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Proiect No. l-X-6-65704-D-Ll4 Proiect No. 5-C0-473-000-026 DTC Proiect No. DTC-TR-73-517

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L]OINT CB TECHNICAL DATA SOURCE BOOK tU]

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VOLUME VI I
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Bacterial

Diseases (U)
(U)

:\)

Part Two: Anthrax

FEBRUARY 1973
(J

a /j:,-araz ler:'; zli'L

HEADQUARTERS

'

DESERET TEST CENTER

'

FORT DOUGLAS,

UTAH

O 84I I3

DTC 73-27

Copy

'' of 5 CoPies

I]NCLASSIFIED
(u)
r'oREwoRD (u)

This document was prepared in compliance with Department of the Army letter, I'Joint contacE Point for chemical-Biological- (cB) Fiel_d Test Data (u) r" 10 March L967, r.rhich direcred Deseret Test center (DTc) to pubLish and maintain a joint chemical-biological weapons system
source book. The Source Book is organized into volumes by agent category. In each vol-ume, Parameter val-ues and confidence levels derived. from field, laboratory, and chamber test data are presented. This volume also Presents modeLs and submodels that identify and define the parameters for which numerical values are required in estimating the capabilities of weapons systerns. The weapons systerns for which data are presented incl-ude those of each of the Armed Services that had been standardized or t)rye-classified and Ehose that were in an advanced stage of development prior to the presidential poliey statement of 25 Novernber 1969 regarding

t
a

'f
F l:

,
F

biological weapons.

The material presented in each volume is organized in such a manner that it can be used by (1) the research and development corununity as input to system design and analysis studies and (2) the operational conrnunity as input to the Preparation of system-performance tabLes for inclusion in field nnnuals, firing tabl-es, and other presentations of munitions expenditure and effectiveness information.

The assistance of GEOMET, Incorporated, who prepared major portions of this document under DA ContracE No. DAAD 09-69-C-0078, is ackknowledged.

and suggesEions regarding the adequacy or accuracy of the material presented herein and any assistance needed in its use should addressed to:
Comments
Commander

be

Deseret Test Center ATTN: STEPD-PS-A(S) Fort Douglas, UEah

84113

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F .'

I]NCI,^ASSIFIED

l--

IINCIJ.SSIFIED
(u)
Fiqure
3-1
1

TLLUSTRATTONS (U)
Pa

ge

Response of Cynomolgus Anthracis.....

Monkey

to Aerosols of

B. 3- 10

3-Z 4-L
5-1

r-1

Total and Regional Deposition of Inhaled Particles in Relation co Aerodynamic Particle Size.
Freeze-Thaw Cycling Tests in the TRI Logistic Study

3-L2
4- 10

,L
-1
L

Concentration of Viable Bacillus Anthracis Spores in Aerosols Maintained in the.Dynamic Aerosol Toroids at Four Relative HumidiEy Percentages and 27 oC. for a Period of 138 Hours
TRI Downwind Dosage Predictions for Aerial Releases During Windspeeds of 3 Meters per Second.

5-6
6-2

6-1

-1
6-2
6-3

TRl Downwind Dosage Predictions for Aerial Releases During Windspeeds of 6 lleters per Second. . During Windspeeds of 3 Meters per Second.
TR2 Dc'ranwind Dosage

6-3
6-4

Predictions for Aerial Releases

-l
6-4
.6

TR2 Downwind Dosage PredicLions for Aerial Releases During Windspeeds of 6 Meters per Second.

6-5

lx

TTNCI.^A''SSIFIED

IINCLASStiEI
(u)
Number
TABLES
?a ge

4-L
/,

Initia I Concentrations fc: i= - -.::=: I -=Forc Detrick Aerosol Tests.

TR1

. -

4_2

_')

Harvest ConcenEraEions and tsu-; : -:::::- -: -Composite Lots of DBO-Produce: l-Decay Constants and Half-Life. Various Temperatures.

4-3 4-5 4-6

4-3

::: I

: -:=----

4-4
4-5

Estimated Storage Decay Pare;-^-:::--= : :: =i =r-- \rariations in Guinea Pig Resp---=::-=- --i: -:: Stored f or Different Time Per::,:.= :: -==-.r:lures

4-9

4-6
4-7

Viable Spore Millilirer x

Count.

10s)

!.-LL

E6lR4 Bomblet Dissemination E::::- =.::- - : ..;.=- - -:as a Function of Time and Te=:=-:=: =: --- ,;:-Storage
' j--:-=--r: --=-r TotaI Aerosol Decay Rate f or i:::=-- -:l-=== from che E61R4 Bomblet in the I::: Function of Time and Temperai -:: :: -i'-; . - :- -=-

4-L3

4.- 8

4I

13

4-9
4-10
L-1

Characleristics of }lunitions l::=: -= :: - -- :-r1---:r-=i B. AnEhracis.

Dissemination Efficiencies cf Tes t Ct"rambers
Estima:ed TR2 Disseminaiion E::: //10 Nozzle with Tornado Feece:

L7

;-

18

:- 18
:--20

/,

-1a

Efficiencl' of Dissemina tion c: and A/845y-4 Spray Tanks.

Estimated Dissemination Systemrs with Agent TR
Ef

4-

l3

fic-

:-zL
=t

5-1_

Estinated Total Decay Rates (ToLaI Decey Ra tes f or the I'{iCharnbe:: Tes ts
-

5-2

I'NCI.^A,SSE

T'NCI,^ASSIFIED
(u)
e

TABLES

(U) (Continued)
Pa ge

Number

5-3
.3

Total Decay Rates (100k) for Trials at Fort Detrick . .

TR2 Based on Chamber

5-4
5-7

5-4

Biological Decay Rates (Percent/Minute) for TRl under Ultraviolet Stress

BG and TRl Total Decay E61R.4 Bomblets

l
i

1
I

5-5 6-7

Rates from Chamber Tests Using

5-8

Estimated Downwind Distances for Effective Dosage (EED = 5.3 x 106 org rnin/m3) of TRl and TR2 Released from an Aerial Line Source
Recommended Chemotherapy Regimens

6-7

7-L
7-2

for Anthrax.
B.

7-3
7-9 7-LL
7-L2

Effects of Various Disinfectants on the Spores of AnEhracis Suspended in Solution. .

Formaldehyde Gas Srerilization and Equipment.

l-3
7-4
7-5

of FaciliLies, Materials,

Characteristi cs of Selected Decontaminants

Amounts of Be rapropiol-actone (BPL) Released in Ship Experiments.

