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Project No. 5-C0-473-000-016 DTC Project No. DTC 7l-504

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L'OINT CB TECHNICAL DATA s ouRc E BooK tul
VOLUME VI I
I

Bacterial Diseases

(U)

*ln
l/..

Part Three: Brucellosis (U)
"
MAY I 97I

'rr f\:

I

rl.

'{

t.

\l

This raterial ccrtains infornation affecting :he national ce:-ense of thc United States uithin the mean:.ng of the Espionage Laws (18 U. S. C. ;9-i, 794) , the transmissicn or revalation of rrich in any nanner io an unauthori:ed :e:son is prohibited bv la*.

Drstrib'tion d^rrro to u. s . ffi^"^, (Test and Evaluation). agencies only. Other requests for this Vav l97l Corunanddoculent must be referred to: ing Ceneral , neseret Test Center, Fort Douglas, Utah 811 13.

b

L'-/

l
$ f\ i.
\

HEADQUARTER,S

.

DESERET TEST

CENTER

'

FORT DOUGLAS,

UTAH

.

84I I3

DTC 7l-3:.C
COPY

oF 172

COPI:

tINCI..A,SSIFIED
FOREI^JORD

This documenE was prepared in compliance with Depar::E:-: letter, "JoinE Contact Point for Chemical-Biologica- lI Data," 10 March 1967, which directed DesereE Test C::::: and mainEain a joint CB technical daEa source book.
The Source Book is organized into a series of volurrs addresses an identifiabLe area of informaEion relat=: of CB weapons and defensive sysEems. Areas include : models, weapons systems, assay and data reduction p:: simulants and biologicaL non-paEhogens, and knowlec:=

--::,:::::=' =:: :'-,::: which identify and define the parameters for which -;.::e:--::- .=-:=: =r= required in estimating capabilities of weapons syst=rs :: ::= -:-:-:: Services. Weapons systems which have been type class--=:=: :: =:= -: an advanced stage of devel-oprnenL have been included.
ParameEer values with confidence leve1s derived frc::::-and chamber test data are presented. Models and su:=:,:=-=

The Source Book is designed to be used by che resea::: ::.::: ment community as input into design and analysis o5 r.-.=::=: : def ensive techniques, and defensive devices. It n'€- : -- : :. those responsible for preparation of system perforn:::: :=:l: inclusion in field manuals, firing tables, and othe: ::::=::: munitions expenditure and effectiveness information.

Portions of the information contained in Ehe Source l.:,:. ---::: ::-= --:-by GEOMET, Inc. , under contract DAAD-09-69-C-0078, ;::-- :-: :::-j -::::: by personnel of the Joint Contact Point Division oj l:.::=: l:=: I.=rl-:l ::-=-:=All material has been subjected to review and coorc-:-::-:::members of the CB communiEy. The conscientious ef::::= :=:'=:::: :' : :::':-:: these individuals to improve the quality of the fie:---:gratefully acknowledged and appreciated.
Each part ai:d volume of the Source Book will be upc: --:'= ==:'Frequency of update will be dependent upon the leve- -- j ::: -the researc:l and testing areas covered by the appi::=:i: ::: volume.

regarding the adequacy o: material presented in Ehis document and any reques: the use of ;he documenE should be addressed Eo:
Corffnencs and suggesEions
Commanding General

Deseret Tesc Center ATTN: STEPD-TT-JP-I(S) Fort Douglas, Utah 84113

UNCI-ASSIFIED

SNou'\SS\sW
F

ILLUSTMTIOT*S

ierrre

Title
Stored

Paqe

4-l i'2 5-t 6-l

\X Vtablllty

Decay when

at 4 o to 6 oC.
Dcnrn-

4-a 4-10

Tl,e Relatlonshlp Eetween Hunltlon Spaclng and wtnd Dtstance to Cloud Overlap . .
trX

Torold . .

btologtcel Dgcay Rlte as a Functlon of Relaclve Hurnldlty 8t 21 oC.' (ZO o9.) Measrrred ln the Brlctsh
5-7

Releases Durlng Wlnd Speeds of 9 Meters per Second w!.th a Decay Rare of 1.5 Percent per Mlnute. .
Donnwlnd Dosage Predlcclons for Aeriel Releases Durlng l{tnd Speeds of 3 Meters per Second wlch e Decay rl,ate of 1.5 Percent per Mlnute. .

!- suls

Dormwlnd Dosage

Prediettons for Aertal

6-2

6-2 B. suls 6-3

6-3

B. suts Dorrrrwlnd Dosage Predlctlons for Aerlal Releasee Durlng tltnd Speeds of 9 Feters per Second with a Decay Rate of 3 Percent per Mlnute. .
Donnwlnd Dosege Predlctlons for Aerlal Releaaes Durlng Wlnd Speeds of 3 Meters per Second with a Decay Rate of 3 Percent. per Mlnute. .

6-4

6-4 B. suis

6-5

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flrlF*TABLES Table

UruO[ASSfFilECI
Page

3-l
?-t

Symptoms

of

Hunran

Tltle Brucellosls.

.

.

