces would most likely be contaminated
with other bacteria, making them unsuitable for examination or culturing. In these cases, inoculating animals can confirm a
diagnosis of anthrax. The best method of inoculating animals, preferably white mice or guinea pigs, with this material is by
simply scratching the skin. The anthrax bacillus can gain entrance through the abraded skin more easily than other
organisms that may be present. Prior to culturing or incubating such contaminated material, it may be heated to 65�C for 60
minutes, or to 80�C for 30 minutes. Another method is to suspend 0.1 ml of the specimen in 10 mL of 1 or 2 percent phenol
for one hour at room temperature. Both of these techniques take advantage of the extraordinary stability of anthrax
spores.[271
Animals also may be inoculated by injecting under the skin in the thigh uncontaminated initial or cultured samples
suspended in a small quantity of saline. After inoculation (using either contaminated or uncontaminated control samples), if
B. anthracis is present, death occurs in 36 to 72 hours with characteristic features of anthrax. Such features include edema at
the inoculation site; dark colored, uncoagulated blood; an enlarged, dark, easily broken spleen; and a congested, mahogany-
colored liver. The bacteria can be recovered readily from the blood of these animals.1281
In addition to examination of smears and cultures, various biochemical reactions with sugars can also be used to help
identify the organism.(7) The toxic byproducts of the bacteria are often present in sufficient amounts to permit anthrax toxin
detection in blood by immunoassay (analysis of antibody reaction to antigens), which can be performed in field laboratories.
(4)(5) If laboratory facilities are not available and specimens must be shipped, a sterile cotton swab can be soaked in blood
and air dried to promote sporulation. This can then be transported in a suitable sterile container to the nearest laboratory.[29]
A positive skin test to anthraxin (undefined antigen derived from the chemical breakdown of the bacillus and that was
developed and evaluated in the former Soviet Union) has also been reported to be of value in the retrospective diagnosis of
anthrax. Western countries have limited experience with this test. [301
Diagnostic methods for Infected Animals
Several laboratory methods for verifying anthrax in ill or dead animals may be employed, including smears, cultures, and
animal incubation. Direct microscopic examination of suspected material, when stained satisfactorily, will reveal
gram-positive bacilli that are 1 to 1.5 microns in diameter and 5 to 8 microns long. In blood smears, most of the bacilli will
be single cells, but short chains may exist if the animal has been dead for a few hours. Inoculating tryptose soy agar plates
with infected blood will show medusa-headed colonies in 12 to 24 hours. Postmortem diagnosis can be conducted on
7 of 30 11/6/2018 9:29 AM
Approved for Release: 2021/11/22 C06785544
Anthrax - 1ntellipedia
Approved for Release: 2021/11/22 C06785544
(b)(3) 50 USC 3605
Doc ID: 6640699
animals by collecting blood from any peripheral blood vessel. A blood smear should be prepared with gram or Giemsa stain,
and, if anthrax is the cause of death, large numbers of single to short-chained gram-positive, square-ended bacilli will likely
be present.P11[32]
Incubating inoculated guinea pigs or mice is extremely valuable. Ninety-five percent of the deaths resulting from animal
inoculation will occur on days 2, 3, and 4; the presence of B. anthracis should then be verified by smears taken from the
inoculated animals. Tissues may also be cultured for confirmation if an autopsy has been done; however, opening the
carcass, if anthrax is a likely diagnosis, is not advisable. Precautions against personal exposure must be taken.[331[34]
Properties/Persistence
B. anthracis is a gram-positive (stained cells remain bluish even when washed with organic solvents), encapsulated,
nonmotile (unable to move independently), aerobic (grows in the presence of air) bacterium. Its colonies are large, flat,
opaque, raised, and irregular. It produces rather large and stable spores. The spores are resistant to sunlight, heat, and
disinfectants. The optimal temperature for growth is 36 �C.
B. anthracis possesses three known virulence factors: an antiphagocytic capsule and two protein exotoxins, called the lethal
and the edema toxins. These toxins enable the bacteria to resist host defenses and invade host tissues via the bloodstream.
The survival period for the spores released in aerosol form is dependent upon release height, weather conditions, speed at
which the spores are released (deposition velocity), and height of temperature inversion. Spores in soil proves that spores are
more resistant to the sun's rays and other environmental elements when protected by loosely packed sand or dirt than
unprotected spores.
Ultraviolet (UV) light affects B. anthracis differently depending on whether the bacterium is in the vegetative state or spore
form. More than 99 percent of vegetative cells die after 20 seconds of exposure to UV rays; however, it takes 25 minutes of
UV exposure to kill the same amount of spores. Because B. anthracis spores are stable in the environment, this bacterium is
often considered the quintessential biological agent.
Properties of Anthrax
Bacillus anthracis is a gram-positive bacterium. The encapsulated cells appear in long, bamboo-shaped chains with square
or concave ends. The structure of the capsule is a high-molecular- weight polypeptide composed of poly-D-glutamic acid. It
is nonmotile (unable to move independently), and aerobic (grows in the presence of air). Its cells are large (5-10 microns
long and 1-1.2 microns wide), and colonies are flat, opaque, raised, and irregular, with a curled margin. B. anthracis grows
on ordinary blood agar within 18-24 hours. It produces spores in the center of the bacilli when in a medium or in the
environment rather than living tissues. These spores are ovoid, subterminal (band of colors on the end), stable, and do not
cause any significant swelling of the cells. The spores are also resistant to sunlight, heat, and disinfectants. The optimal
temperature for growth is 36 �C.[351
B. anthracis has three known virulence factors: an antiphagocytic capsule and two protein exotoxins (called the lethal and
edema toxins). The capsule is composed of a polymer of poly-D-glutamic acid, a compound that confers resistance to
phagocytosis. It may also contribute to the resistance of anthrax to lysis by serum cationic proteins. The genes encoding the
two protein exotoxins are located on a 60-kb plasmid. When bicarbonate, carbon dioxide and temperatures are at increased
levels, such as is found in the infected host, transcription of the genes for synthesis of the two toxins, as well as for the
capsule, are also increased. [36]
Like many bacterial and plant toxins, the anthrax toxins possess two protein components: a cell-binding, or B, domain and
an active, or A, domain that has the toxic and, usually, the enzymatic activity. The two toxins share the B protein, called
proteCtive antigen [molecular weight (MW) 83,000). The lethal toxin (lethal in experimental animals) is composed of the
protective antigen combined with the A protein, which is known as lethal factor (MW 90,000). The edema toxin, consisting
of the same protective antigen together with a third protein, edema factor (MW 89,000), causes edema when injected into
the skin of experimental animals. The three toxin proteins have no biological activity alone.1371
8 of 30 11/6/2018 9:29 AM
Approved for Release: 2021/11/22 C06785544
Anthrax - Intellipedia
Doc ID: 6640699
Persistence of Anthrax
Approved for Release: 2021/11/22 C06785544
(b)(3) 50 USC 3605
The persistence of spores that have been dispersed in aerosol is dependent upon the release height, weather conditions, speed
at which the spores are released (deposition velocity) and height of temperature inversion. The persistence of spores in soil
was testec The results showed that spores partially protected from the sun's rays
and other environmental elements by sand or dirt persist longer than unprotected spores. Also, the spores persist longer in
loose soil than in packed soil. The spores can survive in the soil for decades and can become airborne again if the surface is
disturbed; however, the inhalation hazard is reduced due to the large particle size.1381
Effect of Ultraviolet Radiation
B. anthracis is affected differently if it is in the spore form or the vegetative cell form. More than 99 percent of the
vegetative cells used in testing were killed within 20 seconds of exposure to UV wavelengths, while 25 minutes of UV
exposure were needed to kill the same amount of spores. In addition to being less sensitive to UV radiation, spores of B.
anthracis are much more resistant to drying and heat than vegetative cells. Because B. anthracis spores are stable in the
environment, anthrax is frequently considered the quintessential biological agent.139]
Mechanism of Exposure/Effects
B. anthracis begins its cycle of transmission in neutral or alkaline calcerous (chalky) soils that serve as an incubator for the
spores. Spores germinate to their vegetative form and multiply to infectious levels when favorable soil, moisture,
temperature, and nutrient conditions occur. The bacteria are found most commonly in areas that have periods of drought and
flooding. Flooding allows the bacteria to accumulate at the ground surface in low-lying areas. Subsequent drought affords
conditions for exposure of the spores. Epidemics tend to occur after heavy rainfall and flooding. Farm animals, especially
cows and sheep, become infected when feeding in these areas. People who tend to these animals or come into contact with
animal products are at high risk of acquiring anthrax. This is the natural way that anthrax is passed on to humans. Normally,
human anthrax is found in agricultural regions of the world.
Anthrax can cause infection in the human body through a number of routes. When it is contracted through abrasions on the
skin, it results in cutaneous anthrax. When it is contracted by the ingestion of contaminated meat, it results in intestinal
anthrax. When it is contracted by the inhalation of spores, it results in pulmonary, or inhalational, anthrax. The number of
particles that enter the bloodstream depends upon the particle size. Very few particles greater than 5 microns in diameter
penetrate past the nasal passages or the upper pharynx.
Mode of Transmission
Flies and other biting insects may transmit the disease; however, this is not the standard mode of transmission. Farm animals
may become infected by consuming contaminated feed such as bone meal, but they usually contract the disease by grazing
on contaminated vegetation. People who tend to these animals or come into contact with animal products are at high risk of
acquiring the disease. Dried or processed skins and hides of infected animals may carry spores for years. Human'anthrax is
normally found in agricultural regions of the world such as Southern Europe, Africa, Australia, Asia, North America, and
South America, where animals are frequently infected with anthrax.t40][411[42][431
Route of Infection
Bacillus anthracis can enter the human body through a number of routes, including abraded skin, contaminated food, and
inhaled spores. Each form of the disease is manifested differently according to its route of entry. Cutaneous (skin) anthrax is
the most common, naturally occurring form and accounts for nearly 90 percent of all cases. People who work with animal
products originating from anthrax-endemic areas are at risk of contracting the disease. Infection occurs from contact with
tissues of an animal or from the open wounds of an infected individual. Intestinal anthrax results from ingesting
contaminated meat. In industrialized nations, intestinal anthrax is less common than the cutaneous or pulmonary form of the
disease; however, intestinal anthrax is much more common than pulmonary anthrax worldwide. The low prevalence of
(b)(1)
9 of 30 11/6/20189:29 AM
Approved for Release: 2021/11/22 C06785544
Approved for Release: 2021/11/22 C06785544
Anthrax - Intellipedia
(b)(3) 50 USC 3605
(b)(1),
(b)(1)
(b)(1)
Doc ID: 6640699
intestinal anthrax in industrialized nations is presumably due to the stringent laws concerning animals allowed into the food
chain. Pulmonary anthrax is a form of the disease that is caused by inhaling spores from contaminated dust, wool, or hair,
especially when they are handled in a confined space. When inhaled, B. anthracis spores germinate in the alveoli. The
spores multiply and release three proteins: edema factor, lethal factor, and protective antigen. In specific combinations, these
proteins are potent toxins that enable the bacteria to resist host defenses and invade host tissues via the bloodstream. The
anthrax bacteria travel to the intestines and lymph nodes, where they can cause serious infection
Poisoning
The number of particles that enter the lungs depends upon the particle size. The smaller the particle, the farther it will
penetrate the respiratory system. Very few particles greater than 5 microns in diameter penetrate past the nasal passages or
the upper pharynx. The average size of the particle penetrating to the lungs is probably in the ranae of 1.0 to 3 5 microns
Toxicity
A respiratory dose of Bacillus anthracis is nearly always toxic, at least to humans.