-L2

X1

UNCIJ.SSIFIED

CHAPTER 3

(s)

AGENT Cr{AMCTERTSTTCS (u)

(U) Biological Nature of the Organism

a. Morphological Characteristics. Bacillus anthracis is an aerobic, sporulating, gram-posiEive, nonmotile, rod-shaped bacterium. Cel1s may vary in size from 1 to 1.5 by 3 to 10 microns, depending on the conditions of growth. Cells sporul-ate readily when gror.m in laboratory culture, but they do not sporulate when in living tissue. Presumably, sporulation occurs in cells grown in living animal tissue when the cell-s are discharged from the animal body inEo a suitable medium such as soil or organic residues. Spores are ellipsoidal to cylindrical, 0.8 to 1.0 by 1.3 to 1.5 microns, and they occupy a median position in the cell-. The cells are usually in chains and have square ends. Cells thaE grovr in the animal body or laboratory culture in an atmosphere containing 10 to 15 percent ( CO" form capsules. L'2,3)
b. Cultural Characteristics. B. anthracis requires free oxygen for growEh and spore formation; it is therefore considered an aerobe. It grows well on most cultural media at temperatures ranging from 12 to 45 oC. B. anthracis is generally nonhemolyEic. Typically, colonies of virulent strains of the organism, when grown on nutrient agar, are greyishwhite in color and have hair-like tufts around the peripherial surface of Ehe colony (these are sometimes referred to as medusa-head-type co1-onies). The surface of the colony is granul-ar in appearance as compared to the usually small, round, smooth-cype colonies of the avirulent strains. However, when viruLenE organisms are grown on special media such as serum agar in Ehe presence of a high CO. atmosphere, smooth colonies of virulent encapsulated cells are formed. Encapsulation has been associated with virulence of the organism; however, this association may be fortuitous. Encapsulation has not been directly associated with the ability of the organism to prcduce toxin. Encapsulation is relaEed Eo the resistance of the B. ant.hracis ce11 to in" action of opsonin, an anEibody associaEed with phagocytos:s. Avirulent mucoid muEanEs have been described. These variant.s are encapsulated when grown on nutrienE agar in air. Asporogenous variants have also been ceveloped which may be eiEher virulent or avirulent.(1-4) c. Persistency. The anEhrax bacillus can persist in the sgore form indefinit"lyl" rriT and organic matter, particularly if the environmenL is dry. The vegetaEive organism may also grow and reproduce in soil and organic materia-s. Hcrqrever, vegetative ce1ls of B. anthracis do not survive well in :omperition with the saprophytic o?StrGsE tt.r. so!i, and in some sclls the organism does not become esEablished as an indigenous member of ihe soil microflora. However, the organism occurs enieni.cally

in many areas of the world as a natural soil inhabitant.(].t1ts) The organism survives for long periods on various animal products, such as hides, wool, hair, bone meal, and meat product.s, and it may be transported by commerce in these materials
Ir

3-3. (U) Characteristics of the Disease a. Ncmenclature. Anthrax is preCominantly a disease of animals (catt1e, sheep, horses, and swine). In animals, Lhe disease is also knor,in by such names as splenic fever, charbon, and milzbrand. The disease may exist in man as a malignant pustule (cutaneous anthrax), as inhalation anthr:x, or as intestinal anthrax. Inhalation anthrax (frequently referred tc as pulmonary anthrax) most frequently occurs among workers in industrial plants where dust from processed materials contaminated wich B. anthracls enters the respiratory system of the worker. Because of its freque:it occurrence in workers in specific jobs, it is kncnrn by such names as wcolsorter I s disease and ragsorter t s disease. lntestinal anthrax results fr--m ingestion of the sPores. It has been reported in humans, though inf:equently. It is relativel-y ccrTunon in animals, whereas inhalatj-on anthr:x infrequenLly occurs in aninals. Cutaneous anthrax or malignant pustule is the most frequent forirr of t:re disease in both anim:1s
and humans,

J-:

UNCTASSIFIED
b. Transmission. AnEhrax is generally transmitted to man from animals or conEaminaEed material associaEed with animals. No man-to-man transmission has been reported. Many mammals are suscepEible to anthrax as a result of natural infection or by laboratory-induced infection. Birds are relatively refracEory; however, ant,hrax has been reported in some carnivorous birds and in the osErich. Scavenging birds may carry and disperse t,he organism. Cold-blooded animals are generally refractory Eo Ehe disease; however, css have been reported in frogs and some fish. The cuEaneous form of the disease occurs when the anthrax organism enters the hosE through an abrasion in the skin and forms a lesion at the surface of the skin at the point of enEry. Early historical reports of ragsorterts disease and woolsorterrs disease indicated fairly frequent occurrence. However, improvements in sanitaEion and methods of handling animals and animal producEs have conEr.ibuted to a reduction in inhalation anthrax. As an internal infection, this disease may occur after ingestion of contaminated material or by inhalation of cont.aminated dust or othenrise aerosolized B. anthracis organisms. The disease is not considered infect,ious, because iE is not easily nor readily Eransmitted, as evidenced by the widespread occurrence of the organisms in the soil in rnany areas of the world with low incidence of the disease. Horve'rer, in enzootic areas, Large-scale epidemics among animal populations have occurred. The disease is most commonly transmitEed in animals by ingestion of contaminated food. Some experimental work has indicated Ehat the organism may be transmitted from hosE to host by flies, mosquitos, and oEher vecEors. ( I '4-6)
Svmptoms and Diagnosis

(1) Cutaneous anthrax is characterized by a local lesion which begins from a small red nncule and enlarges to form a central vesicle of clear fluid surrounded by satellite vesicles. The lesion becomes necrotic, and a black eschar is characteristic of older lesions. The Iesion is not painful, but the regional lymph nodes are Eender. The organism may be isolated from the lesion, parEicularly during early developrnenE. Malaise, fever, headache, and general prostraEion develop, and the intensiEy of the symptoms vary with the hosE, the stage of Ehe disease, and apparently tviEh Ehe number of organisms involved in the initial infection, i.e., the dose. In the early stage, pulmonary and intestinaL anchrax symptorns are mild and nonspecific. This early stage is followed by a very rapid onsec of che advanced disease, associated wit.h a nassive invasion by the organism throughout Che body.(e) (2) The two stages of human inhalation ant.hrax described below are characterized as insidious onset and acuEe toxemia. (a) Insidious onset is associated with mild fever, malaise, fatigue, Ryslgia, a nonproducti-ve cough, and frequenElv a sensaEion of precordial oppression. This inicial stage typically lasts for several <ieys . Tie pa tient's clinj-ca1 co:rdir:on may irnprcve slightiy rowarC the end of !ris stage.(a)
)- a

UNCLASSIFIED
L----

I'NCTASSIFTED
(b) Acute toxemia develops suddenly, with acute dyspnea (shorcness of breath) and subsequent cyanosis (1ack of blood at the body surface). The patient aPPears moribund, with accelerated pulse and respiration. The body Eemperature, although usually elevated to 102 oF. or more, ffiy suddenly become subnormal because of shock. Profuse perspiration commonly occurs. Subcutaneous edema of chest and neck may exist. Stridor is common, and chest examination discloses crepitant rales associated with fluid in the lungs.(6'a'e) The average duration of this acute stage is less than 24 hours, and it terminaEes in death. Consciousness is usually nEintained until death, except v/ith the infrequent occurrence of meningitis, when disorientation and coma occur. (3) Since the clinical manifestations of inhalation and intestinal anthrax are nonspecific and inconsistent, diagnosis of the disease remains a major problem. Diagnosis of inhalation anthrax can be aided by the following (") A history of occupational exposure by natural infection.(1o) (Plotkin, et al.r(8) suggest that rreatment should be started on the basis of suspicion or association rather than on positive diagnosis.) of the mediastinum. (b) Roentgenographic examination revealing acute widening
(8 1o '
)

