AVIAN DISEASES 42.173-178, 1998 Research Note— Efficacy of Combined Killed-In-Oil Emulsion and Live Newcastle Disease Vaccines in Chickens Raphael Folitse, D. A. Halvorson, and V. Sivanandan Univenigy of Minera, Collegeof Vercinary Medicines Depactinene oF Veterinary Hath Blegy, 1971 Commonmalch Avene, 'e Paul, MN 35108 Received 25 Febnaey 1997 SUMMARY. Following he inode’ route vation miner Wi die types of Newarde dea (4D) vane, de inience of velogenc vaceravopicNewanle {ites (VVNI) i connec poultry wonduige hoe dechasd deme, Unioronersy, ‘he vaesinion tegioes ate a1 feeble in eae wnlDachyan stems of poly prodoanon prctced in many loping nse ii tly re oh tly Cog ssa pm Ale ND me «» do lomacing high el of ND vse (NDV) snes sdegute mont ice Spine ND. Thc led of tail sopomey at mentced by dhe henapfuhaoibion iy ta, avd the dager of prouction apsout the vident sos of NIV ere sided in ‘Biche imal wid difscne acs The wasines aod we: level esaion {ethene cc) ps Ive ru (occa on), gen concur: expermeelwcene le an gop nanny dio Ge vet wan par Gy negra erin rset ee clones edt Gay "he ve ened i hx dy indict thet cncuenesdneon oo smslion sed fee NDW yeti indeed te antsy mega oe ts wa no gions keene n pose anong the vaca owen RESUMEN, Hous de Ineeigsci Eficacia de la combinaign de ons vacana inacivda ‘rmulsond eh ache y una vacuna va crews enfermedad de Newcae en pales Deipas de la ntroduccin de programas ratinanos de vacanaaign con dicents pos de scant cont Neweare, [incidence de fy enfermedad de Newcastle welogeneo vscero- tropes (VVND) he deciondo dramaticancace eth wicltura comercial a nel sana. Denfortinaamonee eioe piogiss ce vacinion ha sm powible on semas de ps0™ ‘duccbn sical de gato y de campo abst practcador en michoe pte en dese Fn cse carci se buses destrollr un stem de vcunacién simple capa Nevexde para ‘rps on trl aley chamderg de dnrinunpos toot vid do Kcaftomedad de New Basle. sdecuads pars protege: Mes pallos conta Ia eafeunead. Se cnudio ol nivel de hy Fapatea de antcuepor mudidoe mediante la prusba de thibicion de le hemaglrinods (ED yd grade proweccion comes cepas siralenes del vias de Neves en pollo in ‘anizadoe con dftentewacuna, Lae vaeum ulead Reron wicuna naetirada ene fas en ace aplcals vis subecines, nds wna vaca iva apliads vs Gelonal ada pisndse al muro temper vacuna exparmentl apieada tn cabaatinaa mar una vac 3 ras viv apliada vis eulooaal y adavisistraln a msn leapo, vacuna eaperatata Hnoctheds Gnulnonsda x aeute pospera svediante la Romogenzaclen de vacua ws ‘comercial y enulsiooads en aceite y ana vaca viva Lo raultadesobxenids cn ene endo fndisan qué i admntractan salen de vacunasinaccdae y exulionadas eh ade y ‘acute viva inducen una rexpuesa mayor de atncaczpor, sin cans, no ube una d= Tiree ignite sn peseeatin ae oe disentergraposvaewnaoe Ky wosee fee-ange chickens, Newstte disse, vaccination SConespondiog aut 13174 Abixeriaions: ELD. ~ eobyo lethal dose 50%, GNET = geomeuic mean Ges ‘urmagghsastien snkibiten; ND ~ Neweaale dasa: NOV ‘eulonaal: VND hemagglutination, HL Newest disease virus, a Tn many developing coneuries, small-sale seating of chickens constitutes a significant pro portion of the poultry industry, even thoragh 3 commercial poultry industey exes. in exe vile ages of these countries, mort Families ease sarall numbers of chickens, carly execeding: 50 birds per household. In this system, a kind of housing is provided in which the birde sleep at fight. Thete chickens seavenge for their feed bar their diets are aFien supplemented wich c ble waste. The bread of chickens kept is usually indigenous burt is sometimes improved with move meaty imporied breeds ‘Backyard pouleey provides an imporcane source of income acd highly animal pro tein wich Title or no capial investment (1, capecally for the rural population. Hewever, this systems is not very efficient Because there ie Tile or no diseate