3-3 3-5

1949

Inctdence of Brucellosls tn Occupsttonal Groups for ln Mlnnesota, tZ\ and l,tlsconsln . . .
,/t

4-1

1-2
4-3 4-4 4-5 4-6 4-7 4-8

Storage Stablllty of NX ar -50 -C. . . . . Viabl1lty Decay Paramecer Values for Agent }IX 1n Storrge.....4-s

storage Stabttlry lx/.. 4 o ro 6 oc. "f \-/ o

4-3
4-5

Effect of Storage for 10 anl 60 Days on t{X Dissentnatl,o'n Paraneters. . . . .
Estfuoated

4-l

Efflclency for Munltlone Teered wlrh Agenr NX 4-ll

Esllnated Paraneter Values for Standardlzed Blologlcal Munltlons for Use wlrh NX.
Dlsseolnatlon EfflctencLes for M(

4-ll
4-13 4-L4
5-3

FortDetrtck.

......

Chaurber

Trlals at

Suooary of Test Flxture Dlsseolnatlon Efflctenctes for }tX Chanber Trlals at Fort Detrtck. . .

5-l
5-2

Sumary of Decay Data fro@ Fort Detrlck Chaober Tesrs
Conducted

wlth l0(. .

Toroid Test Chasber.

Blologlcal Decay of l{X as Measured in the Britlsh

.

5-6 5-8 5-9

5-3

B. suls Half-Ltfe as a Functlon of Dtfferent Relarlve Hnoldlty and Tenperature Cooblnatlons.

5-4 Calculated Blological Decay Cqastants for Collaborattve Tests 4, 4A, and 48 . . . . . 6-1 Esclnated Dc'ynwlnd Dtstance of E\o Produclng Dosages for B. suLs Releesed froo an Aerlal Llne Source. . . . 7-t Recmrended Chemotherapy for 3rucelle sutc Infectlons. 7-2 A-l ' of Antlblotlc Toxlc Reactlon . .
Fteld Trlals Conducted at
xL

O-l
7-3

Reconoended

Corticosterotd Therapy for the Treatnent
DugLray

l-3
A-l

Proving Ground

UT{CLASSIFIET)
fr

CHAPTER 1

I
t-t.OGeneral

r**"

(u)

l-2. OAgent

and Disease CharacterisEics

b. (u) other strains. Brucella Eli!ens_}t_ and Brucella abortus, Ewo othe: species of the genus Brucella also infective for humans. c. (U) Incubation Period. Variable; 8 to 30 days, but may extend to several- months. (Includes all the Brucella.)
I

d. (U) Duration of Illness. (Inciudes all rhe Brucella.)

Variable; averages 3 months'

I

I
il

e. (U) Morbidity Rate of Naturallv Exposed. Populations . From I to a0 tercent. (Includes al1 the Brucella.) f . (u) l'lqrtalitv Rate. An average of 2 percenl for untreated patients vrith B. suis infecLions. pa t ient s

F

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a. (U) Prcduction Slurry Count. 3:'.Ldo organisns per millir l-1 'f
-!
r
r

UHCLASSIFIED

b. (u) c. (u)

Storase Half-Life.

150 to 169 days

at 4 o ro 6 oC.

Dissemination Eff iciency.

T

(l) A/B45Y-1 and Aero 14B aerial dissemination tanks. Estimated Eo be 3 percent. (2)
M143

bomblet. Estimated co be 0.7 percent.

t-5. (U) Infectivity
Unknor.m.

Decay

t-U.

t a. (U) Casualty Rates. Insufficient data to predict.

Effectiveness Predictions

(U) Defense Against the BruceIla

a. Detection. No adequate detect,or available. b. Protection. Several adequate protect,ive masks available, such as the M17A1 and Mark 5. c. Vaccination. No reliable vaccine availaole. c. Therapy. Tetracl-c1ine administered join:ly wirh streptomyc::t is effective in reducing severity and duration of illness. e. Decontamination. Beta propiolactone, etirelyene oxide and o:rer bactericidal chemicals are effective.
L-2

I'tl{ {-

uiIt-LA))rlu
CHAPTER 3

I o.u*t AND DTsEASE cHARAcrERrsrrcs
3-1. l)
General Properties -

(u)

3-2. (U) Biological Nature of the Organism
The three species of Brucella are characterized in Bergeyrs ysnsal(2) on the basis of antigenic properLies, tolerance to a variety of dyes rvhen gror!,n on sulici media, and various ocher determinative techniques. B. melitensis was the first organiqm identified as the causaLive agent of brucellosis and was shoqrn t,o be transmitEed from infected goats to man through ingestion of milk. Subsequently, B. abortus rpas isolated from cattle, and B. suis from swine. These species have been found in many dcrnestic and semid.oinestic animals and in man. Some strains have been found. that are not readily assigned to any of the three designated species. Among the many strains obtained fro-r:r various animals and collecEed. in d,iverse geographic locations' a range of cultural and pachogenic characteristics have been demonstraEed within the confines of the d.eterminaLive criteria of the genus.(e) The Brucella are gram negaEive, nonsporulaling and nohmoCile. They are aerobic. However, soine sErains, particularly of B. alggg5r require an aEnosphere containing approximately 10 Percent carbon dioxide in order to grow in culture media after initial isolation from animal Eissue. The Brucella may be grown on a variety of culEure media. Cul Eure is nor affllgg}., al tho-u,gl-erow !h- _5_11s_t4!1y.- 39!-PI-9Ig!-a . *---.--.-^L1()^ --may ' ure is 37 "9_. _a19 rec9gnr14!_lg_c=*tg -^.. be ] "C. and recognizabl ^---rL

I I I I

I
I

3-3. D Strain Selection

and Developrnent(a)

I I I I

3-1

I i<f".

fr)

TJilCLASSIFIED

..

i.