(.) Positive blood cultures indicating disseminated anthrax infection; occasionally the bacilli may be identified in the centrifuged sediment of blood treated with 3-percent aceEic acid solution and stained with Wrightts stain.(10) Hot",r"t, if man reacts in a manner similar to monkeys and chimpanzees, septicemia may be deEected by blood smear examination about 10 hours letore death.(ro)
I I

(4) The following laboratory methods for ver]- fication of anthrax infection in i11 or dead animals may be employed .(rt) (") Direct microscopic examination of suspected material, when stained. satisfactorily, will reveal gram-positive bacilli that are 1 to 1.5 microns in diameLer and 5 io B microns 1ong. In blood smears, most of the bacilli will be single cel1s, but short chains may exist if the animal has been dead for a few hours. (b) Inoculation of tryPtose sov agar plates with infected will show medusa-headed colonies in 12 to 24 hours. blood (") Animal inoculation in guinea pigs or mice is extremely va1uable. liinety-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.

i
i i

I I

I I
I I

I
I

I
I

3-4

TINCIASSIFIED

ITNCLASSIFIED
(d) Fluorescent antibody techniques, which work equally well blood smears and tissue sections , can be used for rapid identification on of B. anEhracis.
menE

d. Onset and Duration. Following cutaneous infection, developof Ehe lesion usually occurs within 2 ot 3 days. If treacment is iniE iated, Ehe infection may spread systemically co bacterimic pronot porEions and death may occur within 4 to 7 days. For inhalation anthrax !n humans, Ehe incubation period is said to be as short as 24 hours(a) and as long as from 4 to 5 days.(s) For man, death resulting from inha 1a tion anthrax occurs from 13 hours to 12 days folLcnuing onset of symptoms.lrz) For a recent single human case of inhalation anEhrax, the date of exposure \{as fairly accurately idenEified. OnseE of the disease for this case e/as approximately 6 days to insidious manifestation of illness and 10 days to acute illness and death.(13) Monkeys, exposed under similar circumstances, had onset times of 10 to L7 days.(r+)
I'lechanism of Infection

(1) The mechanism(s) oi infection by B. anthracis is not completely understood,,!yt iq aPPears to be a quantitaLive chemical propercy of the agent.(o'J Virulence of B. anthracis may Possibly be associated with particular cellular antigens, with the ability or inability of the anthrax organism to be phagocytized by the host macrophages, with the abilicy or inability of phagocytic ce1ls to detoxify Ehe agent,, or wifh Che ability of the agenc to reproduce within these ce11s. No data have been found co indicaEe that the material from which the spore was derived, i.e., !/eE material or dry maEerial, has any effecE on the infective process. (2) Experiments seem to indicate that the vegetative ce11 injected inEo Ehe body kills the guinea pig in a shorter time than the spore. While in Ehe germination stage, the ce11 probably develops an active, integrated physiology and remains noncapsulated- This is probably Che period of its greatest susceptibility to destruction- The encapsul-ated cel1 is protected to some degree by the polyglutamic acid capsule. Anthracidal substances in the blood appear to be the principal in the blood, and phagocytosis means c destruction of anthrax bacilli (" ) The first noticeable effect of anthracidal is of iess importancu. nnterial on the bacillus is a loss of capsule, accompanied by a loss of sEaini::g Droperties, followed by fragmentation. AcEually, t'anthracidal subsE.ancest' is a general term embracing many facEors. Dog organs contain a decacsulating agent, and there seems to be both a heat-stable and a heat-la'oile anthracidal factor in guinea pig Leucocytes. (3) PaEhogenes is . of i-nha lation anthrax has three distinct srages: the prr:;rary stege, during which s?ores nove from the lung alveoli into the an::nal body and establish a primary infection in the lymph syst'm; a per:--C of secondary grcwth in iire reti-culoendothelial svstem anci
3-5

UNCIASSIFIED

IINCI-ASSIHED
generaLLzed distribution in the body associated with a more or less constanE bacteremia; and the preterminal septicenlic stage, which is

tracheobronchial nodes. During this time or after reaching the lymph nodes, the spore germinates and develops into the vegetative ce1l; infection begins by proliferation of vegetative bacilli freed from the phagocytic cel1. This vegetative ce11 moves along rhe efferent lymph channel through the lymph duct into the venous blood.(s'1s) There is g eneral agreement that B. anthracis characteristically does not develop vegetatively in the tn.tg to ""Glneumonia. (4) Apparently, organisms entering the blood from the ly*ph sysEem do noE proliferate in the blood, but are retsained by and multiply in the tissues of the reticuloendothelial system, chiefly the liver and spleen, until they exceed the retaining capacity of the tissues. Then vegetative ceLls pass into the bloodstream, and terminal septicemia is initiaEed. Multiplication of organisms conEinues until the death of the animal. The extent of growth of organisms, or terminal Ievel, depends on the reLative-immuniEy leve1 of che host. During the septicemic phase in guinea pigs (6 to t hours before death or 3 hours before the development of the critical blood bacilli concentration), 61 percent of the bacilli in the animal body were in the spleen and 16 percent were in the blood. At death, this ratio changed to 16 percent in the spleen and,72 percent in the b1ood.(s) After this critical 1eve1 of bacterial concentration is reached, the septicemia may be eliminated by antibioEic therapy, but the animal will die because of Eoxic reacLion. The time to death is closely relaEed to the time afEer reaching the critical concentration before initiation of antibiotic treatment.(e) Death results from the action of toxin(s) released by the growing organisms, which predomiaately produces symptoms of shock in the hosE. The implication that tosin alone accounts for the anthrax virulence has not been definitely confirmed.(5) Whether these experimenral observations in animals are duplicated in humans with anthrax is presently unknown.

the stage most characteristic of this disease.(s) Histological methods have shcrrn that spores deposited from an aerosol onlo the epithelial surface of the lung are ingested by macrophage ce11s on Ehe surface of the lung. These cel1s may be sessile or mobile. The mobile macrophage cel1s containing ingested sPores migrate through Lhe undamaged epithelium and enter the lymph stream, then move on to infiltrat.e the