contra. ‘Although the indigenous breeds have a rep seacion for hantiness and resitance «0 divers (9,31) the curvival of these locks remains un’ certain. due to high meriality, which is well aver 5006 per year (9.30). In Gans, velogenic vise cerotropic Newcastle dssase (VND) ad in ‘egnal parasites appear to be the major causes of mortality, Outbreske of VND in backyard ceke remain a potendial threat 19 the eom- mercial pouluy todusuy. ‘Vaccination against Neweasle disease (ND) Jes common practice worldwide and has been ‘asad succesfully for ie prevention and eoneol fora long time (0,12,16) Although vaecination suategies have been avalabie for many years fad have schieved a aignifant success in ro “huetion of VUND owtbreaks and mortals in commercial pouley (8), there has been ack. Df appreciable success in vaccination of fee Fange chickens inv many developing, countries, This may be duc co small oumbexs of chickens per household a lack of understanding, of vax ination among the rural population, poverty, SiPiculey of access to some villages by veer faty personne), improper siorage of the vac tines heavy and carly exposure to infection by bighly virulent Newesile disease virus (SDV) Velogeni visesoupic Neweasl dis strain, andl anexplained vagaries of VWND ope idemiology in the fick, ‘in Ghana and many other African countries kelled-incoil exmulsion ND vaccine ie preenthy the mot widely used vaccine for the prevention dnd conttol of ND in Iree-sange and backyard poultrr Vaccination ie often done twice per year If chickens could be fully protected for a Jong duration by a single ancl vaccination, might significantly rechice ND ubreake and tminimine the cout of vaccination, This might be pomible by concurrent administration of ire sctivated and live virus vaccines, Ie has been reported tnt a comisination of inactivated and Tize virus vaccines has been use 10 siccesfilly Jmnmunize aviary birds (15,19,29). has also been reported tet eoncurtenc application of in- sctiate oil emulsion and live ND vaceine #9 T-day-old broilers has been studied intensively sand proved eo induce snffcioaeimmunicy thet is susealnad for che fattening period (3,202) (Our sihimate goal is the development of a reglive sulsble for single annual vaccination fof ticerange and backyard chickons t9 obztin the desied iminunigy and protection agaane ND. Therefore, the objective of this seul was co develop a miitable onctep vaccination strat fg thar will improve the responce to Killed i fl vaccine alone, The kypathesie tested in chie study was dhat the efficacy of ell-emalion ND. vaccines eatt be itoproved by concurrent ad= tinictracion of live NDV vaccines MATERIALS AND METHODS Chickens. Onedsy-old whive leghorn chicks ner sbuined and rated io a disease bung sien the appropnate fovding and ashondry They snc 4 wheat che su of the capriment. Chik She umroinid wach imzeoned vacones and the Srcrl grap mined in de sme baling al Spo epe in acta copes ated for the Ree Tha namie with lve recta were bused ina di foro balding and lap in negatives lai tots, Bach group was kept in diferane volo, Fou sree ar nctnation ll chickens weve trance Tain Hola oldip sid lege gerbe in 9aSlagle NOV wasnt eine in chickens fem on concn oor. Drinking wnt and com terial ed wee proved ad fn Conmerdal ND vaccine, The ected vas Fas res Valo cmon vaccine coma {sky maoctctred hy Vedand Laberatnes (Vine lind NI} The he vaccine wes Gomera ms Gewied fos NDV- pe Bl Lt Sou sein by obey nit Helle. endora Fgh MNS. Bis ve cle wa erasure jut ptt ce Txperlmemial vaccine. One vil containing 1etd dee of th oped Ie aceme eationed dle ms coi fn 100 al dione wate he vlune of the recostiuad vaccine wa he Fogel with ooe volume of ol cousin ising Sty stnpencs wih por coves Baer Hel fee Cory, Nolen, CA). Te ll aan was 3 bute of eget vom of Moatinde 198 390 ad Mevwside A206 (ach wannfactnel by Sp Tet Fane) Te evn nnn poe fe pees