There is no data as to how this relates to humans;
therefore, the monkey dose is the best estimate on the number of spores needed for a toxic dose of anthrax.[471[481[491
Prevention/Treatment
The best prevention against anthrax is vaccination. Bio Port produces the only licensed anthrax vaccine for human use. The
recommended schedule for vaccination is 0.5 nil injected under the skin at 0, 2, and 4 weeks, followed by boosters of 0.5 ml
at 6, 12, and 18 months. The vaccine induces an immune response in over 95 percent of humans who receive the initial three
doses.
A recommended treatment for those who contract any form of anthrax is to combine vaccination with the administration of
antibiotics. Animal studies have also demonstrated the effectiveness of supportive therapies, such as administering oxygen
and maintaining blood circulation. Treatment should begin as soon as exposure is suspected or anticipated for respiratory (in
the lungs) anthrax, which is the form of the disease most likely to be encountered in biological warfare and the most difficult
to diagnose
It is not clear it surviving an anthrax intection conters a lasting immunity tor
humans.
Animals appearing to be infected or having high temperatures should be given high dosages of antibiotics and be vaccinated
as soon as possible. Veterinary vaccines provide protective immunity starting 3 to 5 days after vaccination. Unlike vaccines
for humans, veterinary vaccines use live spores from the avirulent strains of anthrax.
Prevention
The best prevention against anthrax is vaccination. The currently available licensed vaccine is most effective against
cutaneous (on the skin) anthrax, the most common naturally occurring form of the disease. The vaccine is thought to be
effective against respiratory anthrax (based on animal studies), the form of the disease most likely to be encountered in
biological warfare. Vaccination is recommended for high-risk populations, such as people in direct contact with anthrax-
infected animals or animal products (i.e., farmers, factory workers, and laboratory or medical personnel) and soldiers who
are at risk of an anthrax attack. Approximately 150,000 service members were vaccinated between 11 January and 28
(b)(1)
10 of 30 11/6/2018 9:29 AM
Approved for Release: 2021/11/22 C06785544
Anthrax - Intellipedia
Doc ID: 6640699
Approved for Release: 2021/11/22 C06785544
(b)(3) 50 USC 3605
February 1991 (25 to 30 percent of the total US forces deployed during the Persian Gulf War). On 3 March 1998, Secretary
of Defense William Cohen announced that all U.S. military personnel deployed to the Gulf region would be vaccinated
against anthrax. Eventually, all active duty and reserve personnel will be vaccinated.[50[511[52]
Vaccines
Bio Ports produces the only licensed human vaccine against anthrax. The purpose of a vaccine is to stimulate an immune
response in the body to establish resistance to the disease. With this vaccine, the body's immune response is stimulated by
the presence of protective antigen material, a component of the toxins produced by the Bacillus anthracis bacteria. The
protective antigen vaccine induces an immune response in over 95 percent of humans who receive the initial three doses.
The vaccine should be stored at refrigerator temperature (between 2 and 8 �C) and not frozen.[53][541[551[56]
In the US, the protective antigen vaccine, based on research since the mid-1940s, has replaced the weakened live dose type
of vaccine developed by Pasteur. As research continues, there is speculation that the current vaccine will probably be
replaced by products of recombinant DNA research developed by the US Army Medical Research Institute of Infectious
Diseases; however, there are many arguments against recombinant vaccines. In the former USSR, a weakened live dose is
still used for human vaccination. Its developers claim it is reasonably well tolerated and affords some protection against
cutaneous anthrax in clinical field trials.[571[58][591
Inoculation Schedule and Resulting Protection
The recommended schedule for vaccination is 0.5 ml injected under the skin at 0, 2, and 4 weeks, followed by boosters of
0.5 ml at 6, 12, and 18 months. Annual boosters are recommended if the potential for exposure continues. Limited human
data suggest that completing only the first three doses of the recommended six-dose primary series can provide good
protection against both cutaneous and respiratory anthrax. Studies in rhesus monkeys indicate that good protection against
respiratory anthrax is achieved after two doses (1 to 16 days apart) for up to two years. It is likely that two doses in humans
would be protective as well, but there is too little information to draw firm conclusions. As with all vaccines, the degree of
protection depends upon the magnitude of the exposure; a large dose of the infectious agent could overwhelm the vaccine
protection.[601[61]
Recent Russian research has suggested that an altered form of some strains of Bacillus anthracis (B. anthracis) may
overcome vaccine protection. Four scientists from a government research center experimented with adding genetic factors to
B. anthracis. Their results, published in mid-I997, claimed that adding the ability to destroy red blood cells (hemolysis) to
B. anthracis allowed the altered strains to defeat immunity (in animals). The scientists also claimed that an altered vaccine
could easily be created to defeat such altered strain5.[621
Sensitivities and Side Effects
Vaccination is not appropriate for anyone who is sensitive to any of the vaccine components (i.e., formalin, alum,
benzthonium chloride) or has a history of clinical anthrax. For those who are vaccinated the likelihood of side effects is low.
Up to six percent of recipients will experience mild discomfort, including tenderness, redness, swelling, and itching, at the
inoculation site for up to 72 hours. These reactions peak at one to two days and usually disappear within two to three days.
Less than one percent will experience more severe reactions in the general area (arm or leg) of the inoculation, potentially
limiting the use of the extremity for one to two days. Modest systemic reactions (i.e., muscle pain, malaise, low-grade fever,
and headache) are uncommon and usually last for one to two days. Severe systemic reactions (i.e., Anaphylaxis, an extreme
allergic reaction, that precludes additional vaccination) are rare. There are no long-term conditions resulting from local or
systemic reactions.[63][64]
Treatment
The most effective treatment for all forms of anthrax is to combine vaccination with the immediate (before symptoms appear
if possible) administration of antibiotics. The use of antibiotics keeps the patient alive until the patient's body can build
immunity to anthrax via vaccination. Studies on rhesus monkeys have shown that the concurrent use of more than one
antibiotic has a synergistic effect (interaction of two or more is greater than sum of individual elements), which increases the
11 of 30 11/6/20189:29 AM
Approved for Release: 2021/11/22 C06785544_
Anthrax - Intellipedia
Approved for Release: 2021/11/22 C06785544
(b)(3) 50 USC 3605
Doc ID: 6640699
patient's chance for survival. This can also help treat the antibotic-resistant strains of anthrax that have been developed.
[65][66][67][681
Animal studies have also demonstrated the effectiveness of supportive therapies, such as administering oxygen and
maintaining blood circulation (e.g., by using isoproterenal). Oxygen administration helps to overcome the decrease in blood
oxygen levels caused by anthrax toxins. Using a positive-pressure respirator helped anthrax-infected rhesus monkeys
survive. Isoproterenol facilitates circulation and oxygenation of the blood by increasing cardiac output and improving blood
circulation in the lungs. Rhesus monkeys given isoproterenol survived the effects of anthrax toxin.[69]
Treatment should begin as soon as exposure is suspected or anticipated for respiratory anthrax. If antibiotic therapy is not
begun until after symptoms appear, death is a likely result. The antibiotics may be administered orally or intravenously,
depending on the form and severity of the disease. The duration of the administration also depends on the form of the
disease.M[711172]
Under Battlefield Conditions
(b)(1)
If clinical signs of anthrax occur, patients should continue to receive antibiotic treatment.[731[7411751
In the event of an anthrax attack, inoculations should begin immediately for those soldiers who did not complete an
immunization program. A single 0.5- mL dose of vaccine should be given under the skin, followed by two 0.5-mL doses of
the vaccine given two weeks apart. Those previously vaccinated with fewer than three doses should receive a single 0.5-mL
booster, and vaccination probably is not necessary for those who have received the entire three-dose primary series. If the
vaccine is not available, antibiotics should be continued beyond four weeks while the patient is closely observed to confirm
that anthrax is not present.[76]
For Respiratory Anthrax
Respiratory anthrax should be treated with large doses of intravenous antibiotics (two million units administered every two
hours). Penicillin, ciprofloxacin, and doxycycline are the preferred drugs. Penicillin is probably the best known and most
widely used. Ciprofloxacin and doxycycline are recommended to overcome penicillin-resistant organisms. Tetracycline,
erythromycin, and chloramphenicol have also been used successfully. Laboratory tests suggest that gentamicin, cefazolin,
cephalothin, vancomycin, clindamycin, and imipenem could also be used successfully to treat anthrax. Streptomycin and
penicillin administered together have had synergistic effects on respiratory anthrax disease in rhesus monkeys. Such efficacy
remains to be demonstrated in humans.[77][781
For Cutaneous, Intestinal, and Oropharyngeal Anthrax
Cutaneous anthrax that is localized and does not appear to be spreading may be treated with oral penicillin. If the infection
begins to spread or systemic symptoms are present, then intravenous therapy with high-dose penicillin (two million units
administered every two hours) should be initiated. Treatment should be continued for 7 to 10 days. This therapy, if effective,
will reduce systemic symptoms and swelling. The treatment will not change the evolution of injured skin tissue, which will
progress through ulceration, sloughing, and scar formation. Intestinal and oropharyngeal (in the mouth) anthrax should be
treated with large doses of intravenous penicillin (two million units administered every two hours).[79][801
Immunity
It is not clear if surviving an anthrax infection confers a lasting immunity for humans. Hodgson reports two cases of
cutaneous anthrax that seemed to result in limited immunity. One case occurred in a veterinary surgeon who had an infected
forearm from which Bacillus anihracis was isolated. This patient had anthrax three years previously that had been
bacteriologically verified and that had healed with scar formation. The second attack was accompanied by minor swelling
and was readily cleared by two injections of serum. The second case reported by Hodgson was a teak buyer who developed
a non-inflamed blister on the back of his neck. A brownish fluid obtained from the blister contained unidentified bacilli. The
12 of30 11/6/2018 9:29 AM
Approved for Release: 2021/11/22 C06785544
Anthrax - Intellipedia
Approved for Release: 2021/11/22 C06785544
(b)(3) 50 USC 3605
Doc ID: 6640699
teak buyer apparently had anthrax in severe form five months previously, that had been treated with serum and surgical
excision. A second attack, six weeks after the first, was less severe and, when treated in the same fashion as the first, quickly
subsided.[811
For Exposed Animals
All exposed animals, whether or not they appear to be infected or have high temperatures, should be isolated and given
antibiotics and vaccinations. Infected and exposed animals should be in a separate location from non-exposed animals and
given long-acting antibiotics immediately. High dosages of antibiotics (50 mL of long-acting penicillin; 300,000 IU/mL) are
recommended. Such antibiotic therapy can stop anthrax intoxication if given early. Infected or exposed cattle should be
vaccinated as soon as possible using Thraxol (Miles Laboratories) or Anthrax Spore Vaccine (Colorado Serum). The vaccine
provides protective immunity starting 3 to 5 days after vaccination. A booster vaccination should be given according to label
directions. Antibiotic therapy can prevent death until the vaccine can provide immunity. [82]
Veterinary vaccines use live spores of the non-disease-producing strains of anthrax. Antianthrax serum of bovine and equine
origin is marketed in 100 mL quantities by the Pitman-Moore Company of Indianapolis, Indiana. Preliminary precautions
against serum sensitivity must be observed. [83] [84]
Alternate Uses
There are no developed alternative uses for B. anthracis; however, a vaccine that uses a potent toxin from B. anthracis is
being studied by researchers at Harvard Medical School. This vaccine could lead to an entirely new class of human vaccines
against most viruses, certain bacteria, and parasites. This development is the first successful attempt to engineer a
protein-based vaccine that works by using the immune system's killer T cells. T cells respond to infection and generate a
specific immunological memory for future protection. Researchers have designed the vaccine by fusing a harmless piece of
an anthrax toxin with a piece of the model pathogen to stimulate the T cells. The transporter component of the anthrax toxin
is then added to this mixture and injected into mice. Although the initial work appears promising, it is uncertain whether the
vaccine can protect against death by anthrax. Steps still need to be taken before the vaccine is tested on humans.