(5) The extracellular toxin produced by B. anthracis contains three separate componencs: the edema faclor (EF), protective antigen (PA), and the leEhal factor (tF). These components are not toxic when injected alone into animals.(]5) The edema factor remains biologically active and produces an edema when mixed with protective anEigen; however, the letral factor is no longer lechal when injected in combination with proEecttve antigen. In monkeys, the ultinnte effect of the t,oxin on the central nervous system'is anoxia, that is, deprivation of oxygen to the body tlssues. This is caused by a lack of oxygenation of the blood, either ry itself or in combination with decreased blood flow. The

3-6

t]NCI.^ASSIFIED

terminal anoxia could arise through (1) a direct effect on the respiratory center in the central nervous system, causing a respiratory failure and a lack of oxygenation, (2) a teEanic paralysis of the intercosEal and diaphragmatic muscles, arising from increased cenEral nervous system (CNS) discharges and again resulting in respiratory paralysis and lack of oxygenation, and (3) a cardiovascular failure mediated by the increased cent,ral nervous system discharges, Producing a generalized smooth muscle constriction. This would also contribute Eo lack of oxygenation through bronchial- constriction.(17) The results obtained with the inacEivated forms of the toxin (inactivated wiCh antiserum or heat) suggest that survival depends on continuation of respiration, because the toxin comPonent affecting the respiratory center has been inactivated or because the components thaE cause the CNS-cardiovascular failure have been altered in iuch a \ray thaE rhe animal is able to reestablish enough blood flow to prevent terminal anoxia.(17t18) in" components(s) of the toxin specifica1ly responsible for the central nervous sysEem action have not yet been idenrified.
Almost all animals, including humans, f.. Susceptibilitv/Severitv. areto'o@b1etoanthrax.Theherbivoraarethemost susceptible; of these, the disease is most common in cattle and sheep' It is also common in horses,'mules, and s\^Iine, although swine are less susceptible. Humans are considered less susceptible than the herbivores' Carnivora may be infected, including domestic dogs and caEs, but Ehey are quite resistant to Ehe disease, part.icularly dogs. Birds are infrequently infected naturally and are infected with difficulty artifiRepCiles, amphibia, and fish may be infected if the body temPcially. erature is mainEained at a high 1eve1. Guinea pigs and mice are commonly used as experimental animals for anthrax and are highly susceptible. Less than l0 sPores of a virulenE strain of B. a+thfacis can cause death when injected into a mouse. ftaLs, particularly white rats, are very resistant. to anthrax.(tt4'6) The severity of the disease in animals is related to the natural susceptibility of the animal and the mode and 1evel of infecEion. Natural occurrence in animals results primarily from ingestion of contanrinaEed food. In endemic areas where food may easily become contaninated, large numbers of animals may be infected. AbouE 50 cases

of anihrax in humans are reporCed annually in the United States'(I'3'19) Excepc in rare cases, these are cutaneous anthrax' g. I"lortality/Sequel-ae. In the Uniced States, Ehe mortality rate reported .uTZiEiiiJi?Ections in humans prior to Ehe discovery of for antibl-otics was 20 Percent. Cutaneous anfhrax responds to treatment with antibiotics, and ihe moriality rate for promptly diagnosed and iratory treated cases is essentially zero. f"l!!!=-js--es-s-e+Eial-1:-1-g9-!::-.Jlc. antibiotics is inef f ective anEhrx is: uniikely, treiiment-;iih l--#
._
When

3-;

99.9 ooa oo <

99 98 95 90

+3

LD=

Based
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oa 1.235 animals

41130 spores, with 95% confidence of 1,980-8,630

Probit slope = 0.669 probits/1og dose, with 957" conf idencs lr'mi gs of 0. 520-0.818

+2

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70 60 50
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0)

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30 20 10
q

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0.5 0.2
102

103

104

l0s

Dose (spores)

Figure 3-1 (U).

Response of Cynomolgus Monkey to Aerosols of B. Anrhragis(ar) (U)

3- 10

:\

i. i_ .f y _y:-."\ ' *: -r

al at

c. (u) rarElcre ptze ano rnrecELvl-cY (1) As illustrated in Figure 3-2r(za) lhe ma-imrrm lrrng roEenrf nn 2 microns in diameter. in humans occurs r,rith the inhal@ affecte<i b n ' and hygroscopicity of inhaled particles. The curves of Figure 3-1 portray t@toti1upperrespiratory,anddeep-1ungdeposition versus particle size, at a breathing frequency of l-5 inhalations per rninute. The information in (a) through (e) be1ow is stnnmarized from these curves. (a) The highest probability for deposition of inhaled particles in the respiratory spaces of the lungs is in the size range from 1 to 2 microns (graviEy serrlemenE) and the submicroscopic size below 0.2 rnicron (precipitation by diffusion).

(b) Above L to 2 microns, penetration to and deposition in the lobules falLs off with increAsing size, because fewer particles escape upper respiratory trapping. Above 10 microns, the probability for penetration to the lobules is essentialLy zero. (c) BeIcru 1 to 2 microns, lobular deposition falls off, the efficiency of removaL by gravity settlement within the because lobules Ehemselves decreases. (d) The lowesE probability for deposition of inhaled particles in the respiratory system is at 0.25 to 0.50 micron, where the combined forces of precipitation by gravity and diffusion are at a minimum.
(e) The probabiLity of l-obular deposition increases as particle size goes dovn (in conErast to gravitational setElement, which decreases with sizel . (aa) (2) Monkeys often breath through their mouths resulting in varied and scattered particle retenEion in the lungs; aerosol particles 4 to 6 microns in diameter Ehat would have been extensively crapped in the upper respiratory tract e/ere Erapped in the lungs instead,.(7) If infection can only be caused by spores in particl-es with a critical diameter of less than 2 microns, present methods of assessment may be indicating incorrect dose patterns. I'or several years, field test and chamber work for dose response has employed the preinpinger-impinger combinat:on for dose determination of particles 5.0 microns or less. Assuming a 50-percent cutoff of 5.O-micron particles by the preimpinger and a mai:imum spore population for parcicles of given aercsols, then

3- 11

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DiameLer of Particle (microns)

Iigure 3-2

(U).

Total and Regional DePosition of Inhaled Particles Relation Eo Aerodlmamic Particle Size (u;.rzs1

3-12

partic:es with an OID- of 5.0 rnicrons nay theoretically contain as many as 75 spores per particle or as few as one. Cn the other hand, particles with ar OID of 1.5 mlcr:on contain only one spore per particle. Depending on rhe aerosol, this inciicates that when tl're cloud is assayed with the preinpinger-impinge:: combination in series, dose response for Particles 5.0 rricrons or less rnay vary for a given species by as much as 75, even :-f equal infecting capaciCy ls assumeC for inhaleci particles fron 1.0 to 5,C microns.(?)

