k&,5.[.5 i I

ffir-,$_lflIInI

'i

I

eo-b' /4'? ryRDTE Project No. I -X-6-65704-D -634-02USATECOM Project No. 5-C0-473-000-016

DTC Project No. DTC 7l-504

*lnl/..

I rl.'rr "f\:

\lt.'{

MAY I 97I

b L'-/Drstrib'tion d^rrro to u. s . ffi^"^,agencies only. (Test and Evaluation).

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

L'OINT CB TECHNICAL DATAs ouRc E BooK tul

VOLUME VI I I

Bacterial Diseases (U)

Part Three: Brucellosis (U)

HEADQUARTER,S . DESERET TEST CENTER ' FORT DOUGLAS, UTAH . 84I I3

Vav l97l Other requests for thisdoculent must be referred to: Corunand-ing Ceneral , neseret Test Center, FortDouglas, Utah 811 13.

l

$f\i.\ oF 172 COPI:

DTC 7l-3:.C

COPY

tINCI..A,SSIFIED

FOREI^JORD

This documenE was prepared in compliance with Depar::E:-:letter, "JoinE Contact Point for Chemical-Biologica- lIData," 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 volurrsaddresses 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:=

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.

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

Commanding GeneralDeseret Tesc CenterATTN: STEPD-TT-JP-I(S)Fort Douglas, Utah 84113

ParameEer values with confidence leve1s derived frc::::-- --::,:::::='and chamber test data are presented. Models and su:=:,:=-= =:: :'-,:::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.

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-:lAll 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.

UNCI-ASSIFIED

SNou'\SS\sWF ierrre

4-l \X Vtablllty Decay

ILLUSTMTIOT*S

Title Paqe

when Stored at 4 o to 6 oC. 4-a

i'2 Tl,e Relatlonshlp Eetween Hunltlon Spaclng and Dcnrn-wtnd Dtstance to Cloud Overlap . . 4-10

5-t trX btologtcel Dgcay Rlte as a Functlon of RelaclveHurnldlty 8t 21 oC.' (ZO o9.) Measrrred ln the BrlctshTorold . . 5-7

6-l !- suls Dormwlnd Dosage Prediettons for AertalReleases Durlng Wlnd Speeds of 9 Meters per Secondw!.th a Decay Rare of 1.5 Percent per Mlnute. . 6-2

6-2 B. suls Donnwlnd Dosage Predlcclons for AerielReleases Durlng l{tnd Speeds of 3 Meters per Secondwlch e Decay rl,ate of 1.5 Percent per Mlnute. . 6-3

6-3 B. suts Dorrrrwlnd Dosage Predlctlons for AerlalReleasee Durlng tltnd Speeds of 9 Feters per Secondwith a Decay Rate of 3 Percent per Mlnute. . 6-4

6-4 B. suis Donnwlnd Dosege Predlctlons for AerlalReleaaes Durlng Wlnd Speeds of 3 Meters per Secondwith a Decay Rate of 3 Percent. per Mlnute. . 6-5

{;

Q,r ; -*r-u-l \esg"*E-'

- ,. l frf"s'qc33 t'

, Jt'

!

II

ri^i^a n --*-3'

flrlF*-TABLES UruO[ASSfFilECI

TltleSymptoms of Hunran Brucellosls. . .

Inctdence of Brucellosls tn Occupsttonal Groups for

Page

3-3

Table

3-l?-t

4-7

4-8

4-1

1-2

4-3

4-5

4-6

4-4

1949 ln Mlnnesota, tZ\ and l,tlsconsln . . . 3-5,/t

storage Stabttlry "f lx/.. 4 o ro 6 oc. 4-3

\-/ oStorage Stablllty of NX ar -50 -C. . . . . 4-5

Viabl1lty Decay Paramecer Values for Agent }IX 1nStorrge.....4-s

Effect of Storage for 10 anl 60 Days on t{X Dissentna-tl,o'n Paraneters. . . . . 4-l

Estfuoated Efflclency for Munltlone Teered wlrh Agenr NX 4-ll

Esllnated Paraneter Values for Standardlzed BlologlcalMunltlons for Use wlrh NX. 4-llDlsseolnatlon EfflctencLes for M( Chaurber Trlals atFortDetrtck. ...... 4-13

Suooary of Test Flxture Dlsseolnatlon Efflctenctes for}tX Chanber Trlals at Fort Detrtck. . . 4-L4