Vaccine
Although there are no alternative uses for Bacillus anthracis (B. anthracis), researchers at Harvard Medical School are
building an experimental vaccine from a toxin of B. anthracis. This vaccine is being used to combat a model pathogen and
could lead to an entirely new class of human vaccines against most viruses, certain bacteria, and parasites. Also, it may be
helpful in developing cancer vaccines and therapies.[851
Immune system
The work being done at Harvard represents the first successful attempt to engineer a protein-based vaccine that works by
using the body's natural immune system killer T cells. T cells respond to infection and generate a specific immunological
memory for future protection. Most current protein-based vaccines, such as the one used against tetanus, stimulate B cells. B
cells produce antibodies and only detect invading pathogens so long as they are outside of cells. Once the pathogen invades
past this point of defense and slips inside cells of the body, T cells must be activated. This requires that the antigens they
combat be displayed to them from inside infected cells. Delivering a vaccine into cells is much more complex than simply
injecting it into a person's bloodstream.1-861
Inserting the vaccine
Researchers have engineered an intracellular vaccine that moves the proteins across the cell membrane and into the
cytoplasm. A technique has been developed to manipulate some of the toxin's components so that they are innocuous but
able to transport any protein. Next, the researchers genetically fuse a harmless piece of the anthrax toxin to a piece of their
model pathogen required to stimulate T cells but unable to cause full-blown disease. Finally, they mix the pieces with a
transporter component of the anthrax toxin and inject it into mice.[87-I
13 of 30 11/6/20189:29 AM
Approved for Release: 2021/11/22 C06785544
Anthrax - Intellipedia
Doc ID: 6640699
Results of the vaccine
Approved for Release: 2021/11/22 C06785544
(b)(3) 50 USC 3605
Although the initial work appears promising, steps need to be taken before the vaccine is tested on humans. AIDS patients
and initial testing of vaccine information removed; could be misleading.[88]
Production/Storage
In order to produce any type of bacterial agent, including B. anthracis, seed stocks were
carefully selected to provide the desirable characteristics.
production wasia relatively simple process. B. anthracis grew readily on a wide variety
of substrates; however, sporulation could vary widely depending on conditions and
media used. The media were stored for several days before they were inoculated with B.
anthracis, ensuring that the culture medium was sterile and not contaminated with other
bacteria.
(LIA0U0)
Materials Indicatiita B. anthracis
Culture and Wenoonizatiiin4
R. anthracis culture or reed stock
Lab Glassware and Supplies
Groirth Media
Centrifuge
Incubator
pH Meta-
Refrigerator
Thallium Acetate"
Freeze The,
Microscope
Shaker
.Fermeztter
Milling Equipment
Fluidizer or Silica
A hazard of B. anthracis was the containment of the spores after production. If released into the air, the bacteria could enter
either the air-handling system or environment where the persistent spores could contaminate people and the surrounding
area.
Pilot Scale Production
Interim Report 108
Experimental Operating Procedure for Manufacturing B anthracis in Pilot Plant B-4,
(C) As a reference for comparative analysis of state biological warfare programs, this document serves as an excellent
guidepost. Technologies presented in this tome are dated, but demonstrate how the US was able to arrive at ton production
capabilities for anthrax BW agent using extant and newly developed or improvised technologies for BW production. The
document has been declassified to Confidential.
Del 20:43, 28 November 2007 (UTC)
BW Production Requirements in the US BW Program (C)
(C) Special report 243 provides unique insights into the challenges posed in 1956 by modifications of an existing production
line to adapt to a new biological warfare agent, anthrax. For analysts involved in evaluation of production processes for
(b)(1)
(b)(1)
(b)(1)
14 of 30
Approved for Release: 2021/11/22 C06785544
11/6/20189:29 AM
Anthrax - Intellipedia
Doc ID: 6640699
Approved for Release: 2021/11/22 C06785544
(b)(3) 50 USC 3605
biological warfare, the document give real-world examples of how the US modified a processes line to make anthrax
biological warfare agent.
Since this facility
was aesignea specincaily tor proaucuon ana processing or vegetative agents, extensive moaincation are required to prepare
the facility for production of N. Major revisions are required in agent concentration, filling, loading, clustering and waste
sterilization areas. These revisions include: installation of vessels for collecting and blending concentrated suspensions of
agent from the centrifuges; addition of equipment for filling, loading and clustering munitions; and addition of equipment
for decontaminating agent in liauid effluent from potentially contaminated areas Minor revisions are reatiired in all nmcess
areas.
This document is in 2 parts. Part 1 ] Part 2 ] Del 19:27, 16 November 2007 (UTC)
Laboratory requirements
In order to produce liquid cultures of Bacillus anthracis, a well-equipped microbiologgy laboratory, including the following
laboratory equipment and supplies is required:
� Mettler balance, model PM600
� Beakers, graduated (100 ml and 600 ml)
� Erlenmeyer flasks, polycarbonate with screw cap (500 ml and 1000 ml)
� Magnetic stirrers
� Magnetic stirring bars
� Microscope
� Glass microscope slides
II Autoclave, portable. electric
� Mini-pH meter
� Pipets, disposable, sterile (5 ml and 10 ml)
E Pipet filler
� Shaker, rotating
� Shaker, flask carrier for 15-500 ml flasks
� Digital thermometers
� Medium components
� Tryptic soy broth
� Peptone
� Glucose
� Plasmolyzed yeast
� K 2HP0 43(H 20)
� KH 2P0 4
� FeS0 47(H 20)
� MnS0 44(H 20)
� MgS0 47H 20
� CaC126(H 20)[891
Seed selection
In order to produce a bacterial agent, including B. anthracis, seed stocks are carefully selected to provide the desirable
characteristics (e.g., stability, virulence, uniformity, good growth and yields, and aerosol stability).
After testing for purity and culture
characteristics, the seeds are maintained at a liquid nitrogen temperature (195.8 �C) until needed.901
(b)(1)
(b)(1)
(b)(1)
15 of 30 11/6/2018 9:29 AM
Approved for Release: 2021/11/22 C06785544
Anthrax - Intellipedia
Doc ID: 6640699
Approved for Release: 2021/11/22 C06785544
(b)(3) 50 USC 3605
Starter cultures
To start the culture production process, seed stocks are reconstituted with saline solution. The seed stocks must be able to
withstand a series of at least five serial transfers for build-up to the desired volume. To create the desired volume, the seed
cultures are prepared by inoculating 500 ml of media with stock cultures and incubating them at 35 �C. Fifty milliters of the
stock culture are used to inoculate 500 ml of media. In order to achieve maximum growth and induce spore formation, the
cultures are aerated by shaking on a rotating shaker. Once the cultures have achieved maximum growth, as measured by a
spectrophotometric evaluation of the turbidity (cloudiness) of the cultures, the culture flasks can be stored in a refrigerator
until the desired amount of culture has been obtained.[911
Media production
Culturing B. anthracis is a relatively simple process. It grows readily on a wide variety of substrates (material on which an
organism lives); however, sporulation can vary widely depending on conditions and media used.
Distilled
N,vater snoula oe usea to prepare meaia. 1 lie medium without glucose is prepared and distributed in 20-ml amounts into
500-ml Erlenmeyer flasks. The flasks are stoppered with cotton plugs and then covered with aluminum foil. The flasks are
sterilized at 20 pounds pressure for 20 minutes in an electric autoclave or pressure cooker with a pressure gauge. The
glucose is sterilized separately in distilled H 20. The glucose is added to the medium after cooling. The culture medium is
stored for several days before inoculation to ensure that the culture medium is sterile and not contaminated with other
bacteria.1921[93]
In the media preparation for industrial size cultivation, the dry or wet media ingredients are added to treated water. The
sterile water is contained in an agitated stainless steel, jacketed vessel to allow temperature control. Steam, hot, and cold
water can be circulated in the jackets to speed dissolution of the ingredients, as well as hold the media for subsequent
sterilization. Generally, sterilization of prepared media is performed in another separate, closed, agitated stainless steel
vessel while the fennentor and associated piping is being separately sterilized by high-pressure steam. Following
sterilization of the media, the media is transferred under aseptic conditions to the fermentator.
