The hores and baa were exoined daily for the dcvelogment of clinical diseae

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Gift Vouchers - Gifts The hores and baa were exoined daily for the dcvelogment of clinical diseae

Proceclore Eightgrey-headed f.uit bals (Pie.opvspoliocephaius) werc inoculated and housed in contaci with thrce uninfecledbats and two rhinfected horces.In a second expeF iment,foilr horsesworeinocllatedby subcutan€ous injection and inllanasaiinoculaionand holsed in contactwith three unintected horsesand six uniniecled cats.In 6 find experiment. 12 cals were rnoculaled and housedin contactwith thrce uninlbct€dhorses.Two surviving horseswere inoculated at the conclusion of lh€ thid expeiment:the nrsi orallyand the secondby nasalsabbing.

All animalswere necrcpsied and examined by gross and microscopic palhological melnods, immrnoperox’daselo doteci viral antigen in iormalin-fixedtissues, vifls isolation was anempted on llssues and SNT and ELISA melhodswere us€d to detect Hev-sp€cificantibody.

Results Clinicaldisease was notobserved in lhefruitbals.

Althoughsix of eighl inoculatedbats deveiopedanlibody against HeV and 1woof six de\eloped !?scular lesions which contained viralantigen.

The in-conlact balsand hoEes did not serccon€ri Three of four hoses tiat w€rc inoculateddevel oped aalle disease.

But in-@nlact horses and cats were nol infected.In the thkd experimont,one of three in-contact horses contracteddisease.

At the tjme of necrcpsy,nigh litfes of HeV were detecledin lhe kidneysof six acuielyinfecred ho6es, in lhe urineof four horsesand the mouthol !eo, bui not in the nasal cavilies or t€cheas.

Conclusions Grcv-headediruit bats de\€lop subclinicaldiseas€when imculated with HeV Horses can be infeciedby o|onasalrcutesand can exclereHeV in urine and saliva-ll is possible to transnit HeV fiom cals to, Transmissionfrom P rolioceDralls lo horses could not be proven and n€ither could transmissionlrom horses io horses of horses to cals.

Underthe experimenl€lcondilions ot lhestudylhe virusis not highly contagious. ,4rsr ver J 1993:76:Sr 3.S13 orblrlivirus.

Nxi bri.

Hr* €r, wsure.

QeF edbe, l, ) o 4 .i n H e n d m” ,u b u -b o fB -1 b 4 r. ,.rc.Tn , : on wir h r p, e v ,o L .l vu n d e n ri b e dm e n h e – o i rh. | l-raanyrau nda.

Frnily cru\ed In ,hL derrh\ of I ho,.e, ‘ rn d I hum r nt r c mr r a u e ,e s p i d ,o .y d e $ r rT h e ” .u,.a.

Prov’sionally designredcquinemo.billivirusbur subsequent studisr’a indioted tlat rhevnuscouldnorbe*ilyclassificd in rn1 of tfc existing gcnela i.

Rhe fanily Pa/anyo,ifidae.

Consequendy the virushasbee! renamed Hendlavirus.rr I! ocrober I 99i a fa@q ddeloped fatal encephalitis s d rsult df HeV infection,6 whichq att’ibuledro exposure ro Mo HeV horseswhich had died morc than I ],earearlier.T ‘nfecred ,46r Ysrl voi 75, No 12, D€ember l9t8 lrrcnsive relolosical sur€ys ihoughou.

Qufenslandfound no evidence ofH.V infecrionin ho6esor hlmans-&tHowever lilh bar (flying tox6, spp)wcre fourd ro h.!t a hish “r@?6 preqleoce ofanribo4’ro Hev indiuring rhg’ tuayb€a wil.llife rcseryoi.

Ofthe virus.rd *idoce of HeV infeoion ha, noi beenfoundin mr.ninal species oAer thanrhese bas.

In e{perimerGl studics, guinea pigsand catshavebeenfound !o bcsus.eplible rc FIeVinf€ction’r’D The aimsof thestud iesepored i! thispaper we’eto idenlit po$iblei.f€ctivehossmd roures ofHcVviusoFrralrnission The aim of the lint *petinent wa to derermine whether geyheadedFuir ba$ (,”,/,papriotephal*) avtd Ee infeaed wnh HeU dcvelop be a source ofinfection for horse.

