Hamada. F. K. et af•.•• ISSN 1110·7219 III
STUDIES ON PESTIVIRUS INFECTION IN SHEEP FLOCK (BORDER DISEASE)
'Hamada. F. K.; Iman. K. A. and Zeidan. S. M.
DepL (I( Anim Med, Fnc Vel Med, z.'1g.tUp. UnivcrslLy. fit:nlm I:iraTll:h. EJ{Vpl'"
Veterinmy scnnn and vaccitle H(,Se ABSTRACT 7he current investigation was per:Ionned on sheep ,flock of dWerenl ages, sultefln,q from pfOblems oj a hafry coat muscular trembles, aOOr/ion. stillbirth. weak and Jrafl iambs, barren ewes, nasal diScharges, intermittent diarrhea andJew deaths. 'Ille clinical cliterla suggested that. lhLs field problem aHrihu/(>d 10 a pesfiuin1s h IecHon (Border disease virUS. m)V), The virologicaL and serological analusis pfOl.'ed tile detection qf BDV and spec(fi.c neulmUzin1 antlb Epidemiological and laboratory inuf'sligatkm in this study ckw'/iJ indicates Hwl 1J1!f JaIo calves iJifeded with aVD virus constitute a source oj Injection Jor sheep. tl'hicll rt'" suits In border disease. Also epidemiological data ret,'culed l/tal the nli)rbidiLl) «f Hw disease was higher in newborn lambs (32%} Jol!OH'f.Yi. by oldcr lambs (28.0iJ4JJ (HId adults (5.9%L The bar(erioiogl.col examination delec/ed snltrronella species am! E·Co!i as fnlercurrenf i1lfedion (0 B[)V due to illtmW1QSuppn!s!>(ve action oJ tile viru."O. INTRODUCTION lJcpending on the host origin Pestiviruses \ 'ailed bovine virus diarrhea virus (fWD) hog dlOle· ra 'vIrus (Hey) and border disease virus (ilD\ 1 in (:attlc, swine and sheel) respectively, Cross in fection between species nm be achieved cX1W)imentaUy and h;):s b('en uemonstrated in Held con ditions (CoUett et al" 19B8~ Franekl et at.• 1991 and Nettleton and Entrlean.. 1995). BVlJ and ED virus;:s have a comm:x: origin and those f'hangcs lU tit(' cpitope. t'xpn:'ssion 1I1lgh! ,lriSC' Mansoura. Vet. Med. J. (111 • 129) VoL N. No.1. 2002 Hamada. F. K. et at ... 112 during passage in different speeies (Carlsson and Belak•• 1994). BVDV handkaps basic 11> search because it is a frequent contaminant of cultured cells. This contamination most frequent~ ly is the result o( ubing fetal bovine serum, derived (rom BVDV infected animals, as a cell culture supplement (Nettleton and Entrican •• 1995l An assessment of thc economlc losses due to infertility, abortion. nconatallosses and low {'ar cass weight mdlcate that an outbreak of border disease can result in a potenUal redudlnH of in· come in excess o( 21Y}f, (Sharp and RaWSOD•• 1986). Sheep to sheep contact is considered to be the principle way in which BOV is spread (Nettle ton •• 1988) but cattle persistently Infcc1.ed \.\fith BVD and in dose contact with sheep are an im portant source of strains of virus capable of causing oorder dIsease (Carlsson •• 1991). The spread of pesUvituses can occur among both cattle and shcc.p where sheep and cattle are housed together (NUsen and Slettbakk., 1995). It is dear that, lhe grcat ma.lorHy of BOV infections are subcllnlc:al (Hungerford .• (990) and the pathogenesis of BDV Is govemed by vanous factors associated 'With the immunological stat· us of the ewe and the age of the fetus at the time of infection (Hu$Sin and Waldehlwet .. 1994). BOV Can cross thc placenta and establish a persistcnt infection in lhe animals infC(',ied tn ulero (Caffrey et at., 1997). Infection of ewe with non cytopaUlic biotype of BOV during early pre~· nancy can result In birth of lambs persistently Infected 'With the virus (Loken .• 1995). These Jambs are apparently healthy and can develop and reacb malurlty and can be bref'd and birth of perststenUy infected Iambs and all these persistently infected animals shedding virus in Ill(' sali va, urine and feces for the rest of their Itves (OebW'D and Ca1Jtnleel.. 1991 and Wolf and Butt ner., 1994). It is a well established that. BO 1$ characterized by variable clinical manifestation namely haIry fleece. lower birth weight. ncurologlcal dysrun(:tion, re BDV infection before the development of the immune system (60·85 days of gestation} may re· suIt in bIrth of viremic and perslstently infected lambs without antibody in precolostral sera (Barlow et aI.• 1983). Fetuses that survive early intrauterine infedion with pestivlms win hom virus~positive and antibody negative while those. infected In later gestation Win be anti- Alansoura~ Vet. Aled. J. Vot. lV. No.1. 2002 Hamodn. F. K. et at. .. 113 body-positive and virus negative. these persistently infected animals f'X('I"f'ie tht' virus in nasal seen:Uons and saliva (Nettleton and Entrlcan .