I I

I I I I I I I I

I I I I I

44 .O

Storage

a. (U) General. Bacillus anthracis is one of the most resisrant of \\.,/ t'/ biological agents to deterioration in storage. Few data on the effect lythe //', of storage are available, because of the very long time required to observe t such effects. The viabiliEy half-1ife of the spores in storage has been " oC. i estimateC to exceed 5 years at temperatures ranging from 4 to 39

4-)

e. (U) Accelerated Assessment of Storage Stability (1) (U) Storage stability of both wet and dry B. anthracis were evaluated using an acceLerated storage concept.(43)' Thfr .."."pt useC a probit mccel to <ietermine the percent of agent recovery when the agent is stored at a constant temperature for given periods of time. In essence, the experi;aents consisted of placing agent sanrples) contained in glass tubes, in oil or ethylene-glycol baths (maintained at constant temperatures bei:seen 50 and 100 cC.) for 1 to 6.1 x lOa minutes. The data

/.4

I I I

t I I I

I I I I I I I I

(2) (U) Using the survival data at high temperatures and the model (Equation (4.3)), estimates of times to 50-percent survival were made for storage temperatures. The reported(43) estirnates of 50-percent survival are 620 years and 165 years for wet and dry B-anthracis, respectively, stored at 3 oC. Using the mode1, SO-percent survival time at roon temperature (27 aC.) was estimated to be 3 years and 4.5 years for rvet and dry B. anthracis, respectively. For storage at 39 oC., estirneted values were 12 r+eeks anC 43 weeks for wet and dry rnaterial, respectively. Although these estimated values indicate that B. anthracis spores can be stored. with relative ease for long perids, tn"y-ipp"l. Eunderestimate survival, when compared with the results of a few storage tests. For example, it has been reported that wet B. anthracis spores were stored in a laboratory for 5 years without detectable loss in spore viability, whereas the estimated value is a 5O-percent loss in 3 years. f. (U) Estimates of Storage StabiliEy . It appears to be feasible to obtain daca on loss of viabiiity of B. anthracis spores stored at room temperature (27 oc.) by actual measurement over a reasonably extendeci period (5 to 10 years). Such data would be of value in assessing che logistics burden for employing thi-s agenE cffe:rsively by an adversary. For the present, based on data in Table 4-3, it appears reasbnable to assune a half-life of at least 2 years for eiCher wet or dry agent storec Cn o^(,. dL !-/ It nav be assuned that l-oss of viabilitv of B anihra cis

dq-yonT e
ie

L-:

^7'

rr

I
,l

data are available on the effect of freeze-thaw moisture content of cvcling on dry B. anthrasis . Because rrf the smallof drYing, freezethe age:':.t and beiause it is i:ozen in the Process survival' thaw c'.'c1ing 1 ike l-v ha s lit iie or no e!fect on agent

(5)

(u) No

4-4.C

DisseminaLion

a. (U) General. The dissemination component of the source model describes the transformation of the biological agent (aqueous slurry or presized dry particles) into a viable, infective, inhalable aerosol c1olaofspecifiedstrengEh,size,and1ocation.@

ased on

pff_enE*wfEh-a

3-: L- a genL --t-q

:bgE s rlgr

b. (U) Source Strength

h" l:roma1it-g*rw".ry f."- " r.t"t-** -an instanLaneous diEseminaElon by an explosive dy!q-q-o-E a-continfTo..r -f .o* .i' ro qf ffi.t t"- tn. Ei: -ii-uT a f io.r1 uffii " "" "
(1)

siu?cE-stienfEtr oT-"t6e.aerosol Cloud - that is, the number of spores contained in Ehe cloud at its source - is a function of the quantity and concentration of the agent product in the disseminator and of the dissemination efficiency of the disseminator. IAerosol particles equal to or tess than 5 microns in diameter are considered optimum for e fficiency W e-E856fi-shing respira tory i":!igrr-irr-*3rr,-l The dis-s em

tG si"" range.l

(2> The various models used to determine the source strength a munition are defined as follows: of (") The source strength model for a point source of wet or dry agent material is
Q=
rvher e

VG.rd

(4.5)

.source Q=

with <5 microns ciiameter.

strength, number of viable organisms in particles in Ehe munition.

Che

V = volume of agenc fill

G.,

concentration cf viable organisms in ' = of muni:icn iurccioning.

agent fill

at

Cime

4-L4

d= efficiency of the munition in converting agent fill

into an particle cloud of viable sPores; the ratio of aerosolized che number of viable spores aerosolized in Part.Lc1g9 with <5 agent '@lE d_i_es"_!_.-'_ !_-o_ -q[9-_t.tr&9l_ s! _y=ie.hle- -fggls!_l! lhe fill prior to functioning .

q=-

Errd
(4
II

.6)

where q = number of viable organisms in aerosol particles with microns diameter, pr unit of line length
E = agent emission rate

<5

V = speed of disseminating delivery vehicle

(b) A line of relativell' closely spaced point sources (bcmbor generators) may be Ereated as a line source when the effects lets considered are at sufficient distance dorunwind for the individual point source clouds to merge.(4E) In Chis case, the source strength is
Number of point so"rces x a q=@

(4.7)

I I

This moiel is further discussed in Volume

X.

(3) Methods of measuring source strength are described fu1ly in Volume VIII, Part One. In some discussions, source strength is defined as the number of organi"E_Ig,q_9-9f thragent thE-t--;r,-e-b--ot-h--v'Lablc ement of the agent in an aerosol aF tu However, C1oud,--wEen'employed as a weaPon, is thaE it be infective. the source strength of TR has been measured and expressed only as viable spores, and Chis definition of source sErength is used in this part of the Source Book.

I I I

c. I

Dissemination Efficiencv

!r
I I

ffi

4-L5

CHAPTER

I
I I
. (U) Physical ProtecEion

orru*ru (u)

The protective mask is the principle means of physical protection from infection by an aerosol of TR. Eguation 7.1 is a general presenCation of the functions associaced with the penetration of the protective mask by a biological aerosoL:

I I

dr = rlr,B(t)

Ct

(7.1)

where d, rePresents Ehe respiratory dose (organisms)

r is a paraneter represenEing the fraction of inhaled

1

agent

I I

retained by the lungs a proEective

mask

r ls a parameter representing

the fraction of agent Penetrating

r = 1.0
a.