Th€ secondqperimenr inv$tigated a nodei for m ou$reak of HeV in a groupofsrablcd ho4es, by invati9ling if horses are susceprible ro notr-parenreral rours ofinfecdon,by identit/ing .he likely routes of virusexcrerion from horses.

And by derernining wherherinfeaed horeo couid generate aerosokcapable ot nansoiftingdisae ro otherhone andcars.

The thnd *perimert irvesdgared wheiherinfecredcarscould rlinsmir rhe Materiats and methods Anitul bd”dline drd nnrobiakgbal scfuit! Nl ‘dtuplins minats werehoucd i.

A high hicrcbiological eturiir dimal ooh at phlricl conrainme!! lcvcl4.

The remp€alure in rhe rcon w* mainhired at 22’C wnh 15 changes ofd! p€r hour The humidiryin the roon ws not.onmllcd, but maintained besan 40 dd 60%.

Vhen hmdling inoculated animals, the shfi wole tuUyen@psulated suitswith brarhing appr€.us.’r Clinicalsignsnonkocd ro indi.{e dis*e we.ed.hy..dia, hlperpnoe rnd abnormal behaviouch*acterised by dullnes or lerhargr,md morqia.

A hca( GIe noditor (Dynafii Pulscfionic, HRM10) wr modified for usewith horses, as pdlpation or ascukarion wasnot possible whilc we&hg rhe encapsularcd suib.

Vhen sdere clinicat disee dwelopedin the ( 1.

I mg/k), .nd hoss, theywec hevily sed*edwirh r7iazine killed usinga caprive bol! sun ald exsarsuinaiio!.

Rruit bas md .{s werekiliedby €rbon dioxidegs inhilalion,beeuse oncelhese species we( atrected by HeV the ce ofpuentenl injedios qs forbidden tor peGonal safety so6.

SpVirclogiel, serolosiol, hisrcpathological .nd EIISA Enzymerinked immunosorbent assay HorseEdish peroxida* Phosphate bui,Gredsaline SNT setus ndltElisalion lest 50’/.lissue cultlre inedive d6e 813 innunohisrochemical exanimrion i:ron all experimenral wec animals.\1rus isolations*rd dlr.dons ard serolo8ical!es!s performed at physical containnent Level4 in a I’lexiblefilm vitul i,ocalaft -‘the I],ev h @ulum ued throughott thn nud) Ms p*s€e 5 in Vero cells dd conuined 1067TCID5o p{ t0 FL.

Ihis rm the sm€ stock used by Westbun a alrlr: and had beenshown to be hishry prh%enic in d6 and guina wd used ro challelge pigs.

A standard dose of 50,000 ‘ICID, all oimah.

Exept hons 6, 7, 8 md 9, for which t06 TCID’ wo ued, as rhey werc challenged b1′ non’garenteralroutes.

Lt wr not hown whether horse were susceptible ro nor’ p.renreral ihfecrion, so a challcnge dose was cnosen to nuimne the chanccof infection. (pecrden.

Fo, \iru: Lolauor wc..

Labo.ato,l rudn” liozen at-80’C.

Swabs were blen itrb olhcted oeptielly and med E.on , r ning t Bs A.

Penr < illn. ' nepr om y . 'n -,irn.po l ud genBmicin.

Kolalion and tit4tion we.e undertaten rn Vero’cell cultue * de*ribed b) Murhy et rl.r Sahpls were passag.d Nice bclbre being regarded oegative.

Serum umplts mre te*ed for specific neutralising artibody by SNT and by an indirect BLISA.

For EI.ISA, 96-qclt microtirre pl.tes were coared wirh a l:400 dilotion of Hev-infected cell lysate.

Rkt ,q; d ilrred r:1 00 r nd “dded r o he c or ed m c r o t t r r c “e re plates.

Bouf,d antibodies werc detected by the addition ofa l:10,000 dilurion of prorein G HRPO conjugarc (iCN Biomedicah) followed by ret.ameih)’l benzidine as the ch.o’ mogeo.Sed givinsan densiryofgrertcr rho 0.2 above posi rhe mcan ofeighi conjugatcconfol wells were considered tive- sanpl€s for histologic,l analysis were fired in l0% butreredneutrJ fornalin and pahfin embeddedsed’ons we€ preparedby routinc methods and $ained wnh haehatorylin and€osin.ln additior, seciotu wereepmined fbr HeV aodgen using indirect immunope.oxidasc stairirs as prwioudl.