Sumary of Decay Data fro@ Fort Detrlck Chaober TesrsConducted wlth l0(. . 5-3

Blologlcal Decay of l{X as Measured in the BritlshToroid Test Chasber. . 5-6

5-3 B. suls Half-Ltfe as a Functlon of Dtfferent RelarlveHnoldlty and Tenperature Cooblnatlons. 5-8

5-4 Calculated Blological Decay Cqastants for Collabora-ttve Tests 4, 4A, and 48 . . . . . 5-9

6-1 Esclnated Dc'ynwlnd Dtstance of E\o Produclng Dosagesfor B. suLs Releesed froo an Aerlal Llne Source. . . . O-l

7-t Recmrended Chemotherapy for 3rucelle sutc Infectlons. 7-3

7-2 Reconoended Corticosterotd Therapy for the Treatnent' of Antlblotlc Toxlc Reactlon . . l-3

A-l Fteld Trlals Conducted at DugLray Proving Ground A-lxL

5-l

5-2

UT{CLASSIFIET)fr

(u)

CHAPTER 1

I r**"t-t.OGeneral

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.)

d. (U) Duration of Illness. Variable; averages 3 months'(Inciudes all rhe Brucella.)

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 untreatedpatients vrith B. suis infecLions.

a. (U) Prcduction Slurry Count. 3:'.Ldo organisns per milli-'1

" r^-! ! LE! r

ilr)F

!\

I

I

I

I

l

I 4

pa t ient s

'f r-! r

l-1

I

UHCLASSIFIED

Storase Half-Life. 150 to 169 days at 4 o ro 6 oC.

Dissemination Eff iciency.

A/B45Y-1 and Aero 14B aerial dissemination tanks.Estimated Eo be 3 percent.

(2) M143 bomblet. Estimated co be 0.7

b. (u)

c. (u)

(l)

T

percent.

t-5. (U) Infectivity Decay

Unknor.m.

t-U. t

Effectiveness Predictions

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

(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 strep-tomyc::t is effective in reducing severity and duration of illness.

e. Decontamination. Beta propiolactone, etirelyene oxideand o:rer bactericidal chemicals are effective.

I'tl{ {-

L-2

IIIIIIIIII

uiIt-LA))rlu

-

CHAPTER 3

I o.u*t AND DTsEASE cHARAcrERrsrrcs (u)

3-1. l) General Properties

The three species of Brucella are characterized in Bergeyrs ysnsal(2)on the basis of antigenic properLies, tolerance to a variety of dyesrvhen gror!,n on sulici media, and various ocher determinative techniques.B. melitensis was the first organiqm identified as the causaLive agentof brucellosis and was shoqrn t,o be transmitEed from infected goats toman through ingestion of milk. Subsequently, B. abortus rpas isolatedfrom cattle, and B. suis from swine. These species have been found inmany dcrnestic and semid.oinestic animals and in man. Some strains have

been found. that are not readily assigned to any of the three designatedspecies. Among the many strains obtained fro-r:r various animals and

collecEed. in d,iverse geographic locations' a range of cultural andpachogenic characteristics have been demonstraEed within the confinesof the d.eterminaLive criteria of the genus.(e)

The Brucella are gram negaEive, nonsporulaling and nohmoCile. Theyare aerobic. However, soine sErains, particularly of B. alggg5r requirean aEnosphere containing approximately 10 Percent carbon dioxide inorder to grow in culture media after initial isolation from animalEissue. 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-

^---rL -^..] "9_. _a19 rec9gnr14!_lg_c=*tg may beure is 37 "C. and recognizabl

3-3. D Strain Selection and Developrnent(a)

3-2. (U) Biological Nature of the Organism

I

3-1

fr)i<f".

TJilCLASSIFIED

..

i.

3-4. (U) CharacEeristics of the Disease

a, Nornenclature. The disease produced in man by theBrucella is brucellosis. Synonyms used to designate the dj-sease areIlalta 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, infec-tious aborE.ion, and epizootic abortion.

b . Transr,ris s ion. Bruce I Los is is character ized b1z extens iveinvasions o' tissues of Ehe body by the infective organism. The organ-isms remain in the tissues for long periods. In animals, where theinfection becomes latent, it may persisE for years. The Brucella arepassed from rnfected animals, in milk, and in feces, urinE-ZnE-6Eher bodvexcreta. The disease in man is most frequently found as an occupa-tionally related disease of farmers, neat handlers, veterinarians,and others ergaged in handling aninals or in processing animal pro-ducts.\! ':'

c . S!-::.p t.oms

(i) lhe onset of brucellosi-s uray be abrupt, manifested by chills,fever, and sweati-ng, ,or it may be an insidious development. manifesledb.7 weai<ness and mild incapaciEation over an extended period. Table3-l- fists sl.rnptorns chai are predorninant and the frequency with whicht:lel/ may cccur . The nani f es taEions and assoc ia teii severity of s1'rnptomsi,-,-e been -:sed to characte::ize f lve distieeuishacie types of thel:-sease i:. l'tumans :(: )