Technical report BWL 12
Evaluation of Media for Growth of Bacillus anthracis and Pasture/la tularensis, Apr 1959
(C) This ['document ] focuses on the selection and optimization of nutrient media for industrial-scale production of anthrax
and tularemia biological warfare agents. The detailed information and references in this document are invaluable resources
(b)(1)
(b)(1)
'6 of 30 11/6/20 I 8 9:29 AM
Approved for Release: 2021/11/22 C06785544
Anthrax - Intellipedia
Approved for Release: 2021/11/22 C06785544
(b)(3) 50 USC 3605
Doc ID: 6640699
for technical evaluations of biological warfare capabilities.
Del 21:26, 28 November 2007 (UTC)
Fermentation
An agitated stainless steel vessel can be used to cultivate B. anthracis. Temperature, dissolved oxygen, and pH can be
controlled in the fermentor. Production of B. anthracis can be accomplished in 5,000-gallon, closed, aerated vessels.
Associated piping should also be stainless steel, completely welded, and designed for high-pressure steam sterilization. All
valves should be Saunders diaphragm types, modified to allow steam to pass through all areas of the system to ensure
complete sterilization. Air for the vessels must be passed through filters to ensure purity and reduce contamination of the
cultures.
Centrifugation
(b)(1)
Hazards of production and storage
A hazard of B. anthracis production is the containment of the spores after production. If released into the air, the bacteria
can enter the air-handling system and the environment where the spores can contaminate the surrounding area. At present,
(b)(1)
Detection
Although current technology cannot provide immediate identification of biological agents, detectors can distinguish between
some biological warfare (BW) agents and natural organic matter present in the atmosphere in near real time (seconds to
minutes). Also, positive identification of an agent is possible within minutes using identifiers. Because detection devices do
not provide immediate recognition of BW agents, detection systems must be used in conjunction with other measures such
as medical protection (vaccines and other prophylactic measures), intelligence, and physical protection to provide layered
primary defenses against a biological attack.
Role of detection
Because B. anthracis aerosols have similar compositions and external characteristics and behave as natural, organic matter
present in the atmosphere, detection of B. anthracis can be difficult.
(b)(1)
One of the greatest benefits of biological detection is the ability to adjust plans and courses of action to the situation at hand.
The following could be a typical scenario if a B. anthracis attack occurred. First, intelligence conveys information
suggesting a possible anthrax attack. Second, personnel employ detection systems to confirm the presence of the agent.
Simultaneously, physical and protective measures are implemented to protect the population from the onset of disease until
positive identification has been confirmed. A typical physical measure is the protective mask. It is worn to prevent potential
17of30 11/6/2018 9:29 AM
Approved for Release: 2021/11/22 C06785544
Anthrax - Intellipedia
Approved for Release: 2021/11/22 C06785544
(b)(3) 50 USC 3605
Doc ID: 6640699
B. anthracis particles from reaching the lungs. Finally, medical personnel assist the exposed population and vaccinate
everyone. A primary concern involving a B. anthracis attack is that respiratory anthrax has a mortality rate approaching 100
percent unless treatment is begun before signs and symptoms of the disease appear. If anthrax is detected before symptoms
appear, the chances of survival are increased because vaccines and other therapeutic measures can be administered. Interim
systems of detecting biological agents are being fielded in limited numbers. Without these measures, the first indication of
an attack could be the unprotected, ill soldier.[971[98]
Architectural hardware
Since 1992 and Operation Desert Storm, there has been increased interest in the development of biological agent detection
systems. Most bio-detection systems currently in use or under development have several functional components. These
components include a trigger, a collector, a detector, and an identifier. Since actual biological warfare (BW) attacks vary, the
detection equipment used depends upon the situation at hand. Therefore, it is important to understand the specific functions
for each piece of equipment. The following paragraphs describe the function of each device, and the tables at the end of this
article give examples of the many components availableJ991
Triggers
The trigger's function is to provide an early warning that a change in background air has occurred. Unless a trigger is used
before and during an actual airborne release of the anthrax agent, it will not offer a significant response since it can only
verify a change in the air. The indicators used to identify a change in the background levels are an increase in the aerosol
particle count and an increase in the fluorescence of biological-type aerosols.[991
Collectors
A collector or concentrator samples the atmosphere and concentrates the airborne particles into a liquid medium for analysis.
A collector is beneficial to use when an anthrax attack occurs, because the artificial particles are present in the air. When a
collector is used with a trigger, the trigger relays a signal to the collector indicating a change in the background level. As
soon as the collector receives the signal, an air sample is collected and airborne particles are concentrated into a liquid
medium. This scenario would take place only in a situation where continuous monitoring is occurring.[991
Detectors
Detectors indicate the presence of biological matter in a collected sample. They are non-specific in their detection method;
therefore, they respond to both pathogens (something that causes disease) and non-pathogens. Because detectors are
non-specific to the biological substance they detect, specific identifiers must be used to confirm the presence of a BW agent.
In a usual scenario, a trigger responds to a change, a sample is collected, a detector then senses biological materials, and an
identifier confirms the presence of a specific BW agent, such as anthrax. (see Table 3).
Detectors indicate the presence of biological matter in a collected sample. They are non-specific in their detection method;
therefore, they respond to both pathogens (something that causes disease) and non-pathogens. Because detectors are
non-specific to the biological substance they detect, specific identifiers must be used to confirm the presence of a BW agent.
In a usual scenario, a trigger responds to a change, a sample is collected, a detector then senses biological materials, and an
identifier confirms the presence of a specific BW agent, such as anthrax.1991
Identifiers
The role of identifiers is to analyze a collected liquid sample to determine whether a particular BW agent, such as anthrax, is
present in the sample. Identifiers are typically based on antigen-antibody responses. There are three methods of feeding a
sample to an identifier. The first method is pumping the sample directly into the identifier from the collector. The second
method is using a syringe to inject the sample into the identifier's inlet line. The sample is analyzed for the presence of
anthrax. This is usually done in laboratory settings. The final method feeds a sample to an identifier by placing droplets of
the collected sample on "Tickets." This mainly requires taking a field sample using a swipe kit.[991
18 of30 11/6/2018 9:29 AM
Approved for Release: 2021/11/22 C06785544
Anthrax - Intellipedia
Doc ID: 6640699
Point detectors
Approved for Release: 2021/11/22 C06785544
(b)(3) 50 USC 3605
One method of biological warfare (BW) agent detection is use of the point detector. An example of a point detector is an air
sampler. Air samplers, collect samples of viable particles in multiple stages.
Each stage contains a number ot precision-drilled orifices; they are a constant size for each stage. The size of the orifice is
based on the particle size desired to be collected in that stage. Orifice sizes decrease with each succeeding impactor stage.
Particle-laden air enters the instrument (drawn into the impactor using a blower or pump of some type), and the airborne
particles are directed towards the collection surface by the jet orifices. Any particle not collected by that stage follows the
stream of air around the edge of the collection surface to the next stage. The collection plate is typically a petri dish with
agar (growth media). A selective agar specific to the organism or an all-purpose bacteriological medium can be used. After
sampling, the collection plates are removed, covered, and placed in an incubator. After incubation, the number of colonies
on each plate is counted using standard bacteriological counting techniques. These samples must be given to another device
for anthrax detection (i.e., to identifiers), because air samplers cannot detect agents, they only sample air.1I001
Mobile Labs
Another method of detection is the use of mobile laboratories, such as the
tests environmental air
samples by concentrating appropriate aerosol particle sizes in the air, then subjecting the sample to antibody-based detection
for selected agents. An improved version is under development to upgrade and expand point detection capabilities.[101][102]
Another example of a mobile laboratory was designed during
that identified potential biological warfare (BW) agents in air
SCLIIIIJICS. /AIM], tii yswin U11 CI I pill, WHOM 1,,G11 immunosorbent assay (ELISA) which quickly identified
selected bacterial agents. The ELISA immunoassay test depends on the detection of a highly specific reaction (binding) of
antigens with their corresponding antibodies (antigen-antibody complex). In an immunoassay-based BW agent identification
system, the presence of an agent is detected and identified by relying on the specificity of the antigen-antibody binding
agent. In the Persian Gulf, early research concentrated on the identification of Bacillus anthracis whole cells, and the
resulting assays were fielded in the Persian Gulf. Anthrax could be reliably detected in 5.5 hours. An assay with shortened
incubation times was later developed (assay run time of 3.0 to 3.5 hours).[I�31[104]
Stand-off detectors
The third method of detection is stand-off detectors. Two examples are the
s in the research and development phase.
Both systems use
Remote detection of biological warfare (BW) agents is best accomplished using
Both methods of detection
will provide early warning, avoid contamination, and point out other detection assets.1I061
(b)(1)
(b)(1)
(b)(1)
(b)(1)
(b)(1)
(b)(1)
19 of 30 11/6/20189:29 AM
Approved for Release: 2021/11/22 C06785544
Anthrax - Intellipedia Approved for Release: 2021/11/22 C06785544
Doc ID: 6640699
Anthrax as a Biological Warfare Agent
US CBW Program Data
(b)(3) 50 USC 3605
(U/FOUO) This 1972 compilation of data derived from US BW program activity is unique and not available elsewhere. It is
a valuable analytical tool, This is one of a set of Volumes created for the purpose of retaining the unique information that
was derived from actual weapons research. This Secret document is exempt from declassification.
(U) [IPart One of Volume VII of the Joint CB Technical Data Source Book part I] [I Part 2 ]presents the characteristics of
the bacterium, Bacillus Ant hracis, that relate to the possible use of the organism as an agent in biological warfare and to
defense against such use. Parameter values are presented for B. anthracis that may be used with the general models for
predicting weapons effects presented in Volume X. In addition, where adequate information vas available; models and
parameter values unique to B. anthracis are presented in this volume. These weapons effects estimates may then be used to
assess defensive requirements. Del 15:32, 16 August 2007 (UTC)
Weaponization/Dispersal
Anthrax is considered the prototypical biological warfare (BW) agent. Its spore-forming ability makes anthrax well suited
for delivery by missiles or bombs. The weapons that were used to carry anthrax were
Jelivery system. These weapons were designed to be loaded into a rocket or
missile warhead.