• lOOtS), aVD virus has been shuwn tu induee profound tmmunosuppression whH::h may be con" tribute to the pathogencity of the virus !L"elr and perml.! secondary inredion 1u ('nduf';. The v!nts has been implicated in 111(' depression 01 T-Lymphocytc function. antibody production and the function or mononuclear phagoeytcs and neutruphils (Markham and Rammaraite: .• 1985). Serology Is Important In monitoring the introdudion of BDV In a flock and lor tht' rctrospt'f..' Bve Investigation of new outbreaks of BD in sheep flocks, n,c prest'ncc ur absenet" of ilntlbodies in sera or lambs depends on the immunolOgical status of the eWf'S and the age of tl1(' fetus at the time of exposure to the vtrus {Husain and Woldebiwet•• 1994}. The seropositive sheep La peslt virus occurred due to Infection of the retus after R5 d.-1Ys of ge-stahon or to post-natal inf{"dion and these animals Is very 1mpMtant in the epidemlop!ogy of pcsUvlrus as trdnsmH 11lfN'thm to cattle and exaggerated problem of I}VD espe.:iaUy there is no vaccination program adopled In Egypt to control of pestivirus infeenon In sheep and goats (Fouila .• 1999), The antigenie relationships and physieal and biological Similarities between BVD and BOV had led to the assumption that both viruses belong to one species of the genus. pcsttvirus (Oone et aI •• ]988) and liDV and BVDV indistln~ulshable in vllm (Harkness and Vanta!s .. 1982). Concurrent infection with salmonella and BVDV would 0(' ('"xpc("ted to caus!' 1110rc SCV{'I"('· dis ease than infection wUh either agent atone owing to the imnmnosuppresslve cllecls of llynV in rectIon (Wray and Roeder., 1987 and Penny et al., 1996), It was emphasized that. in the control of border di.&ease in shet'p and bovine diarrhea virus Infection In cattle, it Is important to mInimIze the rtsk or transmiSSion of these related pe!Stlvlrus, es between lhe two spt'des (CarlslIOQ and Belak•• 1994). Few published lit('rature data eonceming pt',&tMrus Inre.Hun or border dist'a~(' ln Egyptiall sheep (Shehata et al.. 1989: Baz •• 1992; Zagbawa•• 1998 and Abd EL-Haklm., 1999: Salem et ai.• 2000 and Azzam .• 2(02). The present study aimed to look for a problem of abortion, stillbirth, barren ewes. ilI-thrjft lambs, respiratory distress and Intermittent dlarrlw.a among sheep of dlffer('nt ages in private sheep Hock with particular emphaSiS to dedare lhe posslhle ('ausetsj from laboratory In ...t'"sU~ tions, rnonitortng and descnption of clinical pi('turc, assessment of some epidemiolOgical data. declari:lg the incidenee of intercurrent inrection and the suggested lint'S of control of lht' prob lem. Mansoura. Vet. Med. J. Vol. IV. No.1, 2002 Hamod«+ F. K. e:t al .... 114 MATERlAL AND METHODS 1 ~ Animals and history of the nock : ThIs study was performed on a tolal of 243 cross breed sheep uf different ages rrabl~ O. they were seiected from a private farm at Shari 2~ Reference virus and antiserum: Reference avo virus and antisera (strain Orcgon C24 vi rus which Is clo8ely related to border disease virus) oblained from VeLSerum & VaC('\ne Res. lnstJtute. Abuassia.. Cairo. There Is no available reference antiserunl and antigen for BDV in Egypt, so we used BVD virus and spccHk antiserum (serolugically related to BVDV, and often cross neutraUze each other). 3- Cell c:ulture : Madln-darby bovtne kidney cell eulture (MOBK), kindly obtained from VeLSerum & Van:-ine Res. Institute. Aubassia., Cairo, Egypt. It was used in detectinn ur the virus and tn serological identlf\caLion. 4· SampHng : TIle lIiVestlgated sampies were taken from clinically tit anim~ls {32j and clini cally healthy contact animals (55). these animals were sampled for twu Hmes with 3weaks intervals. Also contact fattening buffalo calves (251 were sampled, A- Recta! swabs: Two swaus were taken, the nrst one on transport llHXlta ror virus dd('c, lion and the second one nutrient broth for bacteriological exammahon. B· Nasal swaus: It was coUccted on transport media for virus detection C· Serum samples; These samples were taken from diseased and clInlcally nOnTIul conIne! newborns. older lambs and adult.s sheep (prec:olostra! serum obtained from 3, lambsl 10 detect pestivirus (BDvl and antibodies. Also serum samples collected from raUenin~ buffalo calves In contact W1th the investigated shcep flock. 5- Laboratory tnvest1gatlon: A- Virological examinatlon: The collected materials were prepared for Jnoculatinn toto MDBK cdl c.ultures and examined for cytopathic effed and the presence of pestlvlms was determined and cooflnned by indirect immunoflouTscence test {lFAn, neutraitza tlon and the lnterterenc(' tests according to Gilleapie et aI .• (lOO7). ElrBagotuy.• (1990) and Thabtl et al., (2002). Mansoura. Vet:. Med. J. Vol. IV. No.1. 