(.3) InterrlittenL :\'De. Assoc:-aced witi siriiting articular

III

I

j

J-J

UNUIJ\SbIf IET/

Table 3-1 (U). Symptoms of Human Brucellosira(z)

Number Percent of otal

FeverChil1sMalaiseWeaknessBody acheSweatingHeadacheAnorexiaWeight loss

B 9.573.37L.767 .O67 .067 .O51.839.336.6

aProvisional data from 191 case reports in 1968.

L7L140L37L28128128

997570

\

\

rireumaEisi,r, !.'eakncss, ncc:ur::al p.3:spiration, anC a_ temperature nearnormal in the morning, but rising to 101 " to 104 eF. in the evening.

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

(c) Undulant type. Characterized by daily fluctuations in thebody temperature generally caused by B. melitensis.

(d) Malisnant tvpe. Sustained high temperature becomingexEreme prior to death. Almost always fatal, generally caused by B.melitensis infection.

(e) Atvpical chronic type. Manifested clinically by muscularsEiffness, gastric disturbances, and various neurological abnormalities.

(2) BronchiEig may be associated with infection of the organisnsthrough inhalation. ?ulmonary invasio+ may also be associated withinfection t.hrough other modes of entry wherein the lungs are invadedas a consequence of proliferation of bacteria within the body. Thenature of Ehe sJnnptoms for any given case of the disease canno! bealigned 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 thedisease.lt r= re)

d. Onset and Duration

(i) Variation in onset, or incubaEion time, and in length of

ITNCI-ASSIFIEDi

,,.-.,r. =.sl*iGtn{raEH

UNCI.^H,SSIFIED

illness has no apparent relafionship to moi.e of infeccion or to Chestrain of Brucella involved, as indicated by the variability observedin the outbreaks of the disease. The incubation period of the diseaseranges from a few days to several monEhs, with andays. In the case of laboratory accidents with B.sis, the majority of cases have had an incubat.ionB weeks.(z)

average of 8-10suis and B. meliten-

period between 4 and

(2) Studies of brucellosis case histories have indicated a dura-tion of debilitation serious enough to require hospitalization rangingfrom 20 days to over a year. LaboraEory acquired infections showed thes€rme general variability in duration to those by 'hatural'r means. Whenthe disease is established as a chronic infection, Ehe debilitation mayrecur intermittently over a period of years.(s) The average incapaci-t.ation period has been estimated to be 3 months.(e)

Susceptibilitv and Severitv

l

'

(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 tobe extensively exposed to the organisms. In one grouP of 120 veEer-inarians, where Lhe probability of frequent exposure to infectiousmaterial exceeded 90 percent, only three indicated any clinical hisEoryof brucellosis, although 45 percent had agglutinins for the Brucella'One of the group was exposed Ehrough ingestion and skin abrasion, butdid not dev-lop clinical symptoms of the disease or agglut,inins forthe Brucella. In another Sroup of 49 vet.erinarians, 75 percenE had

.ggfffiitr" for Ehe Brucella and Ehree had a history indicatingclinical symptoms.(s'6)

(2) Incidence of brucellosis in occupational groups in fourMidwestern States is shoqrn in Table 3-2. No precise assunption as toexposure rate for these groups can be rnade. However, it is believedthat the rate was high.(s)

(3) In sunrnarizing these data, it was concluded that in a popu-lation where the probability of exposure to Brucella is high, theyearly incidence of clinical brucellosis will not exceed 5 percent ofthe population, and would more likely be approximately 1 percent.(5)Other case studies citing specific Brucella species have indicatedmuch higher susceptibility. (") Susceptibility of B. melitensls has beenreported as high as 40 percent.

(4) In an accidental exposure of students to what was assumed tobe an aerosol of B. melieensis, it was reconstructed that a certaingroup of individuals was exposed to a high dosage, and anolher to alower dosage. In the high dosage group, 25 percent were clinically ill,and an additional 30 percent indicatec no symptoms, but did dcvelop a

?-.

g'I

1

I'I,Iti

I

T

IINCIS.SSIFIED

UilCLASSIFIED

pos itsive irrrnunological response.percenE were clinically i1l andposiE,ive irrrnunotogical response.