Describedby Munay er al’ btrt lsing a high &.e, polyclonll nbbit serum to inactivrted HeV Exptrinent t: Expdituektdt i,JLtion of f”t bat’ dsd dtt,npEd b4Mision to hoet El*en male gy-hcaded lrun BaB (P ?olloc?hzl@ wde obta;red Fom a spdve colony at the Univers;r/ of Queensldd and befoF inocuiation we€ shown to be neg:dve for antibody agains! HeV by SNT Uri.s the std’ Injc,aon dld do.e lour b’ *.- in&ulred by {h’u@eda over tfic thow and uother four ba* were inoculated by the :n,| ‘naali orJ rcu’e. -fhe ,en”in:nE ,h.e hr, were nor Inoculated but emained in conhd.

The bars werc eged o follows: cage I one subclrareoss irj€ction lnd onc in-co.tact bat, age 2 – onc intenoal/oral incrJaed bat, case 3 – Mo subcu6neus injecdon bats, cage4 so inhnasal/o.,1 inocdated bats, ese 5 – one subcutoeoG iniection and one r.

Cont.ct bar, and cage 6 – one inr€nasal/oral inoculaied and ore inAll bar casesst.e suspendedabout l.t m above rhe floor with dree eees adircenr ro qch oftrc ho6$, one gelding and n arc.

Which rere dc ‘ ; gnJ ‘ edho ‘ e I dd hor .

E 2 , c t p c c “ne tively.

This pcitioring of the qg$ rllowed for ihe exposure of ihe horsesto e*pied air ind urine €rcm the fruit bas.

Urine ud hecs of rhe bas were collected daily frcm the @Uection trays beneath &e bat cagesand wer€ added to ihe drinkiog warer ofrhe adiacenr ho6e- Swabsofrhis Nate.iar w€le lrso collectedfor virus isolation.

The hores and baa were exoined daily for the dcvelogment of clinical diseae.

Three weeks Pl, the bats were killed and necropsied.

All bzts were bled ar 8 dals PI dd et rerminaflon, 2t rnd 22 days l’I, and *rum ie$ed by SNT md ELISA.

Ai nampsy, smples ollrcted fo! vnus isclation wee lung, kidncy, splen, mesenrcric lymph oodc, liver, colon, urine.

Rdar sab ard braitr.

Specim€ns for h’sropaihologial analFjs .nd imftunopebxidar eehinddons frch bats includ€d bEin, sdinry sland, heart, luns, kidney,skeleralmuscle,stomach,smarl inre*ine, colon, liver, kidnev, spleen.

Bldder, skin, €ye ald adrenalgland.

The @o holsesweE bted for s.rolosicd ermi.

Nation 2 weelts alier eu$aoia of the nuit bas.

At thc conclusion ofrhe trial.

Rhc wo horss, as a test o{ suc€pribility, weE inoculatedbv subcu€neousiojecrionwnh a d ‘r r c a l .

I s n ‘”e ,e r”ndtuddo.coiHev Bod) (empentur “-d monitored daily, but heart Er€ ws not monitored in thse hoses.

As a Gsi of vi.ulence of the vitus inoc um Mo fema-le at one end of ats wcrc houed individully in wne n6h cages with rhe the aninal roon and wee ino.utared subcutaneously E ?dnht 2: E petinentzl i”fecnotu ofho6e’ ard atteqpt d ttawnniox to haret awl cax – Two geldinss, deslgnated horses 3 and 5, were inoculatedby sukuhneous injetion of50,000 ‘ICID5o of HeV Two nrarcsdcsignarcdho6es 7 and 9 werc inoculated by n intranoal srnb and l1ushof5 mL oisuperq 1 r r .

N r . .

O n r r i n : n gr r o u l o f l 0 ‘ l a l D , r H c v H o r e \ t a r d werc inoculatedon d.y l, while ho.s.s 5 md 7 were inocuiarcd – d a ) \ 1 , , e , .r o e i e n t l r h e p c d o d o f c / p o , u r o i r h e i l o n Gc’ animalsto infectcd animals.Horses4, 6 md 8 and ox catswere hoculatcd and in-contact horscs w€r€ housed in ahonai€ stalls (Iigure l).

Lniti,lry, es wde housed i^dividualy in suspendedcageson the ouride ofrhe horse salls, above tne Fcedsough, i.

Odd ro mdndn contac vith ioiict€d ho$es.