2002 Hamada. F. K. et al•••• 115 B- Serology: Serum neutralization t<,st using reR-rence virus and antiserum wa .... applied (Rossiter and "eoette .• 1982). c- Bactenologlcal examination: RedLtI swabs inoculated inlo nutrient broth ami se\cllltc F broth were incubated for 18~24h at 370c. then a toopful rrom the grolAih inoculated 10 nutrient agar, blood agar. 5,S agar and McConky agar plaies and all plates incubated at 370c for 24~48h" TIle isolated bacteria were identified and eharack_rized from colony morphoiogy and bioehemical reaction (Quinn et aJ .• 1994). 6· Tbe:rapentic trial : The dlsease animals were treated symptomatically in addition to fluid therapy while those exhibiting intercurrent infection treated speeHically ac{'"oHhng: to tIlt' type of secondary invaders (Radostlts et aI .• 2000). RESULTS AND DISCUSSION Border disease (BD) could induce economical losses in nat ve flocks alone or in aSfS()ciatiol1 with other pathogens (Thabll et ai., 2002), Originally, pcstlvlruses werc desj~natcd as BVDV. Hey or BDV based on the host speCies from whieb til(" virus was isolaled. Beeause these VjruSC\i can cross species boundancs this nornenelature was pmbkmatic. Later allemp!S to group pel5t1- viruses have been based on serology and monoclonal antJbody 1,.rlndtng (Mettleton and entr!· can., 1995 and Martin and Aitken.. 2000). OD is a congenital virus disease of sheep, charac terized by barren ewes, abortion. stillbirths and birth of small weak l:-HlllJs and n wl.fi Regarding to virus detection and identification from serum samples and nasal or rcC'iat ~wahs (Table 2 & 3), the current virologieal studies declared detection of BDV (cytopathic and r1On cytopathic blotype) on MDBK cell culture, the pn:scncc of BD virus is established and IdcnUly by neutralitation and lntenerence test" and immunofluorescence teehmque. TIle above results w{'"re similar to theses previously reported by Terpstra.. (1981), Roedel' et aL. (1983) and Nettleton aad EOtrlCBO•• (1995). The detection of the virus (especially non qiopatbie blotypej at fmH and second eXamination In both clinically ill and clinically normal sheep deaIT'd the cxistcnce of perSistence infection ill the herd and these animals were responsible for the rCCUlTence of the infection in this flock. Mansoura. Vet. Med. J. Vol. IV. No.1, 2002 Hamada. F. K. et at ... U6 Such finding was Juslified by prior work (Woldehiwet and Nettleton•• 1991;Wolf and Buttner•• 1994; Loken•• 1995; Caffrey et aL.. 1997 and Abel EL-Haklm•• 1999] and iaJ11,l? number of pt'T slslently infected (PI) sheep were apparently healthy (Nettleton et aL. 1998). Virtually all PI ani mals are virus positive and antibody negative and a repeal vjrus positive sample 3-wet'k later will connon persistent infection and disllngulsh it from an acute infection (Nettleton and Entrl~ caD •• 1995). The detection of pestivirus {BVDV) and specific neutralizing antibodies from contact apparent- 1y healthy buffalo calves supported by the study of Carlseon.. (1991) who diagnosed border dis ease tn sheep caused by transmissIon of Vims from cattle persistently infeded with bovine virus diarrhea virus, Also Rad08tlts et at., (2000) published that, the major managemenl risk factoes In onset of BD arc introouCtion of persistently infected. shCf'p and rarely cattle In a susceptible sheep herd. Clinical signs described in this study was appearcd sporadically In form of less obVIOUS hairy. fleece and trembles. besIde abortion. stillblrth. v.reak and frail lambs. barren ewes. nasal dis charges and InH:rml:ttent diarrhea While other' animals exhibiting persistent or inapparent inft'c Unn and death of lew lambs. Such observation was suntlar to that observed by pctor work of many workers (Plant et al .• 1983; Barlow et at •• 1983; Shebata et at .• 1989; Rae., 1994; Net tleton et aL. 1998. Abd EL-Haklm., 1999; MarUn and AItken •• 2000; Rad08tits et aI.• 2000 and A..l\um 2002). The clinical Signs of the disease was marked on iambs than adults (Sawyer •• 1992} and on animaLs showing intercurrent infection {8anchla and Russo .• {l9901. Hairy fleece is due to predomlnance of primary meduUated hairs rather than Set"Ofltiary wool (Osburn and Cutruccl•• 1991). As well. lambs that survive may gradually lose thelr neff'e ab· tlormaUues and nervous Signs (woIdehlwet and Nettleton., 1991). Also perSistent infected ado~ lescent and adult sheep may appear normal as the tremors and fleece abnormalities abate with age (Swayet•• 19921. tn Egypt, maJortty of sheep breeds has coarse wools and the hahiness criterion of the wool Was less obvious on Infected shet'.p. This explanation is similar to that of. Orr and Barlow.