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

Table 3-2 (U). Incidence of BrucellosisL949 Ln Minnesota, Iowa,

in Occupational Groups forand. wisconsin(6 )

\

S Eate

VeLerinariansPacking House

WorkersFarmers and Ani:nalHusbandrv Workers

Total I TotalPooulation I er.o=

TotalPopulation

TotalCases

TotalPopulatior

Total

IowaMinnesotaWiscons in

TOT-^.L

Rate (%)

740403502

L,645

9

42

15

0.9r

22,9L0L5,2358.497

a(),642

4L5026

il7

o.25

265,500249,700267 ,300

782 ,500

L73L4Ll1s

429

0.05

(5) Severity of illness varies markedly. Many cases are declaredsubclinical with the patient only slightly debilitated. The duration ofincapacitation may be long, but in many cases illness would not be soint,ense as to preclude performance of necessary tasks. To some extent,severity of illness is related to the species of Brucella, with great-es! severity resulting fron infection with E. melitensis. Severityand duration of the disease can be markedly reduced by treatment withantibiotics.(s '?)

(6) Variation in susceptibility and, severity of the disease withaB€r sex, and other population categories, has been found to occur tosome degree. The young are more resisEant to brucellosis than areolder people. The disease has seldom been reported in children under12 years of age except in undernourished populations. In such popula-tions, chfldren who contracE the disease respond as severly as doadults.( z )

f. Mortalitv and Seguelae

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

\tI

1iry c!, n 5.!rFIi:J

)

)t\

UT'ICLASSIFIED

(2) serious cornplications which m"y be persist,ent occasion-all-y acconrpany brucellosis. Recurrent chills and, fever, localizaEionor organisms in calcified necrotic lesions, episodic myalgia, granulom-atous reactions appearing as rElsses in the spleen and liver aresc'me of the permanenE sequelae. Central nervous system effects, suchas depression, frustration, anxiety, irritability, restlessness, an6apathy, are often noted. organic lesions of the cerebral vessels,meningo-encephalitis or meningitls may occur. other organs which maybe affected are the heart, (bacterial endocarditis), deep veins (granllom-aEous changes in vessel walls causing thrombosis), lungs (pulmonaryerobol-ism), reproductive systern (epididyuro-orchitis occurrLng in 2 to5 percent of adult males), kidney, ureter and bladder (granulomatouslesions, sonetirnes calcification), and the skin (rashes). The con-junctiva, cornea, iris, choroid, retina,.opt,ic nerve, and. ext,ernalmuscles of the eye also may be involved,.(6r7)

(S) InfecEivity

infectivity of B. suis has been studied infor 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

and the "rbffi"r"o.r ED"o for **k. -es_g_-lha! !pg_o:gar1!ggrs.(Ir)

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

3-6

ti

t-

If;

\;I

*^^.,,i:.I9l"if|iGTnestirnated to be 60

Fp c!-*^ 55lf lli)

fr

UHCLASSIFIE.D

CHAPTER 4

SOURCE PARAIETERS AND MODELS (u)

I

I

I

I

I

I

I

I

I

I

I

I

Agent Production

(1) (U) Holding temperatures.

(2) (U) Ntunber of freeze-thaw cycles.

conlainer material (especially the surfaces and coatings inrvitr the agenL slurry) may also influence viability.

I Effect of StoraEe on Viabilitv

(l) (U) For agent NX the reduction of viable cell concentrationat constant ternperature can be described by the exponen:ia1

deca,'r nodel :

S Loragecontac t

LV.

c t-nroJ

a'-o-2oae ev4 *Ce

I

!

Iil).,\..!)iFi

c(t) = co exp (-r"t)

the viable ce11 concentraEiona: time, t,

in cells per milliliEer

t::e initial viable cell concentration in cells perrn:lliliter at the time of production (t = o)

(4.1)

where c(t) =

-o

(^= exponential viabiliLy decays i.orage tirne , pe r da y

t = length of storage time, per

constant related to

The point in tine at which the viable cel1stored material- has {ecreased to one-halfcentration, Gc, is the storage half-tife,aad using equation (4.1)

concentration, G(t), in thethe initial viable agent con-Tso. By settinC c(t) - Go/2

Tso= o'd93K

S

(4.2)

I

t

I

:a-

I

Ii

(3) (u) rue data in Table 4-l were plotred as 1og percenr sur-vival versus storage age, and were fitted with a least squares regres-sion 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 dataI presented in Table 4-1 gave a mean decay oi tn

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 rhepreviously re2orred half-life of t50 days.(13) This longer half-lifenay be due i:: part to the storage of the agent in glass containers.tgfj!-jgy -aLf-1:-fe r-r4s re,ported.for agent stored in contacr wj-th rireneual of Mll- bomblcti aE 4 oC.