Ar inocularedho6es we.e euthan*ed, cais were pl,.ed wirh .

Of rwo sccondin-contact qt, ultimalely esultilg in thlec cages All animats were obsryed dairy and doniioicd at orh{ dds bv yidco suNeillance for rhe derelopmclt of clinical sjgns.

Swabslbr virus isolarion werc taken daily from urine on the floor of the horse salls md aft€r nedurenent the cdar rhe! monfterws dipped in rdspoii ftedia $ a t$r off,ecaldde iion.

NNI Mbs wcre ake n ftom inoculated ho6es that had m €lmted bod/ Ehp.ature.

Ho6$ tha! dcvdoped sde!€ clin’cal diseoe rvere euthuased witnin 36 h ofd€velopiry clinical sisns, and suNiving hoks ai 21 daF Pl.

All borss were n€cropsicd md sera llom the hores was rcsted by SNT and ELISA fbr HeV aniibody, .!d .u ho.seswerc necrcpsied.The in-contact cats md hono were bled 3 weeks after euthoasi: of the lst inoculated hoc.The etr were killed od lecropsiedat this timc md smpls .aken for histoFthologiol &rlysis ad vilal’ and dtt,n?t..! E*p.indt i”f..tio, 3: E peia.atdl mnnission to hores -Ihe three sednegatjve ho4es.

Ail dares fiom rbc previols lrial (hoses 4, 6 ard 8), wele used.Twelve ftmale cars were inrodu.ed into the rood, 37 days alier rhe the lst comhen.enent ofthe prdious tdal ed 16 days ‘1ier infeded hoe hed been neclopsied.

The ca$ wele inodlated by the onl route which had been shom to initiate inhction pr€viouvrr wkh a doseof50,000TCID,o HeV The/ were hosed i! .rsd scpended on rhe nils of rhe thre horse $rk (Figure 2).

Housing was in pairs excep! for two cases which each Two qs were eudanased each dr)’ !I for 5 da,vs(rigure 2) as put of a sepamte expetinenr to stltdy thc pathogenesis of ihe 814 ld Y,r/ vol76, No 12, De.embd 1998 TE– l f I I ‘l* 1 b l * r lE I gJ I I q*l gE 2, FlglB 1.

Plan oathe anih.l reoh al 0re beginnlng 6r a4erhdt showlng the posidon ol inocul.led and ln.conraci ho6es and roure, thrcc bab were sercpositive, lated by the inrooal/oral Mo with lower tirs of ]r5 and one with 1:10.

The *rum of the fourth bat in this group was cytoloxic and to result w$ Fcorded.

A1l eigbt inoculared bats were setopositire by ELISA All in-@.!ac! b.ts wee sercnegaiit by SNT and BLISA No s.o$ lesions were obscrued ar necrops/ in an) bar’ ldions and posidve immu.osaining for HeV wer€ @nfined ro the Mo fruh bats wirh arribody titres of l:40 and l:80.

The lesiors wcrc more numerousin fic bat wiih d anribody titre of 1:80 ard Rre charuaerisedby innammation of blood vessels,pa.ticulffly rrieries of the mesentery and alimen tary traci There was enher Fark d to modeetc lymPhocyric i a f i l ‘, *, o n o l r a – J n o r d r e n L l r u r n c p e n ‘^ , u L , u . .u e .

O ‘ fibrinoid degeneration of the vesselwall fith infiltration of losidve immuno$aining fo.

Lymphocltes and macrophasesHeV .nligen ws evident in the endothelial cells and snooih e o f r h e r ” t a m e d i a .\ i m i l r r b u r l e s i – , e r ‘e i m n ,u n o ru tainirg rvas evident in the vascul.r tissue of ihe kidnq’ md spleen,lymph \nruwu notisolated fron lung, kidney,Liver, aode, colon, u.ine, ectal sw.b or b.ain ofany fruir bat alicr oo pa$ages i! Vero ceus.

Swabs ofurine md faeces ftom the collection trap urder the froir bats werc atl ncgativf The so \ cats inoculaed deleloped clioica.l diseae ar 5 ind 9 das PI and HeV ws isolaled fton their lung, kidrey dd spieen.

Of The so in-.on@t ho$6, bled 2 weekr ali€r euthanasia the fruit bas, were seronegative by sNT and ELISA.

Nner chall e n 8 .

H / . , b c u m n c o u .

I n i e .

R i o n , \ s b e & m e .