• (1978) who reported that. tn coarije~neeeed breeds It may be djmcult to asS(;s the hairines.'3 of the bIrth eoats and Ott.• {1982} who mentioned that. the skin abnomlalmes are obvious in lhe very flne and medium fleeced breeds. In this respect Bonniwell et at., (19S?) reported an UOll~ sual outbreak of BD in lambs In whIch the typical signs of tremors aud atmormalitJes In n('Cce were oot present and the Signs were restricted to abortion and the birth of 8mali wcak lambs which weee found to be persistently Infected with BDV, The abortion occurred due to that, BDV cross the matemal vascular endothelium of the pIa- Mansoura. Vet. Med# J. Vol. IV. No.1. 2002 Hamada. F. K. et al.. .. 117 ccnla Within 10 days of jnjection resulting tn virus transfer from dam 10 oHspring or a placentilis that result in abortion or the pregnancy may be sustained If the placentiUs heal in aonut 25 days (Snowdon et at.. 1975). Table {4) revealed the morbIdity and mortaHty rates In the tnvestigated sheep HO('k. it cleared that the morbidity of the disease was ~)2, 28.6 and 5.9%; among newborn lambs, older lambs and adults re~pecUvely whlle the total morbidity within the herd was 13.2%. Mortaltty rate was 8 and 2.04% In newborns and older lambs while there was no mortality among adults and thc total mortality within the nock was 1.23%. The above rt'_lmlts wt're nearly similar 10 thut reported by MartIn and Aitken., (2000). From the epidemiological polnt of few. the tracing back to a possible odgin of tllt" disease, aU evidence provf'd that BDV infection might introdu~'ed to thf': invcsflgatlXi !lock due In {'{mtae! with infected fattening lmflalo calves and introduction of inapparent or persistent infcclcd shf't'p. As well, Clinical evtdcn.e proved that these cases appeared sporadicalJy at !rrt~gular i1ltervals. such crlterta mentioned previously by Radostlts et at., (2000) who publishc- Respecting the results ofSNT Hable 5}, the anulJodies were detccted in 18 animals and fhe- tit ers range was) /8: 1/64. The use of cytopathic strain of aVD virus in performing of Sl\1)" was doc umented previously by the work of Brockman et at .• (1988) who reported that avo virus CQuld be used to detect neutralizing antibodies to BOV In sheep sera and it had be!"n in usc for this purpose for several years. Neutralizing antibodies were df'teeted jn either clinically HI or in map· parently infected a.nImals, sueh finding nearly similar to that detcekd prf'viously (Ha1ez" 1973; Halun et aI •• 1989: Zaghawa" 1998 and Fouda •• 1999). While few infeded animals were' have no nCliiraHzing antibodies but have HLJV, this eontribuled to that persi~tcntly infected anilnals are immunocompromlsed or tolerant and have no antibodies (Sawyer et aI., 199il Tht' detection of S'N"T antibodies In precolosfral sera of sunlc lambs doeumented persistence Infe-euflll of lambs due to in·utcro Infection with the ED virm" Su('h finding was 1n accordanl'e to that presented by Nettleton and Entrlcan.• (1995) who found that fetusc,> that survive eariy In· trautcnne infeeUon with pestivlruse VIti] born virus positive and antihody negative while those Infeeted In later gestation will be antihody positive and virus negative. In lhis ('on text Vantata et ai., (1979) concluded that after in-utero infection of lambs with BOV, sOlll(, lamhs may be ap' parently healthy but continue to shed lhe virus persistently throughout lhrir life, The persistcnt ly lnfcC'ted apparently healthy lambs If kept for breeding they WiD continue to be a .source of Ill" Mansoura. Vet. Med. J. Vol. IV, No.1, 2002 Hamoda, F. K. et at.... 118 fectlon In successive breeding seasons horlwntally or vertically (Woldehlwet and Nettleton•• 1991). 'l1lt~ incldenf'e of Infected animals e.l(h~biUng intercurrent Infection as in tabli' (6) in which E Coli was detected as 25% in newborns and salnl0nella sp in 21.4 and 4040 ot' older lambs and adults respectively. The emergence of RalmoneIla or 1->Coll infef'ti(lns tn border disease Infecled. animaJs may attributed to a state of immune dysfunction accompanied pesUviruse:s InJ{~('.t!tll1 (Sawyer and Osbun:l •• 1989 and Nettleton and Entr1can .• 1995). Moreover Lamontagne et aI .• (1989} reported that thc most Important aspect of acute or chronic phases of BVDV or BDV Infection was the host's increased suscepUbt11ty to secondary bactenal or Viral tnfcx'Uon due to aiteration In the cellular immune responses inductng i.mmunodepn'sslon. [n thIs respc('t Hajtos et aI.• (1998) determined that a predisposing factor of Eperythrozoon ovis InfectIon In sht'"t'p was border discase/bov1nc diarrhea virus infection and Monies and Simp.on.