and he vaccine we pepe fp Challe virw NDV GD Tens sin ith 2 Mies conaneron of 10" eyo ia de 3086 ELD.) per bid ns sed for cllenge sy xperiuenal denn, Av of ey the chick eer nda seed no rou banded fe Veco aed bd far proton tre Ficenehclen wane asigaed te cach grup, re ‘lial chichens wre sided" echo he ve Seee props w cna ck Thee yee 32 teament rng shown tgp scene naccouy fe) with de led culm me Gn 7 grap 9, ted ce wih Elena soa fen vacrne pus siouhancos ve vino gen Init; os 3) grag eed ce aah ops fRewal secine 4) gioup fy ewe se wih Gpeinensl vrioe pls simular te vacone a 5) group 5 xed oa wth evn sad rep 6th umacensvl ctl anima Tlic ocine ox ghet at dase fOr geri experimen vate a0 al pe bid and Froid ive vis O4 tl per Dd. The ae ae cong pet bd wi eae tn cost one Elbe of ach vaccine azording tothe managers acs wm not pried: howe dhe United Sates Departmen of Agiite (USDA) eam dose FMDV Ine vactncss 10° HDy (7-209 pri Te tandnd ove Fortes! NDV vaccines ID" ELD, per ad 17) ‘hallo sty Aik fet acini il of Be til wee cnc wo the baalin belay. nd il birt var chlenged wath the velop Sear tuple NDV Tess a ata ce of 10 ELD, nraruacdaty (um), Protection as det tube fora pand of 4 digs air clogs The fhraneter und co era protston fet NDV he ids showing nwo ign and doa bi 175 This ra recorded as mecbidity and woreins 1 sectivey The protection index war seulted se te erent mosbidty and metal in the conta ro Iinus that inthe ceperimenal group, dvd by ee Potent protien inthe conto. gioup, mulled by 100, Serology. Blood saps fo enon ofthe = rum hema unstion-inbbicon (HD) analog level sguinee NDV nese taken prior eo wacinatiny 422 skater vaccination, a wi after vaccine fie {6 challenge), and oe 2-vk alte calles. An Hts ‘ras performed on ale up by tsi 10 het Agsltinason (HA) units of NOV angen ad two {Ul serial dilasons of sera eartig work 10. Goo spate mein citer (GMT) weieeaealate for exch a ‘Statistical analysis, The lol of antibody ee sponte was amlyaed and compared beren gicupy bing + swo-indcpandearsanple wes. Motbidy tnd morealty were anayed by compaction of chi square values, Diferences beeen Os Wee ‘Slered ignicne st = 0.03. RESULIS Anuiboxly response of chickens to veecinasion swith ND yeccines and the resistance 10 chal- lenge with NDV GB, Teens stain, are shown in Table 1 Te was abserved chat the HL GMT value at ME days alice vaccination were high in all of the vaccinared groups exzope that treated sith the experimental vaccine. To the later ea, the re corded value of the GMT was les than 10 ‘Twenty-eight days after vaccination, the H) OMT valucs for the groups vaccinated vith combined killd-in-oil arulsion plus live vitur vaccines and chese vaccinated with killed-inoit emulsion vaccine alone increased rapaly: Th ‘comparison vith the HI GMT valacs obtained 2c 14 days following vaccination, the sncveases ‘were statistically sghifcane (P = 0.05) in both groups, The unvaccinated conuol group had no ddeceetble NDV antibodies. The best antibody respoate war induced by the combined killed ioil plus live vine vaccines After challenge, ic as observed that all of the vaccinated birds were adequately proreted All vaccinated groups recorded 100% protec ion, ith the exception of the geoup vaccinated (3.0) with the experimental vaccine alone, from which two sick birds were eecards, Unvaccin- ated bieds were highly sazospuible to challenges 12 sick and dead biede were recorded,176 Table Noweasle dese [ND) vaccines HGMP Groups 1 dat Bdpm 7 Ty 104 259 ea uw Bs 108: Bu ie “ ° i emulsion voce Fol Geomette mean ts: (GM) and protedon again challenge in chickens vasinted with % Mochity Protection Tedpe® end monty ina 7 © 100 220 ® 109 an 3 4 3 i 100 e © 100 320 a read nibestancouly (sc) with Ella emulvon vied: group 2 = ewe se wie = pts lve cazcine acilopwrally (or), gen aooueanecudy prep 3 ~ ewes ssh experimental vecine)goup 4 ated sich eapeimental racine