N o ! .

I .

P\,fl \ (39.2″C and 39-7″C) wnh incrded espintory iate at 5 ud 6 dals P1, rcspeoively, and were euthansed 24 houn later HeV w,s isolated from the lung, kidney, spleen and utine ofeach ho^e md ftoft rhe satiE ofho& i.

No yirus was isolatedftom lyoph !ods, btood, faec6 or naral ca Y of bml’, prsapulf enher (Table1).

Inrtcno, of baA zsd attenPed E ?dnenr 2: E Vtih.s,ol ttax:n;ssion ra hor:es aad taf – Thie€ of lhe four inocuiatcd wi$ hoss (hos$ 3, 7 and 9) show€dclinical signsconsisrent and The results of subcutaneous infedion by Hf!:’a inrdaal/oral inculation of ho6s clrllengcd wirh HeV and rhe clinic”] outcome of the in-@ntd hors is summdised in Table l.

Ho6e 3 had an inderscd respi.atory atc, a body rempenrure ofl9.2″C ar 5 days PI and be@ne progresivell leliegic and anorexic until day 7 Pl when lhc hoB€ bed a o i , 1 0 {‘C .

R h d , , r ‘e o f a j b e , ” P € , d ,h u r e , ‘m p e r a , u , c nyoclooic twitch .nd bDb of the head, od was rhen eufiansed.

HoneTws letharsic,shwed d hdeascd rdPiration nk $d a body iempc’atureof40.4″C,7 di)r Pl, md wa euthanded 24 h larer On day 8 PI, 9 developed a Gmperaiur€ of39 9″C which lose !o 41.3’C for the next 2 days and returned to 37.8″C on da)’ t 3.

Duing this ftbrile period the hor* appered an ince$ed and shaliowr€spi.rtion and d lisdes uddutl,h ircrsed heel mte ofaround 65 bcar5p€r m;nurcfios 9 to 10 days Pl.

Thn horse had Hev’sp€cific neuRlisins dtibody by SNl’ar a tift of l.t l? a! 21 dals Pl, whe! it wa euthan4edthcrc were no ssss lsiotu at the tine ofeutharsi..

Horse 5 did noi ddetop ciini.J dis€ae ot seroonvert.

Lt was killed at \4tu wls notislaed frcm 2l daysPI dd had no gro$ iesions.

On necropsy, hos* 3 and 7 had an ad”anced diladon of thc lymphatic of the anterior venrral lobes of the lung.

Thc predominanr histologicdlfindings ir both hores were intcar’ ot etpe.iDenr3, Figure2.

Plrn ol the anihal bm at tne beglhnlhg 6i3 and i.4onl.ct ho6.3.

Showlng th6 poslllonof inoculal€d vnh ihe rldy Pl on whlch lhe PM= necopsy lable, Ct = €t @Oe cab ih that cagereF n@Dp3i€.[ so caBwerealowed!o discase in ftis sp€ci6.lhe remainnrg sages of dis6c and w€rethen erthanised.

Dcvelop advanced .

B ra i n .b l m d b u n etu to – m e *nteti c L rng.

Li, lno. .

Plc er conjurd;vd, iasara.d lymph rode, urine.tachcal,brcnchial, oral s%bs ivere cxlmined by virus isolalion.

Blood was collectedfton all c$ ar .ec’opsy for sNr &d EUSA.

Hos6 wereobseneddaily for fie developnentof cliniel disae.

Ho6es(hors ofthe tial, lhe nvo suryivins A! rhcconclosion negative by SNT and 6 md 8), werefound !o bc serclogicallr I TCID5o HeV inoculai;on of S EUSA andwerechalhng€d b Hose 6 was challeng€d usingdifferenrroutesof challenge.

Orallyby emptyingthe slriiec diEcdy inro tle mouth while Hosd 6 dnd 8 br innn*al swabbins.

Hose 8 w$ chall.nsed 8 wasbledat 8 and werebledar 2, 4 and 6 daysfI, md horse flom both at necropsy l0 daysPl.

Blood w6 aho collecred horses.

Virus! wd anempedftom the buffv cot and Results Ettc/in.”‘ t: Ex?einehtdt inli io, of fun bax and attenp’d t/ansni”ion .o hor:er- No fruit barsir this experi nent showed clinicaldissse during the 3 weekobscrvanon period.