• (l997} diagno...,rd a concurrent coeddlallufrction in lambs lnfected with border disease. TIle results provide substantial evidence thaL the specifIc and symptomatic thcrapy of cases of salmonellosis or eolibacUJosls as concurrent lnfeetion result in temporary recovery and relaps~ cs oculr aftcr that. '11)(' above results was supported by the view of Monie. and Slmpeon.• (1997) lvho suggested that, if lambs mth diarrhea fail to respond to treat.ment. the possil>lIlty of persistent pestivirus infection should be considered. There is a _prevailing assumption that per sistent infected animals tend to sUL'Cl.1mb to intercurrent infecttons. We are m need for organized control programs for vetennary services to be dTective in tenus of animal pesLivlruses preven tion. Perfect preparation and Virological examination of any vaccine produced and used in Egypt against pestlviruses where pcstivirus was isolated from live sheep pox vac,cjne anti v This study clearly indicates that under Egypllan condiUon buITalo c;tlves. which are perSis tently infected with 8VDV. constitute a source of Infection for sheep which rcsults In border dis ease. Further investigation should be done to dedarc the epidemiology and emnomlc impact of border disease in Egyptian sheep and goats and to Identify the extent of the relationship between border disease virus and bovine viral diarrhea-mucosal disease virus In Egypt trom molecular bi ology invesUgation. In CGnclualon. the maIn strategy advocated for lhe control of BD is identlflcatton and cuHmg of the pC'rsistent execrator sheep {the most important source of infection). avoid the exposure of susceptible pregnant ewes to SD virus, sp~lal conSiderations adopted in artlfic:lal breeding pro grams and reduce or minimize eont.act of sheep v.1th infected cattle or other source of infection wm playa crUCial role in the control of the disease. Also great caution during vae("ines prepara- Mansoura. Vet. Med. J. Vol. IV. No.1. 2002 Hamada. F. K. et al. ... JJ9 Hon especially living aHenui:lted one which are used in vac('inatinn of sheep lx'eaUtil' p('stivlms are notortous contaminants of laboratory cell cultures and cell culhm: medium and frequently contamLnants of modilled live virus vaccines produced on ovioe cells. We are jndff'd 10 vaccinate sheep and goats agalnst pestiVirus usln,g vaecine prepared from loeal SD v!ms to contwl the dis ease in Egypt. Ta ble (1): Age and numbers Qf examined animals. Animals age Numbers I I . Newbom lambs Less than I month I I Older lambs More than one month-less ~~ than il'ear · \ Adul~ 1-5 year 169 · , · . Total 243 I Table (2 : Results of virological im.'cstigation on first exami~lati(}l1 of tile flock. I p~_Wbl)r~ hlmb~ Otde~lll~~' T~ Adults , , Samples T.Nr. ' No r.f %, ' T.No- No (If % T,Nr. : NQ ,r % +V, • ,:y"'e +Y.:;-- --'-' '-_. N D N N D N D N D N D N [) N D N 0 IR,swabs 0 1 0 Uj , , 10 143 J J 12 3D , Ha.~wabB , 0 10 I 10 • 0 0 14 I 5 71 25 10 1 0 8 0 Serum 1 1 10 125 , , 20 2L4 6 4 24 40 I .~- IT~tlll_~r 1 1 2 10 25 7 , J5 42.~~ __1.L! J7 I ,,~'I2QJ R: Rectal N,,: Nas .. l N: Normal rutlmals 0: Dist\a$cd animals T:Tolal No; Number (3): Results or "'irological investigation un setond examinatiun or Ihe flock (Jwceks llost first examination). ·--~-·~··I Newborn IAmbs .~ -.~-. , -,-- I • SamJiles ; 'f,No :Nt'lof!% • TNo NoMj % , T.No 'So nr'% i I 'V' +Ve i . !+ve-L i . I']~I -.-~~ DIN N 0 N D:N D N D N D'N' D . R.swabs , 0 o , 2 1 llO 7.7 .I 2 12 ! 2 I . : 20 i Na.swabs ' 10 • 0 o 0 13 1 0 21 HI I 0 4 0 , Serum I I 10 FE16,1 ; 4 154 5 2 .2~ , 20 ~~2 Towl 1 1 " 11,7 1 7 ) i 35 23.1 9 4 30 14 R: Recta! Na: Nasal N: Nonna! animals D: Di~eascd animals T:Tollll ~o: Number Mansoura. Vet . .Aled. J. Vol. lV. No.1, 2002 Hamada. F. K. et al.... 120 TRble {4}: Mormdity nnd morCality rates orthe border disefts~ in tht in'llestig.ted sht'ep fioek. - , -- ~im. ~ of Morbidity rate. !Mortality ra.e 1, I .X~ mined • • 1 I N. % No J % ' . ~uum aI • Jambs I25 8 ,32 2 8 I iNewb~rn 1 I Older tambs 49 14 28 6 I : 2,04 1 1 . Adults : 169 . 10 5.9 i 0.0 0.0 I Tot.1 243 32 T13Z- 3 1.23 , ._. .~ Table (5): Results or serum neutralization test. -----_. • First examination I Second uamination Animals T.No Noof+Ve % TNo No of % I +v, IN D N D ,N D N D N D N '0 , Newborns 10 8' 0 2 0 25 10 6 I 1 10 16.7 Older lambs 20 14 I 3 5 21.4 20 13 2 2 10 15.4 Adults 25 10 3 2 12 20 25 10 3 2 12 20 1s-· -_. 1 Total , ) 32 4 .1 ' 7.3 21.9 55 29 '6 ,5 10.9 17.2 The titer of antibodies ranged from 1/8: l/64 "'one of three preclostral serum samples gives positive resuJts T.lbJe (6); Incidenc~o~the BD infecte_d animals exhibitin concurrent infection. Animals No. of Isolated bacteria ciinieally Type Number % ill animals INewb~m lambs 8 . ___.