pls ve viene (OD. Se Simultaneous: group 3 = tated with Iie vie fo) rete: snd group ‘umvacinsted cota AHL = hornggluintioninhbiion, The yas sven is he weiproal ofthe HI GAMT bse on 13 chickens er group, “apar = dave pontacsinson. Sipe. = dvs posicllenge "us inthe same clue ollowed by diferent lowvias suprsros Kes ae siguiianly difeene (P < 005, DISCUSSION Although ic has been established by many researchers that inactivated il emulsion a fines used in the prevention and contiol of poultry diseases can elicit high levels of cise lating 2ncibodies and provide adequace protec tion of bisds agains challenge (6,10,22,23.25), various aterapts ave beew made in roe years to improve the ficacies of such vaccines (723.24). The resules obtained in. this stady indicare th dhe antibody response induced by the com- bined kiled-in-ol vaccines admingeered con currendy wih fue vinoe wae better than thae induced in all other groups, ‘These liferences ‘ere staistcally significant (P< 0.05), Thc ex perimental vaccine administered concurrently with live virus induce? 2 berer aasibody re- sponse than the experimental vacsine adminise tered alone, The difference was sentisicall sig nificant (P < 0.05). ‘We observed thatthe protetion induced by the experimeotal vaccine adminiscered concur reacly with live vurue we beste than thac in- duced by the experimental vaccine adminisened alone, This further conficmed che achantage of the combined vaccines over che oll emulsion Some easier researchers reporeed chat in comparison with live ND vaccines inactivated fil emulsion vaccines indusad long-licing, high, and uniform levels of cireulating an ibod ies in birds shat hal been prapery primed with live vaccines (2.4,18,21,25.29) Inthe presen sindy, we speculate thae the reason for the high antibody eesponte obtained with lulled in-o Ge) plas Ihe vies (on) vaccines give se rmulatancously could be har initially, the Ire vieus replicates quickly in the mucosal mea brane af dhe conjunctiva and che nostily ele iting 2 primary immune reeponse, This i fo) lomed by 2 continuous slow telease of the lille virus ancigen trapped in the oil medium, dhis allowing the hiled virus antigen to behave aa booster dose. This was evident when we com pared the response sr 14 days after vaccination [GMT = 104.389) co thar at 28 days ater ve nation (GMT ~ 258.50. (One disadvantage of mactivated oil emulsion fover live ND szceine: is that inaexvared ol emulsion vaccines donot induce Ines! inom ity in the respiratory and digestive erat, and miu is exablishod rather slowly (29). This Aisedvantage may be minimized by che concur rent administration of lalled-in-oil emulsion and live ND vaccines ‘The experimental vaccine was the least effce tive ofall weatments in antibody response and protection. Many fetors could accou fortis, For example, chore could have been 4 Jow an Ligen dose in the inoculum. Another possibile is that the virus particles might have been n=Siugle NDV vaccination regime in hing sctivaced inthe ol ermulsion and therefore, did not replicace im the dssacs. This latter possi - ity was confiemed by the fact thet contact bites sroconverted Iehas been reported by some reseachers chat tue of inactivated NDV vaccines will provide mo of immunity in pouley (19,29), and that inoculation wich 2 Bee virae sacking fol- lowed in 2-3 wh by an inactivated vaccine imighe provide 9-12 me of protection (11.19) cha been established that primary vaccination, wth insctivacd oil emulsion NDV vaccine of layers 2€ 6 w of age is practical and induces a Sttong immunity thae holds for che entire cow posi life span (23). Trcause 2 higher level af antibody response was obtained in dhe prcsent study with the combined billed in-oil (Se.) plus. live vitus (on) vaccines given concurrently, wwe could speculate chat the ducatien of immunity worl be extended. 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