I I I I I I

B(t) is breathing rate (,/nin)

C is concentration of airborne agenE (o.gl !,)

t is exposure time (min). Volure X of the Source Book discusses chese respiratory urodels and paraneters in detail. As indicaEed in that volume, the reEentivity parameter is the only agenc-dependent parameter.
There are no data from which Eo estiloaEe Ehe retention of aerosolized TRI and TR2 in the human 1ung. However, data are available to indicate that 20 to 25 percenE of aerosol particles equal Eo 5 microns in dianeter

7-L

are retained in fhe human lungs after exPosure; a maximum of 45- to 50-percent reEention occurs with particles 1 Eo 2 microns in diameter.(as) 7-3. (U) Biologieal Countermeasures

Countermeasures agains E B. anthracis infection include chemotherapy, vaccinaEion, and supportive clinical procedures.

a. Chemotherapv (1) Effective chemotherapeuEic treatmenE of pulmonary anchrax in humans is possible if diagnosis of the disease is made early enough. If anthrax is suspected, creatmenE shouLd be initiated prompEly wiEhout waiting for laboratory diagnostic test results.(8r?o) Drugs must be administered during a definite stage in the disease before the onset of Eoxemia. Beyond this critical point, the sepLicemia wiLl resulE in toxin-induced deaEh, regardLess of anEimicrobial chemotherapy.( a rlo) (2) B. anthracis is susceptible to the action of penicillin and the tetracyclinesr- and these drugs have been effective in EreaEing cutaneous anEhrax.(3rru) Tetracyclines are less effective than penicillin. Table 7-1 presents the reconmrended drug doses for effeccive treatment of anthrax. The use of penicillin or the tetracyclines in humans as chemotherapeutic agents for preventing inhatation anthrax or for aborting anthrax infection following aerosol exposure has not been assessed. However, prompt treaEment of exposed individuals would presumably reduce infecEion and clinical disease raEes.
Antiserum

(1) Antibodies are effective against the causative organism of anthrax. They are not effecEive in neutralizing the metabolic products of B. anthracis, i.e., the toxin(s) which is produced in the host in large amounts during Ehe septicemic sEage of the disease.(s) (2) A specific antiserum must be empl-oyed to neutralize toxins associaced wifh anthrax. AnEiserum treatment prevented death in monkeys which had been given a normally fatal dose of anthrax toxin either intravenously or by aerosol. A1so, \"/hen treatmenc was initiaced afEer the septicemic stage of anthrax was reached, significanEly more animals survived with combination antibiotic and anEiserum treatment than rvith antibiotic Eherapy a1one.(5,'to ) AnEitoxin, in conjunction with bactericidal antibiotics such as penicillin, is recornmended. ( l 8) For anEhrax in tne septicemic stage, the dosage of antitoxin serum should be large "...40 ml intravenously, this amount being repeaEed several times a day. Urgerc cases have sometimes been given 100 co 300 ml inEramuscularly and inEravenously daily f or f ive da;rs. rr( " I ) Th. serum is nonstandard and is derived from inrnunized

7-2

TTNCLASSIFIED
horses. Severe serum reaction may occur, prolonging morbidity'(ztl "o) Antiserum is no longer available cosrnercially in the United SCates'(8 ,' appears thaE the Russians use antisera in the medical treafment of It anthrax. Also, the Russians use a varieEy of drugs that would be expected to stimulate the cardiovascular system'( I8)
Table 7-1 (U)
Drug
Reconrnended Chemotherapy Regimens

for AnEhra{3,1o t"o)

Length of nt Trea

Daily
G

Dose'

Adminis tra

Route of

Penicillin

PENICTIIIN U S trep Eomycinb Te tracyc 1 ine s

600,000 units 20,000,000 units

1 to 2 grams 2 grams

I I I I

tncramuscular Intravenous fntramuscular tntramuscular

7-14 days 24 hours 7 days 7 days

aBeginning at onset of disease (suspected or diagnosed). rstieptomycin and penicillin gdministered concomicantly have synergisiic effects on anEhrax disease of rhesus monkeys.(e) Such efficacy remains Eo be demonstraEed in humans'

c.

Supportive Clinical Procedures

(1) Experimental use of isoproterenol (1-(3,4-dihydroxyphenol) 2-isopropylarninoethanol) in rhesus monkeys resulted in Eheir surviving the effecls of anthrax toxin. Its action may be attributable to dilation of the pulmonary vasculature, which would a1low uninterrupted flow of blood through the vascular system of the lungs' or to Lhe known action of isoproterenol on the nryocardium, which causes an increased cardiac outpuc. One or both of these effects could maintain or the reestablish che function of the central nervous sysEem' especially The and oxygenation'( t") respiratory center, by increasing circulation effecEiveness of using isoproterenol against anEhrax in humans has not been reported Ehus far.
and fluid-eiectrolyle balance also requires special attention during the critical phase of anthrax disease '(25 '7o) This balance has been maintained in monkeys and found to be effective in preventing death. In addition' since the orygen level of che blood is decreased by toxins during the course of the disease, ox)'gen administration is probably helpful' Survi.ral of anrhrax-infected rhesus uronkeys was maintained by the use 5 ,1 o f a posici'/e-Pressure resoi rator'( ")

(2)Supportivetreatmentdirectedtowardmaintainingcirculation

12

U}{CTASSIFIED

UNCLASSIFTED
(3) Blood calcium and glucose levels decline before death in exPe rimen Ea1 animals. Therefore, adrninistration of calcium gluconate seems to be desirable. Steroids, administered after the bacteria that are present have been controlled by antibiotics or after symptoms of shock, also appear to be beneficial in reducing clinical symptomatic response to the disease. (6 )

d. Inrmunitv (1) It has been hypothesized that workers in factories handling conEaminated hides or wool develop a natural iurnuniLy to anthrax as a
contaminaEed environment. This has been presumed because the greatest number of cases of industrial anthrax have been observed in first-year employees. However, in evaluating the occurrence of anthrax among workers in contaminated industrial plants, Brackman, et a1.r('tz) concluded thaE vrorkers did not develop subclinical infection or inrnunity resulting from long exposure. They calculaced an atEack rate, based on incidence of disease and years of work, which rvas sl ightly buE not significancly greater fcr l- to 4-year employees than for 17- to 2O-year employees. The greater number of cases among shorE-Eime r.rorkers resulted from the significanEly greater number of employees in thaE category. demonstraEed by irmnunogenic response.('7o , '73)

result of a subclinical infection derived by exposure to a continuously

Irnmunity apparently is acguired with sublechal clinical

infecEion,

as

(2) Ilost of the recent inununological work in the United Scates related io B. anthracis has consisted of improvemenE of che protecEive antigen preparation for in'rnunizaEion of humans, clarification of che relaEionship between the proEective antigen and other components of the anthrax toxin, and investigation of the effects of innnuniEy on infection
and disease.(s)