~.-,E",' ·::::coo:.h~· ___-+' ::.2 _ 25 IOlder lambs. ...-~.~-:Ic's"'a"'lm"'o"'n"'el"'la'-'S"'-t-"3 ___r' 2",1",,44 Adults ~ _ I 10 __..i Salmonella sp .L04e-__..L::4",O_ Fig (1): BD infected lambs showing hairy lI.ec. (less obvious), Mansour-a, Vet. Med. J. Vol. lV. No.1. 2002 Hamodt:. F. K, et 01. ... 121 Mansoura, Vet. Me-d. J, Vol. IV. No. I. 2002 Hamada. F. K. et al. ... 122 Fig, 2:: BIl inlN':lct! lamb shoWing diarrhea du{' to inkrcu!Tf'I1! 1nlt-diml with E·Cnli. Fig. 3: 13D infected lamb 5h()WllIg (lianiwH due to mt('rt'UlT('nl in fiT I 101\ w\111 Salmondl" 'ip. Mansoura. Vet. Med. J. Vol. IV. No.1. 2002 Hamoda. F. K. et al.. .. 123 REFERENCES Abd ElrHaklm, U. A. (1999) : Border disease of sheep in Egypt: J-cli.nlcai and diagnostic (imli ret~t enZ}'me linked irnmunosorbent assay and reversc·tranflcripiion polyrneraHc dmin reactIOn) studies, 5th ScLCong, Egyptian Socliy for cattle djsea~es, AssllJL. Ef!..ypt: HO- 85. AszaDl. R. (2002) : Sludies on the epiwotic prQrf:SS of pestlvlrus Infection in animals. 1'h. iJ. 111esis (Anim, Moo. Dept), .Fac,VeLMed. Benl-SueLCairo University. Barlow. R. 14,; Gardiner. A. C. and Nettleton, p, E. (19S3) : The pathology of spontaneous and experimental mucosal disease-like syndromc in sheep recovered from clinical border disease.J.Comp.Puthol .. 93: 451·461. aaz. T. t. (1992) : Bovine virus diarrhea-mucosal dist"3.se and bonIer disease in f<~!ZvpL Rev,$cLTcch., 11: 867·872, Bonnlwell. M. A.: Nettleton. P. F.; Gardlner, A. C.; Barlow, R. M. and GUmour. J. S. (1987) ; Border dist'a.<;e without nervous signs or fiee,ce ehanges.VeLf{<:>(:>. 120:246-249. Brockman. S. J.; WOOd, L.; Edwards. S. and. lIarknes8. J. W. (1988) ; Sekction of Dn appro prIate pestlvlrus strain for border disease serodiagnosis .Vel-Rec., 122:586-587 Caffrey. J. K.; Dudgean. A. M.; Donnelly. W. J.; Sheahan. B. J. and Atkins. G, J. (1997) : Morphometric analysis of growth rdardation In frtyal lambs following expt'!nwntal jn~ fedton of pregnant ewes with border disease virus.H.es.VeLSci .. G2:245·248, CIU'18son. U. (1991) ; Border disease in sheep Ull1S("(i by transmission o! vims from (·dlth' JltT' sist<:>ntly infected wUh hovlne vims diarrhea. VeL Ike., 12H{7]: 145-! 47. ClU'ldon. U. and Belak. K. (1994) : Rorder disease virus transmitted to sheep aIJd cattle by H persislently infected ewe: EI>idemioiog'l and control. A<:>ta Veterinarta Scandinavlm., :35 (I): 79-98_ Collett, M. S.; Anderson, D, K, and Ret.zel, E, (1988) : COIllp<"1tisons of the pest! bovine virus With the memuers of the [·'}avtvlridae.J.General Virol.. 69:2637-264a. Done, J. T.: TerbeekJ. S.: Riccbardson. C.; Hakness. J.: Sands. J.; Patterson. D. and Duffell. S. (1988) : Bovine virus dlarrbea-uHlcosal disease virus pathogcncHy for thc fetal {'aU folk11.1 ltl-Bagoury. G. F. {1990} ! Evalualion of some recent virological tcdmiques in the diagnosis or hovlne viral diarrhea-mucosal dise-ase complex (DVD/MD.Ph, D,Thesis. Fac, Vel. Meci. Zagazig University. Benha Branch. Mansoura. Vet. Aled. J. Vol. N, No.1, 2002 Hamada. F. K. et al••.. 124 Fouad. A. E. F. (I999) : Sero-survey on bovine herpesvirus-I, peste dcs peWs ruminanis and border dIscase viruses In sheep, MVSc Thesis (lnfeclinus diseas~ eSI_Fae .VeL Mcd.Alexandria University. Franckl. R.I.: Fanquets. C. M.; Knudson. D, L. and Brown. F. (1991) ~ Classifl<:ation and no· rnen('ialUre of viruses,Arch. Virul., 2:223·233, GWe.pte. J. H.; Bartholomew. P. T.: Thomson. R. G. and Mcentee. K. (1967) : TIlt' isolation of noncytopathic bOVJne vIrus diarrhea vIrus from two aborted boVlne fetus('s,Conwll VeL 62:564-571. Hafez. S. M. (1973) ; GeographIcal distribution of bovine viral dlarrhca·Mueusat disease (BVIJ MDl in Egypt as dctennlned by the detection of neutralizing anttbodJ("s in si1Cl"'P St'nl. J,EgypLVeLMed.Assuc., 3512J: 73·80 Hajtos. L~ Glovlts. R..; Erdos, A. and PolO. V. (199B) ; Oiagnosis of Eperythroz{)oll ov1s infec tion tn sheep in Hnngary, Preliminary communlCaHon Marorar 01latoxvosok Lapju" 120 (11): 697-70:1, Harkness, J. W. and Vantsts. J. T. (1982) : Virology, In: Uorder disease of sheep: a vlrus induced teratogenic dtsorder. A collaboratorattve review edited by Barlow, RM, and Pat~ tersnn, D.S,P BerUn and Hamburg, Paul Parey (advan<'es in vcterlmuy mcdi Clne.Fortseht1tte der VeteIinarmroizln .. ~)6:56-64 Hassan. H. B.: EL-Sanousi. A.. EL-TrabUl. M. A.: F8thla. M. M.; Ayoub. N, N. K.; EL-Bagomy. G. T. ud Mouua, A. M. (19891 : Serological survey for neutralizing antibody to bovinf' 'vlral diarrhea virus in Egypt. d.t:gypt. VPi.Med,As$" 4BO -2): 3~ 1-3R}t uungerford. T. G. (1990) : DiS('(tses of live &tociU;Uh. F:d,MeGraw-HiII Book Co., Sydney. Huasln. A. A. and Waldehlwet. Z. {1994} : The effect of experimental inl("f'tion ",,1lh boniC'r dis ease virus un lymphocyte subpopulaUon 1n the- peripheral blood ur lambs,R('~'LVd,Sd., 56:20]-207. Lamontagne. L.: Lafortune. P. and Fownet. M. (I989) : Modulation of the <:ellular rcspons('s to T~<:en dependent and T· cell Independent antigens In lambs with indu('cd txwiOf> viral diarrhea virus infectJon Am"J.Vct.Rcs.. 50(9): 1604-1608, Loken; T. (19951 : Border dIsease in sheep,Vct.Clin.North.AnLrood. Anlm.PracL, 11: 579·595, Markham. R. J. F. and RamnaraJre. M. L. (19135) ; Realse or immunosuppresive substum'e from tissue culture cells infceted with bovine viral diarrhea virus.Am.J.VcLRes., 46:879·88:l Mansoura, Vet. Med. J. Vol. IV, No. I, 2002 Hamoda. F. K. et al .... 125 Martin. W. 8. and Aitken. l. D. (2000) ; Diseases of sheep,3rd ED.Blackwell S('ientifk Publica tions Londonu Oxford. Monle&. R. J. and Simpson, V. R. (l997) : Syndrome in sheep resemLliog murosal disease in cattle. Vet. Rec .. 141(171:451 Nettleton. P. F. (lSS8); Border disease update, Vet.Rec.Suppl: In prdcUce., 10: 76. Nettleton. P. F. IWd EntrlcIW. G. (l99S): Review: Ruminant pestiviruses. Hr.Vel.J" 151 :515· 642< Nettleton. P. F.: GUuy. J. A..; Russo. P. and DUaai. E. (1998) ; Border disease or slH'cp and goats. VeL Res.• 29:327-:340" NU.en, A. and Slettbakk. T. (19951 : The connection betwecn the prevalcnr:t' of antibodies to bovine diarrhoea virus/bordcr dism5e virus in sheep and the uovinc diarrhoea virus status of ~attk in combined livestock opcrations. Norsk V!'icnnacrtldsskrirL, 107(7): 765-77L Orr. 114. B. {I9S2] : Puthnlugy of th(' skin and lleece.ln: Border disease of sheep: a vinls-lmiUecd teralogenic disorder, Advanees 10 VeLMed., ::«3: j 3-21. Orr, M. B. and Barlow. R. M. (1978) : Experiments tn border dise OIbum., B. and C&atruecl. G. (1991) : Diaplaccntal Infections with pesIJviruses.ArchVlrol. Supp!.< 3,71,78< Penny. C. D.; Low. J. C.: Nettleton, P. F.; Scott. p, R.; SargIson. N. D.; Strachan. W. D. and Honeyman. P. C. (1996): Coneurn"nt bovine viral diarrhea virus and sahlllmdla ty· phlmurum DTI04 infection tn a group of pregnant dairy hetfers. VcLHec .. 18;485A8~. Plant. J. W.: Acla.nd. n. 14.; Gard. G. p, 8.Dd Walker, R. H. (1983) : Clinical vanatiom. 01 uur der disease tn sheep accordIng to the source' of inoculum.VeLReJ:"., 11~~:58·60. Jliugh. D. G. (2002) : Sheep and Goat med!cine. W.R Saundres Cumpany .. A HarCOllrl tkalth ScierHx's Company, Philade'lphia. US1\, Quinn. P. J.; carter. M. E.: Markey. B. K. and Carter. G. it. (l994) : Cltnie'al vetertnary min'O ulology,Mosby~Year Book Eurupe' Limited, Londun. Ratio8tltS. O. M.; Gay. C. C. blood. D. C. and Hlnchellff. K. w. (2000) : Veterinary medicine A textbook of the diseases of cattle. sheep. pigs, goats and horses, 9 th Ed W.B$aunders Company Ltd, Mansoura. Vet. Med. J. Vol. N. No.1. 2002 Hamada. F. K. et a1. ... 126 Rae. C. A. (1994) : Lymphocytic enteritis and systemic vaseuilHis in sheep. Cand, VrLJ .. 05(lO}: 622-625. Roeder. P. L. (1984) ! Studif's on border disease vil"U$ Infection In sheep.l-'hJ), Tht;Sis. Unlverni· ty of London. Roedel". P. L.; Sweasey. D. and Terlecki. S. (lgg3) : Border disease vlms Inf('cUon of newborn lamb.Or.Vet.J., 139: 126-]36. Rossiter. P. B. and Jessette-, D. M. (1982) : MI(:l'Otltre techniques for the assay of rinderpest vi rus and neutralizaUng antibody. Rf's.Vct.. ScI.. 32: 253~25G. Salem. S. A..: Gabel". A. S. and Abo~EL-Hassan. D. O. (2000l ! Hapid dckctlon of avo and IBn viruses as easuses uf genital infectlon in she('p nnd goats. Rgypt,},A.gn, Res" 78{lJ:D 1- 103. Sallchls. R. and Russo. p, (1990) : hdcdion experim('ntalc de I a~~:nu par un pestlviml"> et 1111(' pasteurella hemolytlca de type T4, Red,Med. VeL, j 66: 143-150. Sawyer, M. M. {1992} ; l3nrdcr disease of sheep: t.he disease In the newborn, adnks(:(:nt nnd adult sheep. Compo lmmunol. Micrblol and iufectious diseases., 15: 17 I 177, Sawyer. M. M. I.U1d Osbutn. B. 1. (19B9) : 1l1lnlUX'lc dysrunction characteristic of border dis case, 11 til lnternatlollal Symposium of the world assoclaUvn 01 veterinary microbluio gists. immunologists and speeiahsts in Infectious diseases, Perugia. Mantm,<1. lIaly: 271, Sawyer. M. M. and Osburn. B. I. {19921 : Long terlll effects of prrsjstcnl pcsi!virw; in/"retioll 011 thyrOld secretions in border dJsease sheep. Proceedings of the second fiymposlum om pesUviruscs: 127 -130. Sawyer. M. M,; Schore. C. E. and Osburn. B. I. (1991) : fionler disease of sheep, Mpccts 101' diagnostic and epidemiolog!cal eunsidcraUon.1\rch, ViroL Suppl., :1: 97- 100. Sharp. M. W. and Rawson. B. C. (1986) : VcLR('c,.119:l28.CHcd by Radmsws ct aL. f2000l Shehata, n.; Balteer. A. M. and Zeidan. S. (1989) : Experimental pathologieal )judy on bovin", viral diarrhea-mucosal dtsease In she.ep. Egypt J.Comp. Palh,Clin. Path .. 