Each type of weapon was capable of housing either a liquid or dry agent fill. Anthrax could have been
employed as a strategic, tactical, or covert weapon. In its strategic capability, anthrax could have been used for preemptive
attacks, but only if a country had the appropriate delivery systems. In its tactical capacity, anthrax could have been used for
attacks on reserve formations and service support organizations. In its covert capacity, anthrax could have been used to
attack agricultural systems or as an anti-animal weapon.
Weapon Types
Anthrax is considered to be the prototypical biological warfare agent. Its spore-forming ability makes it well suited for
delivery by missiles or bombs. Anthrax can be used as a strategic, tactical, and covert weapon. Each type of weapon is
described in the following paragraphs.
As a strategic weapon, anthrax could be Used for preemptive or initiating attacks, but only if the attacking country has
appropriate delivery systems and has taken into consideration the time (three to five days) that the infection takes to cause
mortality.
to disseminate anthrax,
either overtly, or covertly. In the strategic role, an anthrax warhead could be used as a wide-area-effect weapon that causes a
country's medical system to be overwhelmed and closes airfields/airports or seaports.
As a tactical weapon, anthrax could be used for attacks on reserve formations and service support organizations.
As a covert weapon, anthrax could be used as an anti-animal weapon. It could also be used as an assassination weapon;
however, because anthrax is slow acting, it may not be effective. Anthrax could also be used as an antipersonnel weapon
Bomblets
(b)(1)
(b)(1)
20 of 30
Approved for Release: 2021/11/22 C06785544
11/6/20189:29 AM
Anthrax - Intellipedia
Doc ID: 6640699
Approved for Release: 2021/11/22 C06785544
(b)(3) 50 USC 3605
Types of Bomblets
Line Source Weapons
Other Delivery Systems
A wide range of potential delivery systems was studied in the United States prior to 1969. It was found that these systems
examined the release of dry agent as well as wet releases
Meteorological Conditions for Release
Meteorological conditions necessitate precise planning of a biological warfare (BW) attack. Bacillus anthracis is refractory
(resistant) to sunlight. Wind is also an important factor in preplanning a BW attack.
(b)(1)
(b)(1)
(b)(1)
(b)(1)
(b)(1)
21 of 30 11/6/20189:29 AM
Approved for Release: 2021/11/22 C06785544
Anthrax - Intel' ipedia
Doc ID: 6640699
Approved for Release: 2021/11/22 C06785544
(b)(3) 50 USC 3605
When anthrax is used with an
When discussing aerosols, it is important to discuss the level at which primary infection is initiated or the degree of
penetration into the trachea-bronchial tree. To a great extent, the size and mass of the inhaled particle determine initiation
and penetration.
very few particles greater than five microns in diameter penetrate past the nasal
passages.[I19]
Dispersal Efficiency
The dispersal efficiency for anthrax depends on the type of weapon or device used to disseminate the agent and whether the
agent is in liquid or dry form. The following chart compares the efficiency of liquid and dry anthrax dissemination from
bomblet and line-source weapons. [120]
The means by which a biological warfare (BW) agent is delivered on target is associated with the munition;
frequently, the munition dictates the delivery system.
theWith evolution of sophisticated line-source hardware, the agent, munition, and delivery system must be
carefully integrated.[I21]
Meteorological Conditions for Dispersal
The meteorological conditions required for anthrax dissemination necessitate precise planning of a biological warfare (BW)
attack. Most BW attacks, regardless of agent, are likely to occur shortly before daybreak, at sunset, or at night. Wind is an
important factor in preplanning a BW attack.
I hos, the ettects on the target
cannot be predicted. Liquids and dry agents can be disseminated effectively over a wide range of environments--
sub-freezing (for dry agent) temperatures. hot tropical conditions dry-desert conditions and during moderate rain and
snowfall. As a general rule
Physical Protection/Decontamination
Physical protection from biological hazards requires the use of respiratory protective equipment. One of the most important
items of personal protective equipment is the protective mask and associated filters. This type of protection dramatically
reduces inhalation exposure to anthrax spores. The protective value of the mask is critically dependent on the fit.
Protective clothing provides a barrier between a soldier and potentially hazardous agents. The various levels of protective
clothing are dependent upon the anticipated presence of the agent.
Decontamination is the process by which an agent is removed or the agent concentration is decreased so that it no longer
poses a hazard. When anthrax spores contaminate the soil and vegetation via aerosol dispersal, decontamination of those
(b)(1)
(b)(1)
(b)(1)
22 of 30 11/6/2018 9:29 AM
Approved for Release: 2021/11/22 C06785544
Anthrax - Intellipedia
Doc ID: 6640699
Approved for Release: 2021/11/22 C06785544
(b)(3) 50 USC 3605
areas is nearly impossible. Methods of decontamination for bacterial agents include burning the area or spraying it with a
mixture of bleach and water. Chemicals used for decontamination of other biological agents may be used to decontaminate
areas exposed to anthrax. Absolute proof of soil and vegetation decontamination is difficult, because anthrax is a naturally
occurring organism. Protective Mask
Using a protective mask dramatically reduces potential exposure to inhaled Bacillus anihracis spores.
The protective value of the mask is critically dependent on the fit. If (b)(1)
fitted improperly, the protective ratio afforded by the mask has been measured to be as low as 10, which would not provide
an effective barrier to B. anthracis spores.[123]
The respirator mask is designed for use with a specific type of filter. Some masks are designed to be worn with two
filters.[124]
Aerosol delivery systems for biological warfare agents most commonly generate invisible clouds with particles or droplets
of less than ten micrometers. They can remain suspended for extended periods. Particles may adhere to individuals, their
clothing, or their masksJi 251
Protective Clothing
There are various levels of protective clothing. The level of clothing required is dependent on the anticipated magnitude of
agent exposure. Protective clothing prevents chemical and biological agents from penetrating through layers of clothing and
contacting the skin. Permeable protective clothing allows air and moisture to pass through the fabric without hindering the
protection capabilities of the garment. The overgarment, undergarment, gloves, and boots are considered protective
clothing.[126]
Decontamination Exposure
In a biological attack, anthrax would likely be dispersed as an aerosol, exposing large areas of soil and vegetation to anthrax
spores. Decontamination of areas exposed to anthrax is nearly impossible. It takes decades of weathering and human
intervention to properly decontaminate an area exposed to high doses of anthrax. In infected livestock, the bacteria might
create new reservoirs for the disease, making the affected area impossible to use until the area is totally decontaminated.
Animals dying from anthrax often bleed from their body orifices prior to death, thereby contaminating soil or bedding.
Animal carcasses infected with anthrax should be incinerated at the site of death.[127][128][129]
Methods of Decontamination
Decontamination methods for all types of bacterial agents include burning the area or spraying the area with a mixture of
bleach and water [seven parts Supertropical Bleach (STB) and 93 parts water]. Spraying water or oil on the area helps
prevent secondary aerosol exposure, but does not decontaminate the bacteria. Anthrax spores are highly resistant to
decontamination.[IA
Chemicals Used for Decontamination
Any commercial hypochlorite (bleach) can be used to produce a decontaminant that will rapidly kill all potential biological
threat agents, including Bacillus anthracis spores. Chlorine dosages sufficient to kill anthrax spores rapidly would kill other
microorganisms even faster. Sodium hypochlorite, formaldehyde, and phenol are also effective sporicidal decontaminants.
These chemicals are caustic and corrosive in addition to being toxic and offensive to humans and animals. A new
commercial sporicidal product, Exspor, has been found to be less corrosive than hypochlorite bleach, not caustic, and
generallyharmless to humans; however, inhalation of the aerosolized vapors during decontamination may result in breathing
difficulties due to the acidity of the solution.[1311[132]
Chemicals used for the decontamination of other biological agents are also of some use for anthrax exposures. These
chemicals include:
23 of 30 1 1/6/201 8 9:29 AM
Approved for Release: 2021/11/22 C06785544
Anthrax - Intellipedia
Approved for Release: 2021/11/22 C06785544
(b)(3) 50 USC 3605
Doc ID: 6640699
� 10 percent sodium hydroxide (caustic soda or lye) or potassium hydroxide (caustic potash) decontaminant combined
with water (highly corrosive and toxic)
� Calcium hypochloride: faster acting than SIB; only use if SIB is not available; can be used in dry or slurry form
� Formalin (formaldehyde): recommended as decontaminant for relatively closed areas
� Peracetic acid: used on equipment and utensils
� Ethylene oxide: used in areas that can be made airtight
� Carboxide: consists of a mixture of ethylene oxide and carbon dioxide; airtight enclosure required
� Sodium hypochlorite solution (household bleach): used for cutaneous exposure diluting two parts bleach to 10 parts
water
� Hyamine (benzethonium chloride): very toxic[133]P34]
Medical Instrument Decontamination
After an invasive procedure or autopsy is performed, the instruments and area used should be disinfected with a sporicidal
1
agent, such as iodine or chlorine. Peracetic acid may also be used to decontaminate equipment and utensils.[351[136]
Proof of Decontamination
Proof of complete decontamination of soil and vegetation can be difficult because anthrax is a naturally occurring organism.
To decontaminate Gruinard Island (Scotland), a mixture of 283 tons of formaldehyde in 2,000 liters of seawater was
distributed over the area through a system of pipes. In 1988, fifty years after exposure, Gruinard Island was considered
anthrax-free.[I371
Russian Anthrax Compendium
(U/FOUO) This 1981 text entitled "Anthrax" by S.G. Kolesov presents exhaustive information from the height of the Soviet
era relating to the disease caused by Bacillus anthracis. Kolesov covers epidemiology, microbiology, detection and
treatment for this disease. At the time this book was published the Soviet biological warfare program was gearing up for
production of ton quantities of anthrax biological warfare agent. As a retrospective text for analysis this book provides
additional insights into Russian activities with anthrax. The book is divided into 3 parts [IPart I] , [I Part 2 ] ' [I Part 3 ]
Del 20:16, 28 August 2007 (UTC)
Anthrax and Botulinum Handbook, 1993
Summary
(U) This notebook consists of a series of tabs providing information on two biologics, anthrax and botulinum toxin; and on
seven technologies for identification of these biologies.
(U) The tabs provide information at two levels: an executive summary and at a detailed, more technical level of detail. In
addition, there is a reference section which contains tabs of useful reference material.
(U) Executive Level
� Tab A presents a summary, in matrix form, describing the capability of each of the seven selected identification
technologies to identify anthrax and botulinum. A synopsis explains the matrix entries. This tab summarizes the
material which follows in Tabs D and E.
� Tab B presents a generic summary of each of the seven identification technologies.