3-4. (U) CharacEeristics of the Disease a, Nornenclature. The disease produced in man by the Brucella is brucellosis. Synonyms used to designate the dj-sease are Ilalta fever, undulanE fever, Ilediterranean gastric remittent fever, Neopolitan disease, Texas fever, Rio Grande fever, and Bang's disease. The disease in animals is known as brucellosis, Bang's disease, infectious aborE.ion, and epizootic abortion. . Transr,ris s ion. Bruce I Los is is character ized b1z extens ive invasions o' tissues of Ehe body by the infective organism. The organisms remain in the tissues for long periods. In animals, where the infection becomes latent, it may persisE for years. The Brucella are passed from rnfected animals, in milk, and in feces, urinE-ZnE-6Eher bodv excreta. The disease in man is most frequently found as an occupationally related disease of farmers, neat handlers, veterinarians, and others ergaged in handling aninals or in processing animal products.\! ':' c . S!-::.p t.oms
b

I I
I

I

j

(i) lhe onset of brucellosi-s uray be abrupt, manifested by chills, fever, and sweati-ng, ,or it may be an insidious development. manifesled b.7 weai<ness and mild incapaciEation over an extended period. Table 3-l- fists sl.rnptorns chai are predorninant and the frequency with which t:lel/ may cccur . The nani f es taEions and assoc ia teii severity of s1'rnptoms i,-,-e been -:sed to characte::ize f lve distieeuishacie types of the l:-sease i:. l'tumans :(: )
(.3) InterrlittenL :\'De.
Assoc:-aced witi
J-J

siriiting articular

UNUIJ\SbIf IET/
Table 3-1 (U). Symptoms of
Human

Brucellosira(z) Percent of otal
B

Number

Fever

L7L
140
L37

Chil1s Malaise
Weakness Body ache

9.5 73.3 7L.7

L28
128 128
99 75 70

Sweating
Headache

67 .O 67 .0 67 .O

Anorexia Weight loss

51.8 39.3 36.6

aProvisional data from 191 case reports in 1968.
rireumaEisi,r, !.'eakncss, ncc:ur::al p.3:spiration, anC a_ temperature near normal in the morning, but rising to 101 " to 104 eF. in the evening.

(b) Ambulatory tvpe. Exhibits the s€rme general the int.ermittent type, but symptoms are less severe.

sympcoms as

(c) Undulant type. Characterized by daily fluctuations in the body temperature generally caused by B. melitensis. (d) Malisnant tvpe. Sustained high temperature becoming prior to death. Almost always fatal, generally caused by melitensis infection.
exEreme
B.

(e) Atvpical chronic type. Manifested clinically by muscular sEiffness, gastric disturbances, and various neurological abnormalities. (2) BronchiEig may be associated with infection of the organisns through inhalation. ?ulmonary invasio+ may also be associated with infection t.hrough other modes of entry wherein the lungs are invaded as a consequence of proliferation of bacteria within the body. The nature of Ehe sJnnptoms for any given case of the disease canno! be aligned with a specific mode of entry by the causative agent,. The mos: distinctive syrnptoms are fever, frequently reaching 105 oF. and usual--;; fluctuaEing discernably, and enlargement of the lymph nodes, t,he splee:, and the liver. Mental depression also is often associated with the disease.lt r= re) d. Onset and Duration (i) Variation in onset, or incubaEion time, and in length of

\

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i
ITNCI-ASSIFIED

,,.-.,r.

=.sl*iGtn{raEH

UNCI.^H,SSIFIED
illness has no apparent relafionship to moi.e of infeccion or to Che strain of Brucella involved, as indicated by the variability observed in the outbreaks of the disease. The incubation period of the disease ranges from a few days to several monEhs, with an average of 8-10 days. In the case of laboratory accidents with B. suis and B. melitensis, the majority of cases have had an incubat.ion period between 4 and
B weeks.(z)

(2) Studies of brucellosis case histories have indicated a duration of debilitation serious enough to require hospitalization ranging from 20 days to over a year. LaboraEory acquired infections showed the s€rme general variability in duration to those by 'hatural'r means. When the disease is established as a chronic infection, Ehe debilitation may recur intermittently over a period of years.(s) The average incapacit.ation period has been estimated to be 3 months.(e)

Susceptibilitv and Severitv
l
'

g
'I

1
I,
I t i

(1) S:udies of groups exposed to the Brucella have indicated a rather wide range in level of suscePtibility of man. Tirese s E':3 ies , for Lhe most part, involved those'ocoupational groups most likely to be extensively exposed to the organisms. In one grouP of 120 veEerinarians, where Lhe probability of frequent exposure to infectious material exceeded 90 percent, only three indicated any clinical hisEory of brucellosis, although 45 percent had agglutinins for the Brucella' One of the group was exposed Ehrough ingestion and skin abrasion, but did not dev-lop clinical symptoms of the disease or agglut,inins for the Brucella. In another Sroup of 49 vet.erinarians, 75 percenE had for Ehe Brucella and Ehree had a history indicating .ggfffiitr" symptoms.(s'6) clinical

I'

I

(2) Incidence of brucellosis in occupational groups in four Midwestern States is shoqrn in Table 3-2. No precise assunption as to exposure rate for these groups can be rnade. However, it is believed that the rate
was high.(s)

T

(3) In sunrnarizing these data, it was concluded that in a population where the probability of exposure to Brucella is high, the yearly incidence of clinical brucellosis will not exceed 5 percent of the population, and would more likely be approximately 1 percent.(5) Other case studies citing specific Brucella species have indicated much higher susceptibility. (") Susceptibility of B. melitensls has been reported as high as 40 percent. (4) In an accidental exposure of students to what was assumed to be an aerosol of B. melieensis, it was reconstructed that a certain group of individuals was exposed to a high dosage, and anolher to a lower dosage. In the high dosage group, 25 percent were clinically ill, and an additional 30 percent indicatec no symptoms, but did dcvelop a
?-.