(3) T\^ro human field studies of inhalaEion anthrax using Wright's vaccine nave been reported.("c,'71) Wrightrs vaccine is produced from the R1-N? strain of anthrax. RI-NP is a noncapsulaEed, nonproteolytic mutant oi the Vollum strain of B. anthracis. The protective antigen \ras precipicated and concentrated by adding 0.l-percent aluminum potassiu:r sulfate (alum) and was standardized against rabbits.(zs) The intracu:aneous LD.n of the preparation for rabbics r,/as approximately 100 spores.(-+) This vaccine was an effective inrnunizing agent for LaboraEc:y animals.(7s) It was well tolerated in a EesC in wirich approxi:.ate1)r 700 humans received more than ?,000 injections of the prepara:i6rr.(24)
One study involved r+orie:s engaged in manufacturing coaE from imporred goai hair conEaminated by B. anEhracis. Enpi-oyees'..rho had recovereci from anflirax \nere excluded froin :he Eesc.('o)

interli::ings

T]NCLASSIFTED
1t
l-;

a;ilrt-El,il8

TINCLASSIFIED
Two groups of volunceers were used. The same vaccination schedule was employed for both groups. One group received a placebo of 0.l-percenc a1um, and che other received Wrighc's vaccine. The iniEial series consisted of three 0.5-cubic-centimeter subcuEaneous injecEions, each given at intervals of 2 weeks; Ehese vrere followed by three booster doses of 0.5 cubic centimeter, which were given every 6 months; then 0.5 cubic cenCimeEer was given annually.(22) Forty-seven percent of a coEal eligible Eest population of 11249 people worked in high risk areas of a mill (spinning or carding of the raw wool), and 53 percent worked in
DaEa

low rlsk areas (weaving and finishing of interlining material).("o) were sEaEistically analyzed for inrnunized high- and low-risk groups. The attack rate in the placebo or conLrol group was calculated per 1r000 person-spnths. Based on this rate, the total expected cases of anthrax for Ehe tesE or vaccinated group lras estimated to be 13.35. Since only one case occurred, the vaccine appeared Eo be 92.5-percentseffeccive. The Lower 95-percent confidence limit for effectiveness of the vaccine was calculated to be 65 percent.(7o) The human experimentation results suggesLed that three consecuEive doses of the vaccine are reguired as part of the initial immunization regimen. One or trwo doses, administered 3 months after the primary series, seenred advisable for adequate and conEinued protection.( 7o

(5) A second iumunization program v/as conducced with human volunteers. Three subcuEaneous inoculations of 0.5 cubic cenEimeter of Wrighcrs anEigen were adminisEered at 2-week intervals. A booster dose of 0.25 cubic centimeter lras given 6 months after the initial series. In 660 individuals who received Ehese inrnunization regimes, no significant syscemic or 1ocal reactions were observed.(z+) However, no data are available relaEing to the degree of procection afforded these volunEeers by che vaccine against anthrax infection.(7a) (6) A number of live vaccines have been developed for anthrax, the first by Pasceur. I'{osE of these vaccines have been relatively reactogenic, and their use has been linited primarily to livestock. The Russians have developed teTo sErains of B. anthracis for use as live vaccines, the STI and the (Sh)-15 sErains, and Ehese have been used against l-ivestock.(5) The STI strain has also been used in the USSR for inmuniza!ion of humans by subcutaneous, dermal, and aerogenic methods.(75) Dry material was used in aerogenic inrnunization. Aerogenic inrnunization was reported to be least reactogenic and was as inununogenic as fhe subcucaneous injection and more irmnunogenic than the skin method.("5) Vaccinations are routinely given to individuals employed in tanneries and meat-packing plants or to people living in areas where anthrax is endemic. Vaccination with ST1 is reconunended at intervals oi 10 months.(zz
( 7) AnEhrax irununi ty is nore cotrplex chan other irnrnunities where serurn antibody tlter leve1 is considered a neasure of the level of i;rrnu:::Ey. The naEure of the anthrax antigens and the stage of the

7-5

t]NCIASSIFIED

-T
1
I

I}NCLASSIFTED
disease affecced by a particular antigen musc be considered.(za) To provide maximal inrmunity against anthrax infection, antibodies against spores) vegetative ce11s, and toxin may be required. The immunogenicity of components of anthrax toxin has been studied. These components have been described as the edemic factor (EF), protective antigen (PA), and Ehe lethal factor (LF); as nentioned previously (par. 3-3e(5)), they are noE individually toxic, Studies have shown thaE the LI component was irrnunogenic for the rat and the guinea pig against spore challenge and for the rat against toxic challenge.(?erao) (Although the raE has low susceptibility to infection, it is highly susceptible to toxin injection.) Inurunity of the rat to spore challenge was increased when PA was included with LF in inmunization. Inrnunity to Eoxin challenge did not irnprove. PA, as weLl as LF, inrnunized the guinea pig against spore challenge, and the conponents vrere complimentary. The variation in results between animals enphasizes the problem of developing effective human anthrax vaccines derived from toxin antigens. However, it appears that the LF and PA cornponents eould now be developed as a vaccine against anthrax in humans. Another problem raised by inrnunization, particularly against the spores or vegetative cells, is the potential for 1ow-leve1 infection Ehat would now be detectabLe through observation of bl-ood sanples, but would al1ow accumulation of toxin to a critical 1evel. There is no basis to predict whether or not this phenomenon should be observed in humans.(eo)

tt ',
d

,i

!
'!

T
i

7-4. I
a.

o""oniamination

(U) Di:;[er]!.rces in {gqnt

Forms

(i) VegeEative ce1ls of anthrax are to some degree resistant to deleterious effects of environment; however, in Che spore form, thi organism is verl resistant to environmenEal effects.(3 rs) Spores rqi11 * not germinate to form infecCious vegeCatiwe bacte.4ial--Cells- u,qJg_s-_s exposed to air for several hours at 68 oi_r_(ZQ_"gJ___at a relativg humidity equal co or greaEer than 65 percent. (2) To insure effective decontamination, deactivation of che spores is reguired. A summary of the resistance of B. anchracis to environmental or other decontaminating agents is as follows:(3,1e)
(a)

Vegetative cells: Are ki11ed in slurry by heating aE 65 oC fot 30 minutes Are readily destroyed by disinfectanEs, such as 1ysol, betapropiolactone, or aliohol A:e kilied by heaeing aE 55 cC. for t hour, or several mirutes a. temperatures above 70 "C,
7-6

I,INCIS,SSIFIED

(b)

Spores:

Generally are ki1led by boiling for 1O minutes Generally are killed by exposure to dry heat at 140 oC. for 3 hours
Are kil1ed by exposure to vapor of betapropiolacEone or formaldehyde for 2 and 16 hours, respectively, at a Eeuperature above 70 oF. and a relative humidity above 70 percent

Are resistanc to drying effects of environmenc and can remain viable for 10 years or nrore
Disappear rapidll' from unprotected soil surface exposed to direct sunlight for a 2-day period
&!

when

;I
I I I

Survive for long periods under proCection of soils, in animals hided and carcasses, and following aero so 1iz ation
Are able to survive for long periods at cryogenic terperatures (-120 oC.); are also resistant co rapid freezing (-78 oC.; and thawing (37 oC.) procedures.