� Tab C presents a detailed write-up on botulinum and anthrax.
� Tab D presents work-ups of literature material in table form on the seven technologies capabilities applied to
identification of anthrax. There are two levels of detail: the higher level summarizes literature reviewed for all seven
biologies; the next level provides, for each technology cited in the upper level with more than two entries, specific
24 of 30 11/6/20189:29 AM
Approved for Release: 2021/11/22 C06785544
Anthrax - Intellipedia
Approved for Release: 2021/11/22 C06785544
(b)(3) 50 USC 3605
Doc ID: 6640699
amplifying information from each literature cite.
� Tab E presents work-ups of literature material in table form on the seven technologies
capabilities applied to identification of botulinum. Tab E contains material at the same level of detail as described above for
anthrax in Tab D.
� Tab F provides identification schemes for each biologic which provide both sufficient
and necessary justification for identification of the respective biologic. Detailed. Technical Level
� Tab G provides a detailed explanation for each of the technologies. Tab G is keyed to
items in Tab B.
� Tab H provides detailed abstracted information on each technology when used to identify each biologic; this material
was used to construct the tables in Tab D.
13 Tab I provides detailed abstracted information on each technology when used to identify each biologic; this material
was used to construct the tables in Tab E.
� Tab J contains additional schematic information amplifying Tab F. Tab J also discusses the differences in identification
schemes for environmental (dirty) samples vs for laboratory (clean) samples.
The Document is in 2 parts [I Part 1 , [I Part 2 ]
(U) Non-IC Resources
(U) The Department of Health and Human Services (HHS) is the lead Department for national threats to health, such as
anthrax. HHS's components include the Centers for Disease Control, which is the lead agency for public health, the National
Institutes of Health, which conducts extensive health-related research, and the Food and Drug Administration (FDA), whose
duties include approving potential countermeasures, vaccines, and protective gear that could be used in the event of another
anthrax attack.
(U) Please visit these organizations' Wiki pages to learn more about their interest in anthrax and for information on how to
contact their fully cleared intelligence interface staff.
Del 19:09, 11 September 2007 (UTC)
References
I. http://www.dia.ic.gov/intel/nuke/DI-CE/maceNCSPs/DragonWarrior
/ACTD/DTRAcY020Weapons%20of%20Mass.pdf WMD Terms Handbook
2. Serum Treatment of 19 Cases of Anthrax Including One of External, Internal and Bacteremic Type. Lucchesi, M. D.,
Pascal F. The American Journal of the Medical Sciences, 1952, pp. 796.
3. http://www.dia.ic.gov/intel/nuke/DI-CE/maceNCSPs/DragonWarrior
/ACTD/DTRA%20Weapons%200)/020Mass.pdf WMD Terms Handbook
4. A Brief Study of the Feasibility of Biological Testing Using Inert Simulants. Carlon, Hugh R. Aberdeen Proving
Ground, Maryland: Chemical Research Development and Engineering Center.
5. Some Thoughts on Biologicals as Biological Simulants. Bronk, B.V. Aberdeen Proving Ground, Maryland: Chemical
Research Development and Engineering Center, March 3, 1992.
6. Effectiveness of Medical Defense Interventions Against Predicted Battlefield Levels of Bacillus anthracis,. McNally,
Richard E.; Morrison, Malcom B.; Stark, M.; Bo'Berry, J.; Fisher, J. Joppa, Maryland: Science Applications
International Corporation, October 1, 1993, p. 4.
7. Soviet Biological Warfare Threat. Perroots, Leonard H. Washington DC: Defense Intelligence Agency, January 1,
1986, p.3.
8. FOA Informerar OM: A Briefing Book on Biological Warfare Agents.
25 of 30 11/6/2018 9:29 AM
Approved for Release: 2021/11/22 C06785544
Anthrax - Intellipedia
Approved for Release: 2021/11/22 C06785544
(b)(3) 50 USC 3605
Doc ID: 6640699
9. State-of-the-Art Report Biological Warfare Agent Detection Technologies. Bioarski, Anthony A.; Bowen, Gregory
W.; Durnford, Joyce M.; Kenny, Donald V.; Shaw, Matthew J. Aberdeen Proving Ground, Maryland: Chemical
Warfare/Chemical and Biological Defense Information Analysis Center, July, 1, 1995, pp. 3-4.
10. Diagnosis, Treatment, and Prophylaxis of Certain Potential Biological Warfare Diseases. Sutton, Leonard S.; Long,
William P.; Wedum, A. G. Frederick, Maryland: Army Biological Labs, January 1, 1954, p.6.
11. Joint CB Technical Data Source Book. Volume 8: Bacterial Diseases, Part 2-Anthrax. Fort Douglas, Utah: Desert Test
Center, February 1, 1973, p. 3-3.
12. Control of Communicable Diseases Manual. Benenson, Abram S. (Editor). Washington DC: American Public Health
Association, April 9, 1996
13. Chemical Warfare Agents. Somani, Satu M. San Diego, California: Academic Press, Inc., January I, 1992, p. 397.
14. Textbook of Military Medicine. Part I-Warfare, Weaponry, and the Casualty; Medical Aspects of Chemical and
Biological Warfare.Sidell, Takafuji, Franz (Editors). The Broden Institute.Washington DC: Walter Reed Army
Medical Center, June 1, 1997
IS. Effectiveness of Medical Defense Interventions Against Predicted Battlefield Levels of Bacillus anthracis,. McNally,
Richard E.; Morrison, Malcom B.; Stark, M.; Bo'Berry, J.; Fisher, J. Joppa, Maryland: Science Applications
International Corporation, October I, 1993, pp. A-3, 4.
16. Biological Warfare: Anatomy of a Terrorist Attack. Sizemore, Thomas D.; Zebovitz, Eugene. Columbus, Ohio:
Battelle Columbus Division, March I, 1991, p. 8.
17. Observations from Anthrax Outbreak in Sheridan County Herd. Gamby, John; Sahara, Roger; Grotelueschen, Dale.
University of Nebraska Extension. [Online], Available: http://www.ianr.unl.edu/PUBS/nebfacts/nf94-128.htm
18. Army Veterinary Research and Illustrated by Studies on Pathogenesis of Anthrax. Fitzpatrick, David F.; Klein,
Frederick; Lincoln, Ralph E.; Walker, Jerry S. Natick, Massachusetts: US Army Research Institute of Environmental
Medicine, January 1, 1963, pp. 4-5.
19. Introduction to Human Disease (Third Edition).
20. Crowley MD, Leonard V. Boston, Massachusetts: Jones and Barlett Publishers, 1992, p. 5.
21. Technology Against Terrorism: Structuring Security. Washington DC: Office of Technology Assessment, August 1,
1991 Cite error: Invalid tag; name "technologyagainst" defined multiple times with different content
Cite error: Invalid tag; name "technologyagainst" defined multiple times with different content
22. Effectiveness of Medical Defense Interventions Against Predicted Battlefield Levels of Bacillus anthracis. Morrison,
Malcom B.; Stark, M.; Bo'Berry, J.; Fisher, J.; McNally, Richard E. Joppa, Maryland: Science Applications
International Corporation, October 1, 1993, p. 5.
23. Technologies Underlying Weapons of Mass Destruction.Washington DC: Office of Technology Assessment,
December 1, 1993
24. Effectiveness of Medical Defense Interventions Against Predicted Battlefield Levels of Bacillus anthracis. Morrison,
Malcom B.; Stark, M.; Bo'Berry, J.; Fisher, J.; McNally, Richard E. Joppa, Maryland: Science Applications
International Corporation, October 1, 1993, p. 5.
25. Diagnosis, Treatment, and Prophylaxis of Certain Potential Biological Warfare Diseases. Sutton, Leonard S.; Long,
William P.; Wedum, A.G. Frederick, Maryland: Army Biological Labs, January 1, 1954, p.5.
26. Joint CB Technical Data Source Book. Volume 8: Bacterial Diseases, Part 2-Anthrax. Fort Douglas, Utah: Desert Test
Center, February 1, 1973, p. 3-3.
27. Diagnosis, Treatment, and Prophylaxis of Certain Potential Biological Warfare Diseases. Sutton, Leonard S.; Long,
William P.; Wedum, A.G. Frederick, Maryland: Army Biological Labs, January 1, 1954, p. 5.
28. Diagnosis, Treatment, and Prophylaxis of Certain Potential Biological Warfare Diseases. Sutton, Leonard S.; Long,
William P.; Wedum, A.G. Frederick, Maryland: Army Biological Labs, January 1, 1954, p. 5.
29. Diagnosis, Treatment, and Prophylaxis of Certain Potential Biological Warfare Diseases. Sutton, Leonard S.; Long,
William P.; Wedum, A.G. Frederick, Maryland: Army Biological Labs, January 1, 1954, p. 5.
30. Textbook of Military Medicine, Part I-Warfare, Weaponry, and the Casualty; Medical Aspects of Chemical and
Biological Warfare. Sidell Frederick R.; Takafuji, Ernest T.; Franz, David R.(Editors) Borden Institute, Washington
DC: Walter Reed Army Medical Center, June 1, 1997, p. 473.
31. Joint CB Technical Data Source Book. Volume 8: Bacterial Diseases, Part 2-Anthrax. Fort Douglas, Utah: Desert Test
Center, February 1, 1973, p. 3-3.
32. Observations from Anthrax Outbreak in Sheridan County Herd. Gamby, John; Sahara, Roger; Grotelueschen, Dale.
University of Nebraska. March 20, 1998, p. 1. [Online] Available: http://www.ianr.unl.edu/PUBS/nebfacts
/nf94-128.htm
33. Joint CB Technical Data Source Book. Volume 8: Bacterial Diseases, Part 2-Anthrax. Fort Douglas, Utah: Desert Test
Center, February 1, 1973, p. 3-3.
34. Observations from Anthrax Outbreak in Sheridan County Herd. Gamby, John; Sahara, Roger; Grotelueschen, Dale.
University of Nebraska. March 20, 1998, p. I. [Online] Available: http://www.ianr.unl.edu/PUBS/nebfacts
/nf94-128.htm
26 of 30 11/6/20189:29 AM
Approved for Release: 2021/11/22 C06785544
Anthrax - Intellipedia
Approved for Release: 2021/11/22 C06785544
(b)(3) 50 USC 3605
Doc ID: 6640699
35. Review of Medical Microbiology, Eleventh Edition. Jawetz, Ernest; Melnick, Joseph L.; Adelberg, Edward A. Los
Altos, California: Lange Medical Publications, 1974.