IINCIS.SSIFIED

UilCLASSIFIED
pos

percenE were

itsive irrrnunological response.

posiE,ive irrrnunotogical response. (s )

clinically i1l

and

In the low dosage group, only 4.5 an addiEional 11.4 percent shoraed

Table 3-2 (U). Incidence of Brucellosis in Occupational Groups for L949 Ln Minnesota, Iowa, and. wisconsin(6 )
Packing House
S

Eate

VeLerinarians Total I Total Pooulation I er.o=
740 403 502
9

Workers

Total Population
22,9L0 L5,235 8.497
a(),642

Total
Cases

Farmers and Ani:nal Husbandrv Workers

Total Populatior
265,500 249,700 267 ,300
782 ,500

Total
L73 L4L

Iowa Minnesota
Wiscons
TOT-^.L

in

4
2 15

4L 50 26

l1s
429

L,645
(%)

il7
o.25

Rate

0.9r

0.05

\

(5) Severity of illness varies markedly. Many cases are declared subclinical with the patient only slightly debilitated. The duration of incapacitation may be long, but in many cases illness would not be so int,ense as to preclude performance of necessary tasks. To some extent, severity of illness is related to the species of Brucella, with greates! severity resulting fron infection with E. melitensis. Severity and duration of the disease can be markedly reduced by treatment with antibiotics.(s '?)
(6) Variation in susceptibility and, severity of the disease with aB€r sex, and other population categories, has been found to occur to some degree. The young are more resisEant to brucellosis than are older people. The disease has seldom been reported in children under 12 years of age except in undernourished populations. In such populations, chfldren who contracE the disease respond as severly as do adults.( z )
f. Mortalitv and Seguelae

\

tI

(1) Mortality rate calculations for brucellosis are based on a fairly small number of clinical cases. The mortality rate determined for these cases varied frorn zero co 21 percenE. The highest mortality rate was reported for an outbreak €Lmong elderly persons. The besE esEi$ate for n'.orcalicy among a healEhy population with an al'erage age distribut,ion r..'ould be approxinately 2 perceng.(s)

1iry

c!, n 5.!rFIi:J

UT'ICLASSIFIED
(2) serious cornplications which m"y be persist,ent occasionbrucellosis. Recurrent chills and, fever, localizaEion or organisms in calcified necrotic lesions, episodic myalgia, granulomatous reactions appearing as rElsses in the spleen and liver are sc'me of the permanenE sequelae. Central nervous system effects, such as depression, frustration, anxiety, irritability, restlessness, an6 apathy, are often noted. organic lesions of the cerebral vessels, meningo-encephalitis or meningitls may occur. other organs which may be affected are the heart, (bacterial endocarditis), deep veins (granllomaEous changes in vessel walls causing thrombosis), lungs (pulmonary erobol-ism), reproductive systern (epididyuro-orchitis occurrLng in 2 to 5 percent of adult males), kidney, ureter and bladder (granulomatous lesions, sonetirnes calcification), and the skin (rashes). The conjunctiva, cornea, iris, choroid, retina,.opt,ic nerve, and. ext,ernal muscles of the eye also may be involved,.(6r7)
all-y
acconrpany

(S) InfecEivity

*^^.,,i:.I9l"if|iGTn estirnated to be 60
and the
)
t\

infectivity of B. suis

"rbffi"r"o.r

ED"o for

has been studied in for disease oduc t!s!_-!gr suinea. pigs,-lras with e dogs_re9l9lr,s_9 _cq11v-e---q lo-Le_igr = less t]ran 400 or_sarljsms

**k.

-es_g_-lha!

!pg_o:gar1!ggrs.(Ir)

)
t
i
t-

I
f;

c. (U) The Fort Detrick hu-'n EDso eslirnals was challenged in 1958 in an Operations Research Group study-.Is) thts study qualltatively analyzed data regarding an epidemic of brucellosis at Michigan State unLverslty durLng L939-4a. These data provided the basls for the comrqent that Ehe probable dose of B. melitensis required to cause lnfection Ln mrn by aerosol ray have been greater than 106 viable organisms per person. Hcnrever, no correlation could be made between the dosage to which indlviduals roay have been exposed and the percengage of Perssns who developed clinical synptoas of the disease.(E)

\;
I

3-6

F
p c!-*^

55lf

lli)

UHCLASSIFIE.D

fr
I

CHAPTER 4 SOURCE PARAIETERS AND MODELS

(u)

Agent Production

I
I
I

I

I
I I I

I I
S

(1) (U) Holding temperatures. (2) (U) Ntunber of freeze-thaw cycles.
Lorage contac t
L V.

conlainer material (especially the surfaces and coatings in rvitr the agenL slurry) may also influence viability. Effect of StoraEe on Viabilitv I (l) (U) For agent NX the reduction of viable cell concentration at constant ternperature can be described by the exponen:ia1
deca,'r nodel
:

I

I

c t-nroJ
e

a'-o-2oa
ev4

*Ce

!