(3) Irradiation of spores dried on glass (e.g., window glass) trith ultraviolet at an 18- co 20-vo1t intensity (simulates a clear day) for 45 minutes produced an 89-percent reduction in spore viability; irradiation at a'16- to 2l-volc inEensicy (natural sunlighc) for 45 minutes reduced spore viabilit;l by 67 percent; with a 60-minute exposure, the reduction was 95 percenE. Vegetative cells can withstand short exposure to ultraviolet (75 sec.).(ee)

I I I

I I I I

7-7

(2) (Lt) ZLrLa, etal.,(Bs) in an analysis conducted for the U.S. Air Force, iodicate that betapropiolactone (BpL) is effective in decontaminating surfaces conLaminated with spores of B. subtilis and chat it is the least corrosive area decontaminant pre"EnEty-Z"aTlable. Requirements for speciaLLzed decontamination of equipment and surfaces contaminated with microorganisms or toxins led to the investigation of formaldehyde gas released from dry paraformaldehyde as a decontaminanL.(e6) The sterili-zi-ag effectiveness of fcrmaldehyde gas produced by depolymerization of dry paraformaldehyde i.s given in Table 7-3. Values presented for B. subcilis apply to spores of that organism and are applicable to spores of B. anthracis, inasmuch as the spores of B. anthracis are less resistant tnan spores of B. subtilis. SterillTarion occurred after depolymerization of 0.3 gram .f p"r"f.r-aldehyde per cubic foot o: space for a l-hour contact period. The range of use for dry paraformal-dehyde is rated equal to that of formalin. Test results indicate that the dry gas produceC by depolymerizaEion of paraformaldehyde may be a more effective sterilizing agent than vaporLzed formalin. This gas also disseminates irore readill'and is more rapidly dissipated by aeration. Formalin (37-percent forrnaldehyde in water) was rated equally as effective as BPL, but the requirement of a longer contact time for the formalin nakes it less desirabl"e. (85) Ethylene oxide vapor is somewhac less effective than either BPL or formalin, but it is a useful decontaminant under certain circumstances. However, ethvlene oxide was one of the mosE corrosive of the decontami-nants tested on Air Force materiel.(Es) Table 7-4 contaios some pertinent information relating to the cited decontaminanCs.

;- t0

rI

lG3:r-t:i$**t.:_

*qd&*|l:btd.

Itul,l!tu'.-r@E.

Table 7-3
TesI

(U),

Formaldehyde Gas
Vo lume

SLerillzatlon of Facilities, MaCerials, and Equlpment

Anount of
Paraforrnaldehyde
(em)
Conc ent ra t

(lrlnrli t i ou

(crrblc fecL)
2250 4s9B
67 2L6

Organl

srns

(per m/)
106 1010 106 106

lon

Viab I e

Recoverlesa

T.,aboratory rooms
La

685

subtilis
marces cens

L379

0/s 0/s
0/L

rge

roorn

20t65
330

srrbtllis
sr-rbtili-s

{
I

Iloblle laboratory
Surfaces Iri l

2200
25

0/s
0/s 0/s
0/ 10

H H

7.s
12,6

subEllis

10?
b106

rcr rncdia in class I cabincL

c

42

subtills
B. subtilis B. subcills S. narcescens

Laboratory
<lul pmen

100- 200

30- 150

106

Vacclne tubcs

0.6

104 104

0/2 0/2

"Nunrber

bCoucentration of 106 per paLch.

of viable recoverles per total tests conducted

n 'i
;r

I :!
{
!:

.t'rble 7 -/+

(1:)

Characteristics of Selected Decontaminants(85) Physical State for Use

Vapor or aerosol
Decontami nant Requirements

I'

Decontaminant

Environmental Limi t at ions

RH

Betapropiolacione
( nPi,)

2-hour contact time;24-hour aerat ion

not less than

7O%: minimum

effect ive
temperature 24 oC. (75 oF.)
$

Formalin

(37%

formaldehyde in wat er)

Vapor or
aero so 1

time;24-hour
aerat io
n

16-hour contact

RH 857. opEimum;

than l-6 oC. (60 oF.))

Ethylene oxid.e
Vapor

Eemperature 2l27 oC. (70-S0 "tr'. ) (not less

6-8 hour contact time at 2L ac. (70 or.); 12-hour contact time at 16 oC. (60 or. ); 12-hour aeraEion

Minimum

effective

16 cC. (60 oF.)

temperature

-L2

I I I I

I I I I I I I I
\

(5) (U) Findings coocerning the effectiveness of BPL in reducing or eliminating the contamination wilhin the ship are noE included in the cited report.(87) The data'wera far too variable to give explicit results; horvever, the data showed that contamination was naterially
decrea s ed
.

(6) (U) fhough BPL was found to be an effective and relatively non-corrosive decontaminant, ils use as a decontaminAnL has been negated by a ruiing of rhe Public lleal-th Service prohibiting Ehe transport of BpL in interstaEe cortrnerce. This order was instituted because of the toxiciE)' and report,ed carcinogenicity of the compound. e. (U) Decontarnination of Small Articles. Because of the inability of BPL to extensively penetraEe hard-to-reach.spaces, convoluted materials cannot be decontaminated using Ehis agent. ( u- I Ethylene oxide can be used to decont,aminate srnall ieems (books, too1s, etc.) by placing the contarninated articles in sealed plastic bags and releasing ethylene oxide (usually in an aerosol can) within the bag.(ee) As indicaEed in Table 7-4, 6 to LZ hours of contact, dbpending on temperature, should adequat.ell' eliminate Ehe hazard. Echylene oxide has beeq used Eo sCerilize laboratory equipmenE or cloEhing using Ehe gas in an autoclave, a sEeel drum, or polyeEhylene bags(48'8e) MaEerials heavily contaminated with resistaot spores will require an exposure of 300 to 500 milligrams of eth;rlene oxide per liter of air for about 6 hours at 25 aC. to destroy che bacil1i. Some moisture must be present to adequaEely compleEe the process of decontamination.(3e) No changes io the external appearance

of the articles (mi1itary uniforms, shoes, etc.) were noted after exposure of 4 to 10 hours at a concentration of 20 to 30 grarns of ethylene oxide per kilogram. B. anthracis spores were inoculated into cloth treated with various chernicals, including permaseptic and chlorine in an assort.menE of solvents. delorine in acetone tetrachloride was quice effective in rendering Ehe cloth unsuitable for organism survival. In aooLher experiment, boiling for 15 minutes rid the cloth of spores.(68) Soaking contaminated clothing in a 10-percent formaldehyde solution is effective.(6) Autoclaving at LzL oC. for 15 minuLes is a reliable treatment for kiLling spores on cloth or other small articles, but the efficiency of this method depends on the complete heat penetration of bulky material.(s) Polyadov, eE a1.,("o) suggested the use of nitrogen dioxide as a bactericide against both the vegetative and spore forms of B. anthracis. The high mobility, penetrating po!/er, absorptivity, and 6Ch-:contact properties of gas molecules gives- gaseous bacteriocides an advantage over liquid and powder disinfectants. Nitrogen dioxide (NOe) is a powerful oxidizer. Tests have shor'rn that NO, at a concentration of 0.5 gram per liter of air killed the vegetaLive forrq of B. anthracis in2Eo3minutesandsporeformsin5to].Ominutes.rt'esGffi made at room Eemperature and normal atmospheric pressure. Nitrogen dioxide should be tested as a disinfectant of different objects in enclosed premises, as well as of buildings infected with anthrax sporer.('o)

7-L4