36. Textbook of Military Medicine. Part I � Warfare, Weaponry, and the Casualty; Medical Aspects of Chemical and
Biological Warfare. Sidell, Frederick R.; Takafuji, Ernest T.; Franz, David R. Borden Institute, Washington DC:
Walter Reed Army Medical Center, Junel, 1997, pp. 469-471.
37. Textbook of Military Medicine. Part I � Warfare, Weaponry, and the Casualty; Medical Aspects of Chemical and
Biological Warfare. Sidell, Frederick R.; Takafuji, Ernest T.; Franz, David R. Borden Institute, Washington DC:
Walter Reed Army Medical Center, Junel, 1997, pp. 469-471.
38. Technologies Underlying Weapons of Mass Destruction. Washington DC: Office of Technology Assessment,
December 1, 1993, pp. 93-97.
39. Possible Application of Biotechnology to the Development of Biological Agents by Potential Enemies. White,
William E. Aberdeen Proving Ground, Maryland: Chemical Research Development and Engineering Center, June 1,
1987, p.32.
40. Biological Warfare: Anatomy of a Terrorist Attack. Sizemore, Thomas D.; Zebovitz, Eugene. Columbus, Ohio:
Battelle Columbus Division, March 1, 1991, p. 5.
41. FOA Briefing Book. Ivarsson, Ulf; Nilsson, Helena; Santesson, Johan. UMEA, Sweden: National Defense Research
Establishment, January 1, 1992, p. 19.
42. Chemical Warfare Agents. Somani, Satu M., (Editors). San Diego, California: Academic Press, Inc., January 1, 1992,
p. 397.
43. Control of Communicable Diseases Manual. Benenson, Abram S. (Editor). Washington DC: American Public Health
Association, April 9, 1996, pp. 19.
44. Control of Communicable Diseases Manual. Benenson, Abram S. (Editor). Washington DC: American Public Health
Association, April 9, 1996, pp. 19-20.
45. Technologies Underlying Weapons of Mass Destruction. Washington DC: Office of Technology Assessment,
December 1, 1993, p.78.
46. Human Anthrax. Kadull, Paul J. Frederick, Maryland: Army Biological Labs, December 1, 1955, pp. 8-11.
47. The Military Impact on the Human Environment. Solna, Sweden: Stockholm International Peace Research Institute,
January I, 1978, p.49.
48. Technologies Underlying Weapons of Mass Destruction. Washington DC: Office of Technology Assessment,
December 1, 1993, p.78.
49. Effectiveness of Medical Defense Interventions Against Predicted Battlefield Levels of Bacillus anthracis. Morrison,
Malcom B.; Stark, M.; Bo'Berry, J.; Fisher, J.; McNally, Richard E. Joppa, Maryland: Science Applications
International Corporation, October 1, 1993, pp. A-3, 4.
50. Control of Communicable Diseases Manual (Sixteenth Edition). Benenson, Abram S., Editor. Washington DC:
American Public Health Association, April 9, 1996, p. 20.
51. Textbook of Military Medicine, Part I-Warfare, Weaponry, and the Casualty; Medical Aspects of Chemical and
Biological Warfare. Sidell, Frederick R.; Takafuji, Ernest T.; Franz, David R., Editors. Borden Institute, Washington
DC: Walter Reed Army Medical Center, June 1, 1997, p.474.
52. Defense LINK News Release No. 094-98. [Online], Available: http://www.defenselink.mil/news. March 3, 1998.
53. Control of Communicable Diseases Manual (Sixteenth Edition). Benenson, Abram S., Editor. Washington DC:
American Public Health Association, April 9, 1996, p. 20.
54. Textbook of Military Medicine, Part I-Warfare, Weaponry, and the Casualty; Medical Aspects of Chemical and
Biological Warfare. Sidell, Frederick R.; Takafuji, Ernest T.; Franz, David R., Editors. Borden Institute, Washington
DC: Walter Reed Army Medical Center, June 1, 1997, pp. 473-475.
55. Effectiveness of Medical Defense Interventions Against Predicted Battlefield Levels of Bacillus anthracis.
56. McNally, Richard E.; Morrison, Malcom B.; Stark, M.; Bo'Berry, J.; Fisher, J. Joppa, Maryland: Science Applications
International Corporation, October 1, 1993, p. 6.
57. Plant Design for Large-scale Production of an Anthrax Immunizing Antigen. Edgewood Arsenal, Maryland: Director
of Engineering and Industrial Services, January 1, 1964, p. I.
58. Possible Application of Biotechnology to the Development of Biological Agents by Potential Enemies. White,
William E. Aberdeen Proving Ground, Maryland: Chemical Research Development and Engineering Center, June 1,
1987, p.33.
59. Textbook of Military Medicine, Part I-Warfare, Weaponry, and the Casualty; Medical Aspects of Chemical and
Biological Warfare. Sidell, Frederick R.; Takafuji, Ernest T.; Franz, David R., Editors. Borden Institute, Washington
DC: Walter Reed Army Medical Center, June 1, 1997, pp. 473-475.
60. Textbook of Military Medicine, Part I-Warfare, Weaponry, and the Casualty; Medical Aspects of Chemical and
Biological Warfare. Sidell, Frederick R.; Takafuji, Ernest T.; Franz, David R., Editors. Borden Institute, Washington
DC: Walter Reed Army Medical Center, June 1, 1997, p.474.
27 of 30 11/6/2018 9:29 AM
Approved for Release: 2021/11/22 C06785544
Anthrax - Intellipedia
Approved for Release: 2021/11/22 C06785544
(b)(3) 50 USC 3605
Doc ID: 6640699
61. Effectiveness of Medical Defense Interventions Against Predicted Battlefield Levels of Bacillus anthracis. McNally,
Richard E.; Morrison, Malcom B.; Stark, M.; Bo'Berry, J.; Fisher, J. Joppa, Maryland: Science Applications
International Corporation, October 1, 1993. p. 6.
62. Expression of Cereolysine AB Genes in Bacillus anthracis Vaccine Strain Ensures Protection Against Experimental
Hemoytic Anthrax Infection. Pomerantsev, A.P; Staritsin, N.A.; Mockov, Yu.V.; Marinin, L.1. Telaviv, Israel: Sackler
School of Medicine, January 1, 1997, Volume 15 No. 17/18, p. 1850.
63. Effectiveness of Medical Defense Interventions Against Predicted Battlefield Levels of Bacillus anthracis. McNally,
Richard E.; Morrison, Malcom B.; Stark, M.; Bo'Berry, J.; Fisher, J. Joppa, Maryland: Science Applications
International Corporation, October 1, 1993, p. 6.
64. Textbook of Military Medicine, Part I-Warfare, Weaponry, and the Casualty; Medical Aspects of Chemical and
Biological Warfare. Sidell, Frederick R.; Takafuji, Ernest T.; Franz, David R., Editors. Borden Institute, Washington
DC: Walter Reed Army Medical Center, June 1, 1997, p. 475.
65. Textbook of Military Medicine, Part I-Warfare, Weaponry, and the Casualty; Medical Aspects of Chemical and
Biological Warfare. Sidell, Frederick R.; Takafuji, Ernest T.; Franz, David R., Editors. Borden Institute, Washington
DC: Walter Reed Army Medical Center, June 1, 1997, p.473.
66. Chemical Warfare Agents. Somani, Satu M., Editor. San Diego, California: Academic Press, Inc., 1992, pp. 397, 399.
67. 9. Anthrax as a Biological Warfare Agent. [Online], Available: http://www.defenselink.mil/other_info/agent. March
11, 1998. Joint CB Technical Data Source Book. Volume 8: Bacterial Diseases, Part 2-Anthrax. Fort Douglas, Utah:
Deseret Test Center, February 1, 1973, pp. 4-7.
68. Biohazard. Alibek, Ken. Random House. New York, 1999. pp. 160, 167.
69. Joint CB Technical Data Source Book. Volume 8: Bacterial Diseases, Part 2-Anthrax. Fort Douglas, Utah: Deseret
Test Center, February 1, 1973, pp. 4-7.
70. Textbook of Military Medicine, Part 1-Warfare, Weaponry, and the Casualty; Medical Aspects of Chemical and
Biological Warfare. Sidell, Frederick R.; Takafuji, Ernest T.; Franz, David R., Editors. Borden Institute, Washington
DC: Walter Reed Army Medical Center, June 1, 1997, p.473.
71. Diagnosis, Treatment, and Prophylaxis of Certain Potential Biological Warfare Diseases. Sutton, Leonard S.; Long,
William P.; Wedum, A.G. Frederick, Maryland: Army Biological Labs, January 1, 1954, p. 4.
72. Effectiveness of Medical Defense Interventions Against Predicted Battlefield Levels of Bacillus anthracis. McNally,
Richard E.; Morrison, Malcom B.; Stark, M.; Bo'Berry, J.; Fisher, J. Joppa, Maryland: Science Applications
International Corporation, October 1, 1993, p. 7.
73. Chemical and Biological Contamination Avoidance. FM 3-3. Washington DC: Headquarters, Department of the
Army, Navy and the US Marine Corp., November 16, 1992, p. A-4.
74. NBC Field Handbook. Washington DC: Department of the Army, September 29, 1994, pp. 55-56.
75. Effectiveness of Medical Defense Interventions Against Predicted Battlefield Levels of Bacillus anthracis. McNally,
Richard E.; Morrison, Malcom B.; Stark, M.; Bo'Berry, J.; Fisher, J. Joppa, Maryland: Science Applications
International Corporation, October 1, 1993, p. 7.
76. Effectiveness of Medical Defense Interventions Against Predicted Battlefield Levels of Bacillus anthracis. McNally,
Richard E.; Morrison, Malcom B.; Stark, M.; Bo'Berry, J.; Fisher, J. Joppa, Maryland: Science Applications
International Corporation, October 1, 1993, p. 7.
77. Chemical Warfare Agents. Somani, Satu M., Editor. San Diego, California: Academic Press, Inc., 1992, p. 399.
78. Joint CB Technical Data Source Book. Volume 8: Bacterial Diseases, Part 2-Anthrax. Fort Douglas, Utah: Deseret
Test Center, February 1, 1973, pp. 4-7.
79. Textbook of Military Medicine, Part 1-Warfare, Weaponry, and the Casualty; Medical Aspects of Chemical and
Biological Warfare. Sidell, Frederick R.; Takafuji, Ernest T.; Franz, David R., Editors. Borden Institute, Washington
DC: Walter Reed Army Medical Center, June 1, 1997, p.473.