Ar 8 d.ys Pl, no antibodiaag.itst HeV weredetecred neuualkingantiio $e serum ofoy bar At 3 qcek ll, serum bodics at tiiresof 1:40 dd l;80 werefoundin Mo of !h.

Fou’ b:r, irorul:’ed by rukotrncou, ‘1ie.,:on.ln the Crcupin”, u lut Ydl Vot76, No 12,Ds.nber 1998 8r5 T.bro 1, i..ult of.u6.ularu ch.rr.n!€d $|h rl.v .nd ih.

Cllnlol our.@ of h.6Lci Hce numbe.

Rode ol cbllens€ R*ult h.Es. Clini€ldb*so, €dhan.s.d 7 d:Ys Pl Noml .’rhdAs€d 21 daF Pr c ni€ dE€a$ eumhas€d 3 dals Pl c ink3r d sa* b€lwen d6y 3 and 13 asdesc.ibcd tiei pneunoniaand s@ere uscul! deg€neradon p.cviouslyin acutc HeV in hores.’i The v6cula! l.rions muscle, alimenoccurred in ihe lungs,kidneys, herrt, skeletal tary tract, spleen,lynph nods, biaddcr and neninges.

Posidve Numerousblood veselsand sFq’t’al celb showed imnunosainidg for HeY ll addition. 3 had mild l)’nphoclaicneningiis wnh pos;tive inftunostaidingin tle leptomeninges and in the blood ve$elsofthe subanchoo’d space ud boin puenchymaIn horse 9, euthalsed tt day2l PI, thele {e!e chronic lcsionsin the brain, kidney,spleeo.

Rlincntary tnct, moinges, skeletl nurle hqn,lung, m6en’ tery dd nsal mucosa, chanctcrird hy markedperiafterial mononucls cell infiltmtion.

No HeV dtige. $d derfft€d b).

Imhunoperoxjdae $ai!ingVias w$ ielaied flon a nnge of tisu$ of the Mo adtely infecied horses.horses3 and 7 (Table 2).

And fton the urinc aid oralovitr of hose 3.Viru rits we hidd id thekjdnev than ir tle lung, md weremuch lo*r in odq orsm.

In lorse 9, the ho6.

That rccovecd md wd kjued ar ?l daF Pl, viru m iolated liom only one tisue, the spl€r.

Vnus *s rot koialod fron laal or mat smbs of hors ot of nom urine sabs ofde floor ofrhe hoNe stalls.At necrcps), rhe inodJar€d hoGs, virus wa not isolaredfiom tne rctun, conjunctiu, naal €viry of racha (Table2).

In-ortact horss 4, 6.

Od 8 dd e6 wererronegarive by SNT and by ELISA.

The @tswere killed and naropsied.

No virus re isolatedFon feh tisu* and thee wete no grcssor E ?titu.nt 3: E ?otmntal i,f.aio, of.aB r’nd ntt.n?t.d ndhttui!’io” of HeV to hory – Since inoculated cats wffc not all eurhanased in a sequential rime{ouse expelimenr, Clirical signs of op.!-mouth developed clinicl disease.

Breathingdd dyspnoa wcre obscflcd onh in the lN eo ois to bc euthan*cd, on 7 ud 10 dars Pl Bpectivel)a Th.

Gross ir lhe atr€.tedcas id lhis and hisrological lesions obse.vcd with rharsn in peiou studies.r’,r’?rt sudy wereconsisrenr \4ms u isolatedfrom lun6 kidre,v,spltrn, biood md urire of rhe fou.


HeV tircs werc hishesrin the kidley ai l0?5 at TCID’, Virus6 atso islared from the u.ineoft}!ft cars nft6ps)’, with viral dtr6 of l0r to l0r5 TCID’..

Two €6 that had HeV ;n thci! uriae werc adhcnr .o ho6e 4.

HeV x6 not isolatedlioh faes, nsal, oal or trachealvabs frcn any of of Ho.se4 becde deprdsed and had a body tempenture 19.4″C,atr inc!6ed h€anhte of62 bas per ninute, ed @ ircr*ed ldpintory eie 6 dals after lot contactwith the l*t ideulaied cat, ftom wasiolst€d fon rhe u.ioe.

Thse sigru rcF presot ir tfie hors 46 daF after apcure to hoe 3, a reielbourirt inl€ctedhoBe.and 53 daysaft..

The 8 r6 b), SNl’lnd sranofexpeihenr 2.