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

t+-2

100

90

80

70

60

50

I

340U)

F.ir-{

OJu30q{oX!

oo&206,'

Fl

,-o(!F{

l09

8

7

6

Figure 4-1 (u). ]:-"uiliry Decalr ruhen srored ar 4 o ro 6 og.(1a)

Storage (deys)

tlN ci--{isil'l[L)

i

(4) (-'i) .l\ 16 week storjl-g--e- sqgb-il!cv- studvat Fort DeErickfn 1-951. The"iSnc \{as stored in:50-oC -The-aasurts Jre prtsented in Table 4-2'was mad.e of these data, however, the slope of the\,ras not signif icanLly different fror'r zerol i'e';significant over the time interval in question'

on NX was conductedglass containers atA regression analysisregression equation

the decay \..ras not

I

I

IIIIIIII

(5) (U) The parameEer values' for the exponential viability decayconstant (^ relaged to st.orage tj-ne for ltse in Equation (4.1) arepresented in table 4'3 fot four different storage temperagures. Ninetyfive perce:1t confid.ence 1i:nits based on production plant agent couldbe derived. only for agent lL{ stored. at 4 c Co 6 oC. in glass containers.The other K, values were obtained from sunnary information reported inthe referenEed d.ocuments in which confidence limit values were noc

provided.. It is not knovrn why poorer recoveries were obtained at-tS oc. Lhan at 4 oc. or 20 oc'

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

i\

F

riitfl4H{l!.

ilifr

::

t*t*I

UHCLASSIf IL.IJ

c. (U) Effect of Storage on Infectivitv' Results of the effectof storage time on infecEivity of agent NX as reported by Pine Bluff61""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 corre-lation coefficient was noL significantly different from zeto, and EhaE

too few data i.rere available ro estimete infectivity as influenced bystorage time.

d- (c) Efiecc of Storage on Munition Efficiencv and AerosolDecav Rate

(f) (U) Fort Detrick conducted a series of tests in 1959 tocompare the efiecr of storage on the aerosol viabil,ity 31d i_r1f_es!iyi-!_yparameter of_-egerqt IrX s-tored in @sus-?g.g-tt! a!qf-"q "-glaJ]-containers. NX from the same production lot was stored in thesecffitervalsofl0and6Odays.Thestoragetemperatureras 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 inbo;h the spie:: and the chamber was 24 oC. fZS or.) and the relativehumidity 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 Hartleystrain ranging between 240 and 375 grams body weighL were used toascertain infectivity decay.

/_a

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

--

---

i\

a. (U) Source Streneth.as the nafFr dissemination

o-t. ,DisseminationMunition source

lsms ,v

Effi'n 5 raicrons. Source strength is a function of the concentratana--TEE-quanEity of agent contained in the aerosol d.isseminating deviceand the efficiency of che device. lrssemination efficfraction (usuallv expressed as perc atort ort"i.nffi

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

(1) Point source. wet slurrv. The source strength for a pointsource of wet agent disseminated by an explosive or other mechanism(with very short time duraEion) is.

Qo = VG.,rd (4.3 )

the munition,where QO = source strength of the agent cloud formed byorgan].sms

V = volume of agent slurry (fill in Ehe munition), m1,

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

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

(2) Line source. wet slurry. The source strength for a linesource of wet agent is

fl:1:lua:li4

iriiiY

stlF

tT

TF

S

where Ql = son.ce strength of the agent- lengttrfi organisms/m

(4.4)

cloud per unit, of line source

the device, m'0/min

Qr= EGv0

E = rate of agent emission from

4-8

SlFI

s trens t h is definedcloud Tffii-ediately

1 '\t l.l, ,t_s

I IIIII

S = speed of delivery vehicle carrying the dissiminatingdevice, 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 sourcewhen the effecl-s t.o be considered are at sufficient distances dor,mwindfor Ehe individual point source clouds to merge. In this case thesource strength is