2( 11: 34-46. Snowdon. W. A..; ParaonsOIl. I. M. and Droun. M, t. (1975) ! The reaetlon of pn'gnalli (>Wt'S tv moculatlon wHh mucosul dls("use virus of bovine rlgln"J, Compo Paihol., 85:24 1"251. Terpstra. C. (19tH) : Border disease: vlnts perslstence. anUuody response and \ransmi~shm studies. Rt's in Vet.Sci.. 30: 185-t90" Mansoura. Vet. Med. J. Vol_IV. No.1. 2002 Hamoda. F. K. et al. .•. Tbabtl. P.; PronzaroU. L.; nU•• !. E.; GWbC'Jrt. J. II.: Hammaml. S.; Pepin, II. and B:ussa. P. (2002) ! Kxperlmental model of border disease virus injection In lambs: comparatlv(' P3- lhogenleity of pestlvtm5es isolated In l"ralice and Tunisia. Vet. Res .. 3311~: 35·45. Vant$ls~ J, I.: Llnklater. It. A.; Rennle. J. c, and Barlow, R. II. (1979) : g"pC'fililenlal ('hal lenge Infection of ewes following a field outbreak of border dlS("Usc.,U'":omp.PatiluL 85:111-120, Woldehlwet. 1. and. Nettleton. P. F. (1991) : Progeny of sht'cp persistently infected with borner disease vlIus.Arch,Vlrol" 3:267,27L Wolf. O. and Buttner. A. (1994) : Clinical aspects and dlagnoslR of border dI.Sl'ase'l'i.{"razH Prax,. 22:35~38, Wray. c and Roed.er, P. (1987) : Res.Vet.ScL. 42:213. CHed by Penny, C.D.: Low, J.e.: Nellie tor'!. P.F.: Scolt. p,R,; Sargll:mn. N.D.: Stracohan. W,D. and lIoneyman, P,C, (l906): CUIl current Ixwine viral dIarrhea virus and salmonella Iyphlmurum DTI04 lnfrdloll III a group of pregnant dairy he1fers, VeLRcc" 18:485-489. zaghawa. A. (1998) : Prevalence of anUbodles to bovine vIral dlarrhuea virus and/or bOrUer dIs ease virus 1n domestic ruminanls. J.VeLMed,. 45(61: 345<'J51. Mal't$(Jura. Vet• .lied. J. Vol. N. No. J. 2002 Hamoda~ P. K. et 01 .... 128 ~I~I (J.J."JI.;..J") r~"i11J" ~.) ~I""J~ .,$J..wl ~ ,.:A... IJ.I .,;..... rJ ..,.)II1»~! rJL.J1 ,,__ J. rl:.i.':il ~ ~I i~Il.,....J~I..,.lr':il iJ" j,.)"..,JI""r ,-, V'" ul-." r~1 V'" u~ ~I ~.u." w. .~.,..U; z. J II wWIJ ~Uj r~~j ~ ~L,..i ~'jJ ~ · ...... hAIJ ...... A".iJ1 ~I.,-ill ut.,..JI.:;..;..,., "'J 4.hJJ.;JIJ..".,J1 ~ ,JJ-" .) <.i,.i.tl ;.,t.;...J1 ,:,UWj 'il ...... iJ" ",1..0 "L..lI .:.t;I~I ...... "i ...;Jol r l:.i."W ~I ,.,-.ill -UJ ~ J4-!.> l..,..i;i ...,...... vl).,;t 'y.) u...;Joi ut;I~I ...... ;:,IJ .(~}I.iJ.,l5 .;:...... ,l.j~ "r-> ,.,-.ill .,,~' "" ...... ) vl"i}1 iJ"""'; ...:.... J;;.,....;.) """.>lji ~'Y.... Ji 1"'<) ","4.'!' iJ"...,...;.::., .... t.J)l1 ~'};..;.. V"Jr.-iI l 'ftJ "" .s'.;;ll,.; .... J I.) •.).,.,JI) ~I V"J"'; 'ft.);+III.:.t:,..lJ ""}Jr.-iIl J- J rY .:;.;Ll' wl,J1 rl:.i.':il J WI,JI ,,~I J ,,~}I """" ,,';.JI,) ;"';')1 ;.,..;JI "i ","",:"..,JI IJ. ,) 44)1 ~I ut....!....,hAI.,..,.u1 .,)1.,::11.). / ;.... r.)' •• A .;:,l,j}I..,..;.::.:tS L..:" /0 j~ • rAj' yu. rt...,.i 'ftJ....;Jo1 J,WlI r.,.,..J1 j~1 .i.i4N "r-'< ~)WI.) ,,'i.,$ L,.:..,..!.,lIl ",LJH 'YoJ...;Jo1 .... IJ J-r---.) yu. rt...,.! .... "",.; W' "-.t'U>...L,..J1 4.hJL;JI.) "WI rl:.i.'lil r.-') 4.i.Jo;.... ".)Y< i»J?il1.,.4 ;;j.UI .:.!.;,.all .;.. 4}.-JI J .,.}~I ;.,.L...l1 .:.1). ..:> 'il .",...... ,.-JI .;.. ,-, Mansour-a. Vet. Med. J. Vol. IV. NO.1. 2002 Hamoda, F. 8. et al.... 129 ~,,; rL;i.~1 ~.,rJ1 ;#I...,...,~I ","I,..~I V" 0'0..,J1 ",",.. - \ , ,/tl.) J.t....z> .:...,.JI ....~ ,,; y.., # .",UoJI .i.> ,;i .,)j" c,..-:.)I "..,s:,.. .;.."J ".,.....:JI .J...-li ;,.U; "I, ..I·;... ~I ","Ir ', .;.,.. -T ,.:...:.;I.I.J-"",n ,:, Il ,:, )'1-l1 V" ~ J"'<" ,,; "-,,,) I ,;,:1';1... )1 "I, r' WI .J...-li .;:,1; : L;i. '11 ~ ,i ~1 """"";1, ~I ",L...)'I V".;w.,rJ1 ~UI t:' ;;""'81, ~I ~U.I';" ~ 'J'r.J .;.)1 -L ,l.,lVI ,-,10.,)1"; ".,.... ."...i 41 ~I •.lAo, "",..".;J~ ,,"I,,! L,.>"":"'il J!.o :i-,j4--l)'1 vl,J}:.,..ll .....,,; ~,,:,l,I,,.,J.I.i.>"'''''';'' J."!,.,,..;h V".:ui- .u U ) li;':I lor"",.".;J I Ii.> J li;; I ..,; 15:..1 J5 j; ;J,JW 1~ I J-""'" or ".HI J 4-'11 ..,..J"'; J)'- -. ,':/.,,;) W I, ~I, I}.,.. I. d.;":'; """ ."..;:-11 ",o..J1 V" .;L.; .,rJ1 ",l,I"";'1 -, ,),=11 ","A 4::-,L..), ,L;i. ':II J1 1,';U. #1 rl»":""'il-V .iJ..';' vi?! ..... ,..".;JI;,riJ <.+'1 ~I"";l,l i~1 <,$,.ull.oj; ~ <.+':1 4- Ii.>,.,r-,JI "",,,..; or.:.,p ':",-» ..,,15:.."1 .,)j;, w,.,1I:i-,).",J1 vl,.l;JJl ~ • wi r.ro-ll,i ~I t;;l1 J"'<" ,,; re.. ';1""""" ".lJ1 ~I "",.,,; or eli.! ~ -v ,.:;,L.I;JJI .i.> rl~1 ""< ",")1 )L:.;;;I J1 "'j; ·rL;i.~I.,U ...... '0'''';' CT'I rL;i.';1 ~,,; ",I~I, Mansoura. Vet. lIfed. J. Vol. IV. No.1. 2002