Horse4 wasseroncgaiive ELISA 4 daysbeforeclinical diszue dedoped.

Over the next fewdqs.

I6 bodyEhpehture6 40.t”C,40.6’C and19.8″C.

The hose had a slighr nel di{hdge, dd it wasletharyicrd anorexic.

En eutha*ed 4 dals afte! rhe inilial tempe€ru’e riselhe hose had seroconvc.€d.nd w6 pcidve by ELISA ud by sNT wirh d rite of 1:2.

On recropsythe lungs wcre Iynph nod.s w.rc monled, and colonic and mesenteric congcstcd.Histologicall* rhe lsiotr in this ho6e wer.

Sinild rhot (en ia i n ho,{ ( J .nd 7 rnd cons$enr ‘ o rhose ” i ,h qp€rinental and :ddi6€s ofacure HeV disrue,la prfliou in lhe dolally thee ws non{uppuEli’e seninsoenephalitis rcofof the oedulla.

HeV wa ished 6on blood, uline, spleen ud kidney (Tabl€2).

S*en day* afterlut exposure ro ioqlated cars,hoses 6 md nesadve by SNI and ELISA Afie! onl 8 wercserologically inocuiation, hose 6 dryeloped a body renpe€ulc of39.7’C and had a heart ste of 74 bea$ p.r minute on day 7 lIT*nry-four hours latcr, tlc lorse had a hean rate of 90 beats per dinute, a Bpiraiory rareof50 brcrns per minote,.d a bod/ iempenrurc of39.8’C, and ms cuthanaed.

On rccrossy the hoce showedgtu md histologicallsions consi$ent wnh lrom i varier)’of acrreHeV infeciion,ra and virusm isolared 2).

Virus wasisotdtcd lion its bulry tissues and urinc (Table oat on dalc 4 and 6 PI, but not at dry 2 PI or at rhe tinc of eu$arsia.

Ho$e 8, ino.ulaad by a n*al mb.

Had no d*elopedclinical diseasc by ]i dals Pl ud Ms euthanacd.on ndrcpt.

The ho6e wd s€lodegatnt,no vtus m isolatedfrom in .h€ t’ssues blood or rissu$ add no lsions wereobserved (Table2).

Discussion vhen chalensed with HeV six ofcight Fuit b{s erccon’ vened without daeloping obviousdiniel signs.The only vo frtrit bls witl scula.

Tesions wereth€ bas wiih the hishes antibodylitEs and $ mtibodr rite appea$ro be .oftlat€d Anlibodig inra dorded in the six wnh the swerity ofdisease.

Ie consistentwith rn.s eord€d fru;r haa that seroconvened in ficld studies(P Selkk personalcommu .ation).

The vascular Iesions that were obsened i.

Lwo frun b!6 were pigswith obseaedin guin€a morphologigllysimilarto those HeV inf€criortrt bur ditTe.enrfion rhose describedi! to be reso hors.’a Fruit bac haveprwiously beensuggested of !ht “rud) i’e.onrhten, rci- hor ofHefr0 od rhe6ndings wirh rhis h/poth$is.

Th€ hilure !o ; fron the !tu patholosic,l chrngesand positive b*s that dcmonstrared of rhe immuneresgonse immunoslaining is not surprisingThe rh* be$ suggds rhat viablcvrus had beenelim;natcd.

Antigenof positivcimmuro$aining is likely to be delecting udviable’virus, tbr .xdple nu.lsc+sid prorein,Blhe! rhar Experidenrs I and Z indisre that HeV is nor higHy infb.In dper tios, at les under thecooditio’softhe exp.rime.ts.

Uine and h€es of the infsted imcnt l, rqpiqtory ‘€rosols, adjacent ro the fruir bats,dd aerosols fron rheir oecropsies how, failed to initiate ilfectior, fd therests !o rp@d liom pe.iod.

In fruir bat io fruit blt ovc. .he 3 week pclirf.ctio! *ery appdent opportunityfoi aerosol experinent2, despite fiere 6 no (rtumrsion 6om ho^6 to the in-onun ‘prrd.

Fton rhe hors or ca:s.This wu corsislentwnh the resolrs vi.u isolaliotu.

HeV m nor isolaed ftom the naial caviry or horses.

Lurthermore,one horseinoculatcd uchea of infected .46t val Vol76, fio t2.

D€ambe. 1998 Dlss (7 d) orw* (53d) Dis** (3 d)

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