Q/ = (number of point sources) x Qp (4.s)line length

The distance at which substant.ial overlap of the individual cloudsgenerated by inst.anEaneous point source munitions will occur is givenin Figure 4-2 as a function of source spacing for various atnosphericstability-windspeed condiEions. Predictions in this graph are onlytrue for level open terrain and fqr suull munitions. CorrecEionfactors are available for larger insEanLaneous point source munitionsor for source generators.(r e;

(2) (U) Munition efficiency data were not obtained in the DugrvayProving Ground field tests. However, estimat,es of Ehe muniEion dissem-inat,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,muand source strengths are sho\.m in Table 4-5. The estimat,es are to beused only as a guide for an expected range of dissernination efficien-cies and soLrrce strengths for agent NX. confidence limits were notprovided vrith these estinates.

/, -o

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

I'IiCLASSIFIED

tion of disseminator efficiency. The cloud is sampled over periods oftime beginning usually at 4 minutes. The Z time value is determined byfitting a least squares regression line t.o the 1og percent recovery ofagentr or Particl-es, versus tirne of sampling, and extrapolating to timezeto. In a given test, the estin'r.ates for each of the replicate trialsare pooled to obtain the geometric mean for dissemination efficiency.Confidence limits are calculated from Lhe standard error of estinaEe ofpooled replicates. The antilogari.thm.of the mean logarithm percentcloud recovery at Z time is the geometric mean dissemination efficiency.

(6) (U) The dissemination efficiency measurements made in thetest chamber for these munitions provide insight into the range ofdissemination efficiencies which can be expected from a variety ofmunitions. Dissemination efficiencies of the test fixEures involvedin the NX chamber tests have been grouped according to the relativehumidities at which they were tested. (Table 4-B). These data indicatean increased efficiency for NX disseminatioa at lower relative humidities.

(5) (U) In chamber trials, the compositisn of the aerosol cloudat 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 beengenerated from ForE Detrick tests conducted beEween 1954 and 1960.Althougir the weapons sysLem:i tested are now obsolete, a review of theCeca',' data will yield insights into the IlX decay Par.ilr,eters. Table 5-i1is ts the Fort Detrick tests r..-ith agent NX, the test condltions , and

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

5-2

I

I

I

IIIIIII

unffithe reported total decay constants. rn each case the agent employedwas produced by Pine Bluff Arsenal or Fort Detrick and was freshlyproduced except for Tesrs 708 and 1127. storage times for agents inthese tests were 60 days in Tesr ll27 and varied from 0.5 to 4.0months in Test 708. Results of t e!-C4_$g!_!!re decayraLe in six tests conducLed with the -L2

of 85 percent was 2.0 _nr lel.=niLuEe. 'lne rate ln tourpercent relative hurpidity with the pT-12 average

per m! total dec

(3) (U) Chamber tesLs were conducted in England, in 195g, roassess the aerosol stability of B. suis using the radioactive tracermethod to assess physical Cecay.(_"2) the test chamber uscd r,,as theToroid; the disseminator, the Collison spray device. The test encom-passed a wide range of temperatur'e 5id relative humidity combinations.The ratios of recoveries of viable ce1ls over radioactive tracer cellswere fiEted to Equation (5.4) by least squares. Table 5-2 containsthe 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 thedata (70 oF.) presented in Tabre 5-2 employing the biological decayrate as a function of relative hwnidity. This ana:lysis is presentedinFigure5.1andindicatesasignificant1yro*effi

-

_r_._-----t---:-:--rela-tine liumidltfa;. - Sufficie"r d;T]:-arc-n- orm such an analysisusing temperature as the variable. A detrimental effect of highertemperatures on aerosolized B. suis survival is evident, however, inlhe temperature relative humidity matrix shovm in Table 5-3. Thistable contains half-1ife survival times and was constructed from valuespresented in Table 5-2.

hs

perc ent

5-5

1.0

..{H

,x

uc)Or{

OJuupI\TX

(!o0)a

;;cIJL

:>::t>,'Jl'i_rli,r--.li/r=

605040

aL 21 oC. (70 oF.)NX Biological Decay RaE,e

Measured ln Ehe Britlsh

RelaLive Humidity (RH), (7.)

as a FuncLion of Relative HumidityToroid . ( ae;

Figure 5-1 (U).

w

X

X

+io

o

t. oB)

t\

.t\

'i

IIIIIIIIIIIIIII

CHAPTER 6

AGENT EFFECTIVENESS PREDICTIONS (U)

Casualty rates as a function of B. suis aerosol dosage.