80. Control of Communicable Diseases Manual (Sixteenth Edition). Benenson, Abram S., Editor. Washington DC:
American Public Health Association, April 9, 1996, p.21.
81. Human Anthrax. Kadull, Paul J. Frederick, Maryland: Army Biological Labs, December 1, 1955, p. 17.
82. Observations from Anthrax Outbreak in Sheridan County Herd. Gamby, John; Sahara, Roger; Grotelueschen, Dale.
University of Nebraska Extension. [Online], Available: http://www.ianr.unl.edu/PUBS/nebfacts/nf94-128.htm March
20, 1998.
83. Effectiveness of Medical Defense Interventions Against Predicted Battlefield Levels of Bacillus anthracis. McNally,
Richard E.; Morrison, Malcom B.; Stark, M.; Bo'Berry, J.; Fisher, J. Joppa, Maryland: Science Applications
International Corporation, October 1, 1993, pp. 3, 6-7.
84. Diagnosis, Treatment, and Prophylaxis of Certain Potential Biological Warfare Diseases. Sutton, Leonard S.; Long,
William P.; Wedum, A.G. Frederick, Maryland: Army Biological Labs, January 1, 1954, pp. 4-5.
85. Harvard Researchers Put Anthrax Toxin to Good Use. Collier, John. Harvard University. February 19, 1998. (Online),
Available: http://www.eurekalert.org/releases/anthraxtox.html
86. Harvard Researchers Put Anthrax Toxin to Good Use. Collier, John. Harvard University. February 19, 1998. (Online),
28 of 30 11/6/2018 9:29 AM
Approved for Release: 2021/11/22 C06785544
Anthrax - Intellipedia
Approved for Release: 2021/11/22 C06785544
(b)(3) 50 USC 3605
Doc ID: 6640699
Available: http://www.eurekalert.org/releases/anthraxtox.html
87. Harvard Researchers Put Anthrax Toxin to Good Use. Collier, John. Harvard University. February 19, 1998. [Online],
Available: http://www.eurekalert.org/releases/anthraxtox.html
88. Harvard Researchers Put Anthrax Toxin to Good Use. Collier, John. Harvard University. February 19, 1998. [Online],
Available: http://www.eurekalert.org/releases/anthraxtox.html
89. Biological Warfare: Anatomy of a Terrorist Attack. Sizemore, Thomas D.; Zebovitz, Eugene. Columbus, Ohio:
Battelle Columbus Division, March 1, 1991, Table 6.
90. Selection of a Seed Stock for Bacillus Anthracis with Emphasis on Factors Affecting Colonial Variation. Klien,
Frederick; Matcalfe, Nicholas S., Jr. Frederick, Maryland: Army Biological Labs, November 30, 1956, p. 77.
91. Biological Warfare: Anatomy of a Terrorist Attack. Sizemore, Thomas D.; Zebovitz, Eugene. Columbus, Ohio:
Battelle Columbus Division, March 1, 1991, p. 71
92. Evaluation of Media for Growth of Bacillus anthracis and Pasteurella tularensis. Wolfe, Victor L.; Klien, Frederick.
Fort Detrick, Maryland: Biological Warfare Laboratories, Aprill, 1969.
93. Biological Warfare: Anatomy of a Terrorist Attack. Sizemore, Thomas D.; Zebovitz, Eugene. Columbus, Ohio:
Battelle Columbus Division, March 1,1991, p. 74.
94. Biological Warfare: Anatomy of a Terrorist Attack. Sizemore, Thomas D.; Zebovitz, Eugene. Columbus, Ohio:
Battelle Columbus Division, March 1, 1991, pp. 71, 74, 94, Table 6.
95. Joint CB Technical Data Source Book. Volume 8: Bacterial Diseases, Part 2-Anthrax, Fort Douglas, Utah: Deseret
Test Center, February I, 1973, pp. 4-7.
96. Technical Aspects of Biological Defense. Washington DC: Departments of the Army and the Air Force, January 1,
1971, p.57.
97. Biological Defense Operations, Corps/Company Tactics, Techniques, and Procedure. Washington DC: Headquarters
Department of the Army, August 1, 1997, pp. 1-4, 1-5.
98. Medical Management of Biological Casualties Handbook. Second Edition. Fort Detrick, Maryland: Army Medical
Research Institute of Infectious Diseases, August 1, 1996.
99. Assessment of Biological Agent Detection Equipment for HAZMAT Responders to US Army Chemical and
Biological Defense Command. Barrett, John A.; Bowen, Gregory W.; Golly, Scott M.; Hawley, Chris; Jackson,
William M.; Laughlin, Leo; Lynch, Megan E Bel Air, Maryland: Battelle, March 1, 1998.
100. State-of-the-Art Report Biological Warfare Agent Detection Technologies. Bioarski, Anthony A.; Bowen, Gregory
W.; Durnford, Joyce M.; Kenny, Donald V.; Shaw, Matthew J. Aberdeen Proving Ground, Maryland: Chemical
Warfare/Chemical and Biological Defense Information Analysis Center, July 1, 1995, pp. 8-9. Technology against
Terrorism: Structuring Security. Washington DC: Office of Technology Assessment, August 1, 1991, p.37.
101. State-of-the-Art Report Biological Warfare Agent Detection Technologies. Bioarski, Anthony A.; Bowen, Gregory
W.; Durnford, Joyce M.; Kenny, Donald V.; Shaw, Matthew J. Aberdeen Proving Ground, Maryland: Chemical
Warfare/Chemical and Biological Defense Information Analysis Center, July 1, 1995, pp. 8-9, 42, 47.
102. Medical Management of Biological Casualties Handbook. Second Edition. Fort Detrick, Maryland: Army Medical
Research Institute of Infectious Diseases, August 1, 1996.
103. State-of-the-Art Report Biological Warfare Agent Detection Technologies. Bioarski, Anthony A.; Bowen, Gregory
W.; Durnford, Joyce M.; Kenny, Donald V.; Shaw, Matthew J. Aberdeen Proving Ground, Maryland: Chemical
Warfare/Chemical and Biological Defense Information Analysis Center, July 1, 1995, pp. 8-9, 42, 47. Medical
Management of Biological Casualties Handbook. Second Edition. Fort Detrick, Maryland: Army Medical Research
Institute of Infectious Diseases, August 1, 1996.
104. Development of Enzyme-Linked Immunosorbent Assay (ELISA) to Anthrax for the Persian Gulf. Nagata, Les P.;
Schmaltz, Fay L.; Balogh, Carol; Bhatti, A Rashid; Cherwonogrodzky, John W. Suffield Ralston, Alberta: Defense
Research Establishment, July 1, 1995, p. 1.
105. State-of-the-Art Report Biological Warfare Agent Detection Technologies. Bioarski, Anthony A.; Bowen, Gregory
W.; Durnford, Joyce M.; Kenny, Donald V.; Shaw, Matthew J. Aberdeen Proving Ground, Maryland: Chemical
Warfare/Chemical and Biological Defense Information Analysis Center, July 1, 1995, pp. 8-9, 17, 53.
106. Medical Management of Biological Casualties Handbook. Second Edition. Fort Detrick, Maryland: Army Medical
Research Institute of Infectious Diseases, August 1, 1996.
107. Deadly Secrets. Hinckle, W.; Turner, W.
108. Proliferation of Weapons of Mass Destruction. Washington DC: Office of Technology Assessment, August 1, 1993, p.
49.
109. Biological Warfare: An Overview. Patrick, William C. Frederick, Maryland: William C. Patrick, May 1, 1994, p.3.
110. Chemical and Biological Warfare Investigation Guide. pp. 82-85. Background and Persistency Studies for Agent N.
BW 10-54 Operation "Dust Mote." July 13, 1955.
Ill. Biological Warfare: An Overview. Patrick, William C. Frederick, Maryland: William C. Patrick, May 1, 1994, p. 3.
112. Effectiveness of Medical Defense Interventions Against Predicted Battlefield Levels of Bacillus Anthracis. Morrison,
Malcom B.; Stark, M.; Bo'Berry, J.; McNally, Richard E. Joppa, Maryland: Science Applications International
29 of 30 11/6/2018 9:29 AM
Approved for Release: 2021/11/22 C06785544
Anthrax - Intellipedia
Doc 1D: 6640699
Approved for Release: 2021/11/22 C06785544
(b)(3) 50 USC 3605
Corporation, October 1, 1993, p. 15.
113. Biological Warfare: An Overview. Patrick, William C. Frederick, Maryland: William C. Patrick, May 1, 1994, p. 3.
114. Medical Management of Biological Casualty Handbook. Second Edition. Fort Detrick, Maryland: Army Medical
research Institute of Infectious Diseases, August 1, 1996, p.2.
115. Biological Warfare: An Overview. Patrick, William C. Frederick, Maryland: William C. Patrick, May 1, 1994, p. 3.
116. Proliferation of Weapons of Mass Destruction. Washington DC: Office of Technology Assessment, August 1, 1993, p.
49.
117. Medical Management of Biological Casualty Handbook. Second Edition. Fort Detrick, Maryland: Army Medical
research Institute of Infectious Diseases, August 1, 1996, p.2.
118. Biological Warfare: An Overview. Patrick, William C. Frederick, Maryland: William C. Patrick, May 1,1994, p.3.
119. Human Anthrax. Kadull, Paul J. Frederick, Maryland: Army Biological Labs, December 1, 1955.
Retrieved from
(b)(3) 50 USC 3605
Categories: Pages with reference errors I History of Intelligence I Anthrax I Biological Agents I Epidemiology I Health
Biological warfare I Dictionary I WMD
Classified By:
Derived From:
Declassify On June 17, 2041
CON Fl
(b)(3) CIAAct
(b)(3) NatSecAct
(b)(3) 50 USC 3605
^ This page has been accessed 7,556 times. (b)(6)
� 12 watching users
1:3 This page was last modified 08:05, 17 June 2016 by Most recent editors
and and others. (b)(3) CIAAct
(b)(3) CIAAct (b)(3) Nat5ecAct9;
(b)(3) NatSecAct
Use of this U.S. Government system, aulhorbikwa vthorized, constitutes consent to monitoring of this system. Unauthorized use may subject( b)(6)1inal prosecution.
Evidence of unauthok Li) kL/Lollected during monitoring may be used for adminbtrative, criminal, or other adverse actions.
TOP SECRET//HCS-P/SI-G/TK//NOFORN/ORCON
30 of 30
Approved for Release: 2021/11/22 C06785544
11/6/20189:29 AM