I
.,\..!)iFi il)

c(t)
where

= co exp

(-r"t)

(4.1)

c(t)
-o

=

the viable ce11 concentraEion in cells per milliliEer a: time, t, t::e initial viable cell concentration in cells per rn:lliliter at the time of production (t = o)

(^= exponential viabiliLy decay constant related to s i.orage tirne , pe r da y t = length of storage time, per
The point in tine at which the viable cel1 concentration, G(t), in the stored material- has {ecreased to one-half the initial viable agent concentration, Gc, is the storage half-tife, Tso. By settinC c(t) - Go/2 aad using equation (4.1) Tso= o'd93
K
S

(4.2)

:
I
a-

t

I

I

I
i

(3) (u) rue data in Table 4-l were plotred as 1og percenr survival versus storage age, and were fitted with a least squares regression line (Figure 4-1). The slope, 0.0041, is rhe decay consrant. Using the stardard error of estimate, the 95 percent confid.ence limits , of the mean values of \ were deternined. The analysis of the data tn I presented in Table 4-1 gave a mean decay oi i,f 95 _percent ccrf idence 'lTrTEF-o.r--03f-to 0:15 percenr-peC day; the T^^ / | of EEE-sToieE miE;;ia-l- ;as'?pproxirnarely 169 iays compared. wirh rhe previously re2orred half-life of t50 days.(13) This longer half-life nay be due i:: part to the storage of the agent in glass containers. -aLf-1:-fe r-r4s re,ported.for agent stored in contacr wj-th rire tgfj!-jgy neual of Mll- bomblcti aE 4 oC.
t+-2

tl

qqIiJ fl {- t..-,1 l.!-'i-.:. lClr

100
90

80
70

60
50

340
U)

F.i

r-{

u30
OJ

q{

o !

X

o

&20
6,' Fl

o

,-o
F{

(!

l0

I

9 8 7

6

Storage (deys)

Figure 4-1 (u). ]:-"uiliry

Decalr ruhen

srored ar 4 o ro 6 og.(1a)

tlN

ci--{isil'l[L)

i

at Fort DeErickfn 1-951. The"iSnc \{as stored in glass containers at :50-oC -The-aasurts Jre prtsented in Table 4-2' A regression analysis was mad.e of these data, however, the slope of the regression equation not \,ras not signif icanLly different fror'r zerol i'e'; the decay \..ras
significant over the time interval in question'

(4)

(-'i)

.l\

16 week storjl-g--e- sqgb-il!cv- studv on NX was conducted

I

I

I I

(5) (U) The parameEer values' for the exponential viability decay constant (^ relaged to st.orage tj-ne for ltse in Equation (4.1) are presented in table 4'3 fot four different storage temperagures. Ninety five perce:1t confid.ence 1i:nits based on production plant agent could oC. in glass containers. c be derived. only for agent lL{ stored. at 4 Co 6 The other K, values were obtained from sunnary information reported in the referenEed d.ocuments in which confidence limit values were noc provided.. It is not knovrn why poorer recoveries were obtained at oc' -tS oc. Lhan at 4 oc. or 20

I I I I I I
\

{l{ Cl.,r.)Siir I [i)

F

UHCLASSIf

IL.IJ

Results of the effect c. (U) Effect of Storage on Infectivitv' of storage time on infecEivity of agent NX as reported by Pine Bluff 61""n.1(r4) on production runs of agent NX are presented in the last, colunrr of Table 4-1. An analysis of these data showed thac the correlation coefficient was noL significantly different from zeto, and EhaE too few data i.rere available ro estimete infectivity as influenced by storage time.

d-

(c) Efiecc of Storage on Munition Efficiencv and Aerosol Decav Rate

(f) (U) Fort Detrick conducted a series of tests in 1959 to compare the efiecr of storage on the aerosol viabil,ity 31d i_r1f_es!iyi-!_y parameter of_-egerqt IrX s-tored in @sus-?g.g-tt! a!qf-"q "glaJ]-containers. NX from the same production lot was stored in these

fl

it
H {l !.

ri

4
ili fr

cffitervalsofl0and6Odays.Thestoragetemperature ras 4 oC. The agent stored in glass containers was disseminated by a PT-L2 tesE fixiure in a tesl chamber and the agent in the bomblet r,zas aerosolized b1- firing the 11114 in the test sphefe. The ternperature in bo;h the spie:: and the chamber was 24 oC. fZS or.) and the relative humidity was estimated at 85 Percent. ABP-30 samplers: were Placed ac four stations and one minute aerosol samples were taken at 4, 18r 32, and 46 minutes after aerosol release. Guinea pigs of the Hartley strain ranging between 240 and 375 grams body weighL were used to ascertain infectivity decay.

::
i

\

t *
t
*

I

/_a

t

hi cl. A5.\i I- I i.ll

---

afFr dissemination

as the

o-t. ,Dissemination a. (U) Source Streneth.
n

lsms ,v

Munition source

s

trens

th

is defined

cloud Tffii-ediately

Effi'n 5 raicrons. Source strength is a function of the concentrat ana--TEE-quanEity of agent contained in the aerosol d.isseminating device and the efficiency of che device. lrssemination effic ator fraction (usuallv expressed as perc

t

i.nffi

ort"

Aerosols of NX can be disseminated as a point source or as a line source. A point source may be instantaneously generated by a munition such as an explosive device, or m4y be continuously generated for a finite duration. The various models used to determine the source strength of a munition with agent NX are defined as follows:

(1) Point source. wet slurrv. The source strength for a point source of wet agent disseminated by an explosive or other mechanism (with very short time duraEion) is.
:l u
a:l ir iii

fl :1
i4

Qo =

VG.,rd

(4.3 )
by

where

QO

= source strength of the agent cloud formed
organ].sms

the munition,
m1,

Y

i \

s
F

t l
t
T

V = volume of agent slurry (fill
Gv

in

Ehe

munition),

= concentrat.ion of viable organisms in the munition fill at, tirne of unrnition funcEioning, organisms/m.g

T
F

0 = efficiency of the umnit,ion in convert,ing agent fill into a viable agent cloud of particles less than 5 microns in
diameter, dirnensionless
.

source

(2) Line source. wet slurry. of wet agent is
Qr=
EGv0
S

The source strength

for a line
(4.4)

where Ql = son.ce strength of the agent cloud per unit, of line source lengttrfi organisms/m E = rate of agent emission from the device,
4-8
m'0/min

1

'\t l.l,

,t_s SlFI

S = speed of delivery vehicle carrying the dissiminating

device, m/min

G.r0 =

as defined above.

(3) Line of point scurces. A line of relatively closely spaced. point sources (bomblets or generaEors) may be treated as a line source when the effecl-s t.o be considered are at sufficient distances dor,mwind for Ehe individual point source clouds to merge. In this case the source strength is Q/ = (number of point sources) x line length
Qp

(4.s)

The distance at which substant.ial overlap of the individual clouds generated by inst.anEaneous point source munitions will occur is given

in Figure 4-2 as a function of source spacing for various atnospheric stability-windspeed condiEions. Predictions in this graph are only true for level open terrain and fqr suull munitions. CorrecEion factors are available for larger insEanLaneous point source munitions or for source generators.(r e;

I

I I I I I

(2) (U) Munition efficiency data were not obtained in the Dugrvay Proving Ground field tests. However, estimat,es of Ehe muniEion disseminat,ion eff iciency wirh aeenrmbt F;aE ff;i"k ;; sonie-qf-EhE1e nfinitions .Gl:t J)- - rtr".E-.r ti*"t.] riln .ge"i r ir r vor,mu and source strengths are sho\.m in Table 4-5. The estimat,es are to be used only as a guide for an expected range of dissernination efficiencies and soLrrce strengths for agent NX. confidence limits were not provided vrith these estinates.

/,

-o

rraJ/-l r lqlL-if f'l t"r-. i-] -'r?
.-.

-t-'*' '-

I'IiCLASSIFIED

tion of disseminator efficiency. The cloud is sampled over periods of time beginning usually at 4 minutes. The Z time value is determined by fitting a least squares regression line t.o the 1og percent recovery of agentr or Particl-es, versus tirne of sampling, and extrapolating to time zeto. In a given test, the estin'r.ates for each of the replicate trials are pooled to obtain the geometric mean for dissemination efficiency. Confidence limits are calculated from Lhe standard error of estinaEe of pooled replicates. The antilogari.thm.of the mean logarithm percent cloud recovery at Z time is the geometric mean dissemination efficiency. (6) (U) The dissemination efficiency measurements made in the test chamber for these munitions provide insight into the range of dissemination efficiencies which can be expected from a variety of munitions. Dissemination efficiencies of the test fixEures involved in the NX chamber tests have been grouped according to the relative humidities at which they were tested. (Table 4-B). These data indicate an increased efficiency for NX disseminatioa at lower relative humidities.

(5) (U) In chamber trials, the compositisn of the aerosol cloud at time of dissemination, time Z, cannot be measured for a determina-

{l'FCl.r\S.iIpIff-i

b. I f*lu.imental Chamber Decay Data (f) (U) Most of the test chamber data available on lX has been generated from ForE Detrick tests conducted beEween 1954 and 1960. Althougir the weapons sysLem:i tested are now obsolete, a review of the Ceca',' data will yield insights into the IlX decay Par.ilr,eters. Table 5-i 1is ts the Fort Detrick tests r..-ith agent NX, the test condltions , and
5-2
t

JN

Cl . a 5-\l li- I t:0

unffi
the reported total decay constants. rn each case the agent employed was produced by Pine Bluff Arsenal or Fort Detrick and was freshly produced except for Tesrs 708 and 1127. storage times for agents in these tests were 60 days in Tesr ll27 and varied from 0.5 to 4.0 months in Test 708. Results of t e!-C4_$g!_!!re decay raLe in six tests conducLed with the -L2 h of 85 percent was 2.0 _nr lel.=niLuEe. 'lne rate ln tour
s

perc ent

per

m!

percent relative hurpidity with the pT-12 average total dec

I

I

(3) (U) Chamber tesLs were conducted in England, in 195g, ro assess the aerosol stability of B. suis using the radioactive tracer method to assess physical Cecay.(_"2) the test chamber uscd r,,as the Toroid; the disseminator, the Collison spray device. The test encompassed a wide range of temperatur'e 5id relative humidity combinations. The ratios of recoveries of viable ce1ls over radioactive tracer cells were fiEted to Equation (5.4) by least squares. Table 5-2 contains the resultant calculated decay parameter for each set of conditions. A siurilar table appears in Operations Research Group Report 2L.(2o) Minor differences occur between the decay values. (4) (U) A regression analysis has been performed rvith the data (70 oF.) presented in Tabre 5-2 employing the biological decay rate as a function of relative hwnidity. This ana:lysis is presented

I I

inFigure5.1andindicatesasignificant1yro*effi _r_._-----t---:-:-rela-tine liumidltfa;. - Sufficie"r d;T]: -arc-n-

I

I I I I I

orm such an analysis using temperature as the variable. A detrimental effect of higher temperatures on aerosolized B. suis survival is evident, however, in lhe temperature relative humidity matrix shovm in Table 5-3. This table contains half-1ife survival times and was constructed from values presented in Table 5-2.

-

5-5

1.0

..{
H

,x

u

t\

X
X t. oB)

;;c
IJ
L

O r{

c)

:>

up
\TX
I

u
(!

OJ

.t\

o

o

::t> ,'Jl'

+i

o
0)

i_rl
li/ r=

a

i,r--.

40

50

60
(7.)

RelaLive Humidity (RH),

Figure 5-1 (U).

w

Biological Decay RaE,e as a FuncLion of Relative Humidity aL 21 oC. (70 oF.) Measured ln Ehe Britlsh Toroid . ( ae;
NX

'i
CHAPTER 6

AGENT EFFECTIVENESS PREDICTIONS (U)

I

I I I

aerosol
dosage.

(1) (u) (2) (u)
dosage

Casualty rates as a function
Time

of B. suis aerosol

dosage.

to casualty (response) as a function of B.

suis

(3)

(u) Severity of response as a function of

suis aerosol

I

(1) (U) Lengtli wf incapacitation as a function of B. suis aerosol dosage. b. (U) Cfinical response to B. suis is highly variable under conditions of natural infection. This is probably due to the wide variation in the route of infection, in exposure dosages for individuals, and in the susceptibility of these individuals to B. suis organisms. 6-2. (U) Casualty Predictions
The general casualty prediction model for biological agents is presented in Volume X of the Source Book. When and if data sufficienE to accurately predict the effects of B. suis on a population become available, this modeL may be utilized for casualty prediction.

I I I I I

I I I I I

downwind dosages when the agent is disseminated at a flow rate of 20 gallons per minute from an aircraft traveling at the rate of 500 knoEs 6-1

(2) (U) Estimates presented in Figures 6-1 through 6-4 depict

w

i IH

Cl. A.5.\iir ll.L)

U tI 1-LArrrlrr

rtas

The dovmwind dosages for agent witlr. q d_99ay:5,qt_e of 1.5 P_etge_I-I-c P,e_r mTnutE-whdre --relEase-w-as-aE-a helght -o f.- 75 pr l-5O-m,e-t.eF-s- ll-e- p.l_otted

mee-ers-pei_je!-b,-ridr.

tir .^- wind speed-o-f-9 met-ers-iet Ce-.-g"d--1$a reSpectively. Figure 6-1 depicts dor'nwind dosage i"ii"g-Z neutral meteorological condition and Figure 6-2 depicts dosages unaei-5oth neutral and inversion conditions. Downwind dosage is not depicted for-inversion conditions at 9 meters Per second rvind speed since this condition normally does not exist. Figures 6-3 and 6-4 represent downwind dosages under the same conditions as Figures 6-1 and 6-2 except that the agent decay rate is 3 percent per minute. a u s e th i s c on d i t i on D o s a g e s a r g_ry!_ll€gig!=q i :_919 i5_i__ol !J.
A:2.
" "j_31::^ gend;I1yaoes_aq-t_c4!1t'=.F_-.jrlg!!:1v1re'-'jelea-se5-of-l=-i't],i9-y_gy]_a:ouo. to--be conAuc?;e to avoid the'aefeterious effect of sunlight.

i" rfg;r;;'3-T;;a-

6-6

.t
J
-t

rIN

U

NLt'R')rrrr re.*
CHAPTER
7

DEFENSE (U)

7

-3. (U) Prophylaxis and Therapy

human immunization

At the present time there is no reliable vaccine available for against brucellosis. Tetracycline, administered jointly with streptomycin, has been shor+n to be effective chemotheraPy against brucellosis. a. Immunization (l) There is probably some degree of natural immunity present in geographical areas where Brucella are present in animal reservoirs. However, naturally acquired reslstance to brucellosis is relafive, since reinfection is common, and innnunity is not assured by exposure to the agent or by the presence of circulating antibodies, as measured by agglutinin, precipitin, opsonin, or bactericidal tescs.(3? '3o)
meng

(2) Considerable research has been directed toward the developof a vaccine which will confer a Lasting inrnrunity against brucellosis infection. Inununity following natural infection or vaccination

with strains of aLtenuated virulence can be overcome by exceptionally virulenc strains or by massive exposures to the agent. Evidence of reinfecEion in humans has been neg"6.(z) (3) The Russians rePortedl-y have subcutaneously irmnunized over 3 million people with att,enuated virurent B. abortus, stralns BA-r9 andREV-1.Itisc1ainedthatthismassivelaffi''campaignhas resulted in major reduction in the incidence of brucellosis in humans.(se1 Although the BA-19 vaccines are reported to be highly effecEive, there are no published quantitative data concernlng the leve1 of protection afforded by these vaccines against bruceliosis infection, i.e., challenge studies with Brucella Ln inrnunized. individuals. (4) Spink, and othersr(40) conducted experirnents in humans using attenuated B. abortus strains BA-19 and REV-I. It was noted that 25 percent developed undesirable sequelae following the use of BA-19, and signs and symptoms of acute brucellosis appeared in 69 percent of the volunteers inoculated with the REV-I vaccine. Four men in the lacter group were ill enough to require hospitalization. rt rvas concluded that, at present, neither yaccine _(BA-19 or REV-I) appeared to be safe for utilizatLon in general lnrnunization programs. This was essentially the same conclusion that Pappagianis, and others,(41) stated as a result of their vaccination studies in 1965 with the REV-I s train. (5) Ce1l wa1l products and kilted whole cel1 vaccines prepared by ullrasonic inactivation have been tested for their inrnunizing effectiveness. A11 preparaEions gave encouraging results in the mouse and guinea pig, buE further study is needed before their value against naturally and/or artificially acquired infection in man is knovm.(4"'43) It is concluded that there is presently (1970) no safe and reliabLe method of immunization againsL brucellosis. b.
Chemotherapy