Time to casualty (response) as a function of B. suis

(1) (u)

(2) (u)aerosol dosage

(3) (u)dosage.

Severity of response as a function of suis aerosol

aerosol dosage.

b. (U) Cfinical response to B. suis is highly variable under con-ditions of natural infection. This is probably due to the wide varia-tion 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 ispresented in Volume X of the Source Book. When and if data sufficienEto accurately predict the effects of B. suis on a population becomeavailable, this modeL may be utilized for casualty prediction.

(2) (U) Estimates presented in Figures 6-1 through 6-4 depictdownwind dosages when the agent is disseminated at a flow rate of 20gallons per minute from an aircraft traveling at the rate of 500 knoEs

6-1

w

(1) (U) Lengtli wf incapacitation as a function of B. suis

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_ottedi" rfg;r;;'3-T;;a- A:2. tir .^- wind speed-o-f-9 met-ers-iet Ce-.-g"d--1$amee-ers-pei_je!-b,-ridr. reSpectively. Figure 6-1 depicts dor'nwind dosagei"ii"g-Z neutral meteorological condition and Figure 6-2 depictsdosages unaei-5oth neutral and inversion conditions. Downwind dosageis not depicted for-inversion conditions at 9 meters Per second rvindspeed since this condition normally does not exist. Figures 6-3 and

6-4 represent downwind dosages under the same conditions as Figures6-1 and 6-2 except that the agent decay rate is 3 percent per minute.D o s a g e s

- a r g_ry!_ll€gig!=q i "j_31::^ :_919 i5_i__ol !J. "

a u s e th i s c on d i t i ongend;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.

.t-tJ

rIN

6-6

U NLt'R')rrrr re.*

CHAPTER 7

DEFENSE (U)

7 -3. (U) Prophylaxis and Therapy

At the present time there is no reliable vaccine available forhuman immunization against brucellosis. Tetracycline, administeredjointly with streptomycin, has been shor+n to be effective chemotheraPyagainst brucellosis.

a. Immunization

(l) There is probably some degree of natural immunity presentin 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 exposureto the agent or by the presence of circulating antibodies, as measuredby agglutinin, precipitin, opsonin, or bactericidal tescs.(3? '3o)

(2) Considerable research has been directed toward the develop-meng of a vaccine which will confer a Lasting inrnrunity against brucel-losis infection. Inununity following natural infection or vaccination

with strains of aLtenuated virulence can be overcome by exceptionallyvirulenc strains or by massive exposures to the agent. Evidence ofreinfecEion in humans has been neg"6.(z)

(3) The Russians rePortedl-y have subcutaneously irmnunized over3 million people with att,enuated virurent B. abortus, stralns BA-r9andREV-1.Itisc1ainedthatthismassivelaffi''campaignhasresulted in major reduction in the incidence of brucellosis inhumans.(se1 Although the BA-19 vaccines are reported to be highlyeffecEive, there are no published quantitative data concernlng theleve1 of protection afforded by these vaccines against bruceliosisinfection, i.e., challenge studies with Brucella Ln inrnunized.individuals.

(4) Spink, and othersr(40) conducted experirnents in humansusing attenuated B. abortus strains BA-19 and REV-I. It was noted that25 percent developed undesirable sequelae following the use of BA-19,and signs and symptoms of acute brucellosis appeared in 69 percent ofthe volunteers inoculated with the REV-I vaccine. Four men in thelacter group were ill enough to require hospitalization. rt rvas con-cluded that, at present, neither yaccine _(BA-19 or REV-I) appeared tobe safe for utilizatLon in general lnrnunization programs. This wasessentially the same conclusion that Pappagianis, and others,(41)stated as a result of their vaccination studies in 1965 with the REV-Is train.

(5) Ce1l wa1l products and kilted whole cel1 vaccines preparedby ullrasonic inactivation have been tested for their inrnunizingeffectiveness. A11 preparaEions gave encouraging results in the mouseand guinea pig, buE further study is needed before their value againstnaturally and/or artificially acquired infection in man is knovm.(4"'43)It is concluded that there is presently (1970) no safe and reliabLemethod of immunization againsL brucellosis.

b. Chemotherapy

(1) The treaunent of the natural infection depends upon theseriousness of the disease and the presence or absence of a toxicstate.(+z r+s ) No studies have been conducted with humans with regardto the efficacy of chemotherapeutic agents in reducing the casual-tyrates that might resuLt from an aerosol infection. It is probablethat the chemotherapeutic agent employed to treat naturally infecEedindividuals r^rould also be effective against an artificially produceddisease.