(1) The treaunent of the natural infection depends upon the seriousness of the disease and the presence or absence of a toxic state.(+z r+s ) No studies have been conducted with humans with regard to the efficacy of chemotherapeutic agents in reducing the casual-ty rates that might resuLt from an aerosol infection. It is probable that the chemotherapeutic agent employed to treat naturally infecEed individuals r^rould also be effective against an artificially produced disease. (2) General treatment for brucellosis consists of bed rest, nursing care, and supportive measures. Chemotherapeutic agents such as terracycline, streptomycin, chloramphenicol, novoblocin, aureomycin, terramycin and sulfadiazine have been used both singly and in various combinations to treaE individuals.(1142) A combination treaEment r,rith tetracycline and streptomycin has been found to be most

,r*ffi

7-2

TINCI-ASSITTED
effective.(1'41,42) Table 7-1.
Reconrnended

drug combinations are listed in

(3) The Joint Food and Agricultural Organization- World Healrh Organization (FAO/WHO) ExperE Conrnittee on Brucellosis has reconrnended tetracycline and sEreptomycin as the antibrucella regimen of choice. rn brucellosis of the bones and joints, it is suggested that 500 to 750 milligrams of tetracycline be given orally every 6 hours and 0.5 grams of streptomycin be administered inEramuscularly twice daily for 4 weeks. streptomycin at such d.osages only rarely causes damage in vestibular or auditory function of the eighth cranial nerve.(44) (4) In some cases of brucellosis, an endotoxin is released. into the body within L or 2 days following the initiation of antibiotic treatment.(43) End,otoxin liberation may cause vascular collapse and. secondary shock.(z) For the toxic state of illness, corticosteroid. therapy administered simultaneously vrith antimicrobial therapy is advised. Suggested oral doses are presented in Table 7-2.

Table 7-l (U).

Reconmrended Chemotherapy

suis Infec tioris

for Brucella

Dr

Combin

IONS

Minimwn Treacment LeneLh davs

Tetracycl ine
S

treptomyc in

2.0 (intramuscularly)

0.5 every 6 hours (oral)

y4(+z)

Su1
S

fadiaz ine treptomyc in

6.0 (oral) 1.0 to 2.9 (intramuscularly)

yo(++)

TabLe

7-2

(U)

Recormnended

Corticosteroid Therapy for the Treatment of Antibiotic Toxic Reaction
Dose

Daily
u

(me)

Frequency

of

Dose

Length of TreaEment (davs)

Cortisol
isone Predn is one
Cor t

100 300 20

Every 8 hours Daily Every 8 hours

or daily

3 ;s 4(+z) 1 ge 3(aa) 3 to 4(as)

7-3

trINCLASSIFIED

UNCI.ASSIFIED
7-4 (U) Decontamination
Beta propiolactone (BPL), ethylene oxide and other bactericidal chemicals may be used to kill Brucella suIs.
Decontaminat,ion of Brucella on skin surfaces can be accomplished by washing with cormnon disinfectantsr e.8., 2 percent lysol solution or 0.5 percent phenol solution. Pasteurizat,ion will kill che organisms in water, miLk, and various foods.(?)

a.

hours

B. -Ilt2 will vriths tand exposure to direct sunlight for 4\ suis ho,E exPosure to 2 percent lysol solution is required to kill the organism in soil.
b.
.
(

o6T

c. Decontamination of equipment, buildings, st.ructures, and of surrounding areas contaminated with any of the Brucella has not been investigated. However, since the organisms do not form spores, it can be assumed that BPL, a highly effective bactericidal ageng,(4zr4a) would be an effective decontaminant. The decontamination of large buildings rvith BPL will require recirculation of air throughout the structure. This will not be possible without a built-in recirculation sysCem, although in some structures.the,air can be recirculated by means of a system of fans. If the strucfure has many sma1l rooms and lacks a recirculating system, a number of smal-l BPL dispensers can be set up throughout and fans employed to assure adequate distribution.(as) Th" decontarnination properties of BPL have been extensively discussed in Part One, Volume VIII. d. Ethylene oxide, a bactericidal gaseous decontaminant, can be most effectively employed for decontaminating convoluted articles or hard to reach contaminated spaces. Smal1 objects (books, tools, shoes, uniforms) can be placed in plastic bags in which ethylene oxide is released.(so) There are no data to indicate the time and concentrations necessary to decontaminate articles contaminated with Brucella.

7-4

T'NCI-ASSIFIED

,ry
b. (U) Storaee. Additional data are needed on the effects of prolonged sEorage on the viability decay, infectivity decay and aerosol properEies of plant produced B. suis. These effects have not been inEensively studied on frozen production 1ot agent stored for prolonged periods. The possible deleterious effects of repeat.ed rrfreeze-thar^r" cycling on sEored agent viability and infectivity have not been precisely evaluated. Infectivity decay and aerosol properties are not oC. No reliable fully established for production lot agent stored at 4 information exists relating to the effects of storage on the aerosol properties of the other strains of Bruce11a.

8-2

i-r t:

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