(2) General treatment for brucellosis consists of bed rest,nursing care, and supportive measures. Chemotherapeutic agents suchas terracycline, streptomycin, chloramphenicol, novoblocin, aureo-mycin, terramycin and sulfadiazine have been used both singly and invarious combinations to treaE individuals.(1142) A combination treaE-ment r,rith tetracycline and streptomycin has been found to be most

7-2

,r*ffi

TINCI-ASSITTED

effective.(1'41,42) Reconrnended drug combinations are listed inTable 7-1.

(3) The Joint Food and Agricultural Organization- World HealrhOrganization (FAO/WHO) ExperE Conrnittee on Brucellosis has reconrnendedtetracycline and sEreptomycin as the antibrucella regimen of choice.rn brucellosis of the bones and joints, it is suggested that 500 to750 milligrams of tetracycline be given orally every 6 hours and 0.5grams of streptomycin be administered inEramuscularly twice daily for4 weeks. streptomycin at such d.osages only rarely causes damage investibular or auditory function of the eighth cranial nerve.(44)

(4) In some cases of brucellosis, an endotoxin is released. intothe body within L or 2 days following the initiation of antibiotictreatment.(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 isadvised. Suggested oral doses are presented in Table 7-2.

Table 7-l (U). Reconmrended Chemotherapy for Brucellasuis Infec tioris

Dr Combin IONS

Tetracycl ineS treptomyc in

Su1 fadiaz ineS treptomyc in

TabLe 7-2 (U)

Minimwn TreacmentLeneLh davs

y4(+z)

yo(++)

Recormnended Corticosteroid Therapy for theTreatment of Antibiotic Toxic Reaction

0.5 every 6 hours (oral)2.0 (intramuscularly)

6.0 (oral)1.0 to 2.9 (intramus-

cularly)

uDaily Dose

(me)Frequencyof Dose

Length of TreaE-ment (davs)

CortisolCor t isonePredn is one

100300

20

Every 8 hoursDailyEvery 8 hours

or daily

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

7-3

trINCLASSIFIED

UNCI.ASSIFIED

(BPL), ethyleneto kill Brucella

7-4 (U) Decontamination

Beta propiolactonechemicals may be used

oxide and other bactericidalsuIs.

exposure to direct sunlight for 4\to 2 percent lysol solution is required

a. Decontaminat,ion of Brucella on skin surfaces can beaccomplished by washing with cormnon disinfectantsr e.8., 2 percentlysol solution or 0.5 percent phenol solution. Pasteurizat,ion willkill che organisms in water, miLk, and various foods.(?)

b.hours . (

to kill

vriths tandexPosure

in soil.

B. suis willo6T -Ilt2 ho,Ethe organism

c. Decontamination of equipment, buildings, st.ructures, and ofsurrounding areas contaminated with any of the Brucella has not beeninvestigated. However, since the organisms do not form spores, itcan be assumed that BPL, a highly effective bactericidal ageng,(4zr4a)would be an effective decontaminant. The decontamination of largebuildings rvith BPL will require recirculation of air throughout thestructure. This will not be possible without a built-in recirculationsysCem, although in some structures.the,air can be recirculated by meansof a system of fans. If the strucfure has many sma1l rooms and lacksa recirculating system, a number of smal-l BPL dispensers can be set upthroughout and fans employed to assure adequate distribution.(as) Th"decontarnination properties of BPL have been extensively discussed inPart One, Volume VIII.

d. Ethylene oxide, a bactericidal gaseous decontaminant, canbe most effectively employed for decontaminating convoluted articlesor hard to reach contaminated spaces. Smal1 objects (books, tools,shoes, uniforms) can be placed in plastic bags in which ethylene oxideis released.(so) There are no data to indicate the time and concentra-tions necessary to decontaminate articles contaminated with Brucella.

7-4

T'NCI-ASSIFIED

,ry

b. (U) Storaee. Additional data are needed on the effects ofprolonged sEorage on the viability decay, infectivity decay and aerosolproperEies of plant produced B. suis. These effects have not beeninEensively studied on frozen production 1ot agent stored for prolongedperiods. The possible deleterious effects of repeat.ed rrfreeze-thar^r"

cycling on sEored agent viability and infectivity have not been pre-cisely evaluated. Infectivity decay and aerosol properties are notfully established for production lot agent stored at 4 oC. No reliableinformation exists relating to the effects of storage on the aerosolproperties of the other strains of Bruce11a.

8-2

i-r t: