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Interaction between chicken anaemia virus and liveNewcastle disease vaccineG.F. De Boer a , D.J. Van Roozelaar a , R.J. Moormann a , S.H.M. Jeurissen a , J.C. Van DenWijngaard a , F. Hilbink a & G. Koch aa DLOCentral Veterinary Institute, Virology Department , P. O. Box 365, 8200, AJ Lelystad,The Netherlandsb Animal Health Service in the Southern Netherlands , P. O. Box 4, 5280, AA Boxtel, TheNetherlandsc Central Animal Health Laboratory , Wallaceville Animal Research Centre , P. O. Box 40063,Upper Hut, New Zealandd Serendip BV, Research in Biotechnology , Jagerseveld 34, 8222, AC Lelystad, TheNetherlandsPublished online: 12 Nov 2007.

To cite this article: G.F. De Boer , D.J. Van Roozelaar , R.J. Moormann , S.H.M. Jeurissen , J.C. Van Den Wijngaard , F. Hilbink& G. Koch (1994) Interaction between chicken anaemia virus and live Newcastle disease vaccine, Avian Pathology, 23:2,263-275, DOI: 10.1080/03079459408418994

To link to this article: http://dx.doi.org/10.1080/03079459408418994

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Avian Pathology (1994) 23, 263-275

Interaction between chicken anaemia virus andlive Newcastle disease vaccine

G. F. DE BOER1,4, D. J. VAN ROOZELAAR1, R. J. MOORMANN1,S. H. M. JEURISSEN1, J. C. VAN DEN WIJNGAARD2, F. HILBINK3

& G. KOCH1,5

1DLO-Central Veterinary Institute, Virology Department, P. O. Box 365, 8200 AJLelystad, 2Animal Health Service in the Southern Netherlands, P. O. Box 4, 5280

AA Boxtel, The Netherlands, and 3Central Animal Health Laboratory, WallacevilleAnimal Research Centre, P. O. Box 40063, Upper Hut, New Zealand

SUMMARYThree groups of 150 SPF chickens were spray-vaccinated with live Newcastle diseaseLa Sota-type vaccine (clone 30) at one day of age, and another three groups wereNDV spray-vaccinated at 10 days of age. In each of the two series of NDV-vacci-nated groups, one group also received at day-old 105 TCID50 of chicken anaemiavirus (CAV) also and another group 105 TCID50 of CAV plus a low dose of virulentMarek's disease virus (MDV). After one week, chickens of the groups which hadbeen NDV-vaccinated and CAV-infected at day-old, with or without MDV, showedsevere respiratory distress, conjunctivitis, drooping wings and ruffled feathers. Aftertwo weeks, wet and inflamed eyes were observed. After three weeks the respiratoryproblems were overcome, but the entire group showed retarded growth as comparedwith the group which had received NDV vaccine only. The 'respiratory sounds' weremilder in the chickens NDV-vaccinated at 10 days of age, about 10% of the chickensshowing retarded growth. Mortality in CAV-infected chickens which had receivedNDV vaccine at day-old was above 30% at 4 weeks of age, and between 15 and 20%when NDV had been administered at the age of 10 days, and was 5% in the twoNDC vaccine control groups. Decreased haematocrit levels were measured in allfour CAV-infected groups at 14 days of age. In serum samples collected for 6 weeksat weekly intervals from chickens of the six groups, no differences were observedbetween HI antibody titres against NDV virus. Thus, dual infection with CAV andlive NDV vaccine did not impair the humoral immune response against attenuatedNewcastle disease vaccine.

INTRODUCTION

In the period 1979 to 1981, several thermostable lentogenic Newcastledisease virus (NDV) strains were isolated in the western Netherlands from broilerflocks suffering from respiratory disease at the age of about 4 weeks (Hilbink,

4Present address: Serendip BV, Research in Biotechnology, Jagerseveld 34, 8222 AC Lelystad, TheNetherlands.

5To whom requests for reprints should be addressed.Received 17 June 1993; Accepted 4 November 1993.

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1987). It was suggested that these respiratory problems might have been causedby thermostable lentogenic NDV strains since such strains were recoveredfrom commercial fowl in other countries (McFerran & Nelson, 1971; Kim& Spradbow, 1978; Yachida et al, 1979; Durham et al, 1980; Meulemanset al, 1981). RNA fingerprinting studies, performed subsequently in our labora-tory, showed that the genomic make-up of these isolates was similar to the livevirus NDV vaccines used in these flocks (unpublished). This observation sug-gested rather a persistence of vaccine virus after vaccination and a possible role ofNDV vaccine virus in the etiology of the respiratory disease. Recently, similarsevere respiratory signs were again noticed in broiler flocks in the province ofNoord-Brabant, and these problems disappeared on one farm from the nextflocks following a NDV spray-vaccination at day 1, followed by a boostervaccination via drinking water at the age of 14 days (JCvdW, unpublished).

Chicken anaemia virus (CAV) may cause in young chickens severeanaemia, subcutaneous and intramuscular haemorrhages, aplasia of the bonemarrow and atrophy of the lymphoid organs (Yuasa et al., 1979, 1983, 1987;von Blow et al., 1983; 1986; McNulty et al., 1989, 1990; McNulty, 1991;Noteborn et al., 1991, 1992). CAV infections under field circumstanceshave usually a subclinical course, but reduce the growth of the chickens(McNulty et al., 1990). The presence of other chicken pathogens, however,may create a different situation. For example, dual infections of veryyoung chickens with CAV and various chickens pathogens such as Marek'sdisease virus (MDV), reticulo-endotheliosis virus, adenovirus, reovirus andinfectious bursal disease virus (IBDV), resulted in aggravation of the mortalityascribed to the other agent (von Blow et al., 1986; Yuasa et dl., 1987; Eng-strm, 1988; Otaki et al, 1988; de Boer et al, 1989, 1992; Rosenberger & Cloud,1989; Jeurissen & de Boer, 1993). We have demonstrated in experimentaldual infections with CAV and different doses of virulent MDV, an aggravatingeffect of CAV on the development of MDV-induced lesions, as shown by in-crease of lymphoproliferative and neoplastic lesions, but only when a low doseof pathogenic MDV was used. The opposite effect was observed with higherdoses of pathogenic MDV (Jeurissen & de Boer, 1993). In experimental studieswith attenuated MDV strains of avian herpesvirus serotype 1, a similar aggravat-ing effect of CAV on the residual pathogenicity of attenuated CVI- 988/Rispensand 988C/R6 vaccine viruses was observed, and this effect was associated withtheir degree of attenuation (de Boer et al, 1992). In order to study the possibilitythat the respiratory problems in the field might have been due to the aggravatingeffect of CAV on the residual pathogenicity of attenuated live virus NDV vaccine('vaccination reaction')} we have performed the studies reported here.

MATERIALS AND METHODS

Chickens

Six groups of White Leghorn strain A (WLA) chickens were hatched from eggs

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AGGRAVATION OF NDV VACCINE REACTION BY CAV 2 6 5

of a production flock of specific pathogen-free (SPF) chickens of the CentralVeterinary Institute, Lelystad, and housed on wooden shavings in six separatecompartments of three conventional chicken houses. Thirty chickens of eachgroup were wing-banded. The chickens were fed with starter and growermash feeds. Drinking water was supplied in stationary fountains. The dailyinspection and maintenance was done after a change of clothes in each chickenhouse, and in the order from single virus-exposed groups to triple virus-exposedgroups.

Experimental designGroup 1, comprising 148 chickens, and groups 3 and 5, comprising 168 chickenseach, were spray-vaccinated with live NDV vaccine Nobilis clone 30 (Intervet,Boxmeer, the Netherlands) at 1-day-old using a Birchmeyer (Flox) apparatuswith 0.55 mm spray nozzles at 4 to 6 bar. One-hundred-and-twenty-fivedoses were dissolved in 500 ml and used per 1,000 chicks. Groups 2, 4,and 6, also comprising 168 chickens each, were NDV spray-vaccinated atthe age of 10 days. All chickens of groups 1, 2, 3 and 4 were intra-muscularly (i.m.) infected at 1-day-old with 106 TCID50 of chicken anaemiavirus (CAV), which had been grown on the MDCC-MSB1 lymphoblastoidT-cell line and was chloroform-treated before use to eliminate infectiousMDV. Groups 1 and 2 received in addition 5,000 cells of a white bloodcell preparation (WBC) (de Boer et al., 1978), prepared from heparinized bloodof MDV strain K (Rispens et al., 1972)-infected chickens. This particular doseof virulent MDV was used because previous experiments had shown thataggravation of MDV lesions by simultaneous infection with CAV occurred onlywith this low dose of pathogenic MDV (de Boer et al., 1992; Jeurissen & de Boer,1993). When the chickens were 14 days old, haematocrit values were determinedin blood samples collected from 12 to 22 wing-banded chickens of each group.The control groups 5 and 6 were spray-vaccinated with live NDV vaccine at 1 or10 days of age, respectively. Control groups receiving live virus NDV vaccineand MDV could not be included because of the number of available chickenhouses.

SerologySerum samples were collected from 20 wing-banded chickens of each group for6 weeks at weekly intervals. Haemagglutination inhibition (HI) antibody titresagainst NDV were determined according to the routine procedures of the AnimalHealth Service in the Southern Netherlands, Boxtel. Eight HI units of NDVstrain LaSota were used.

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266 G. F. DE BOER ETAL.

ImmunopathologyEvery week, six chickens were collected from each group for autopsy andsampling of tissues for immunohistology. Before autopsy the total body weight ofeach chicken was determined. Lung and trachea samples were snap-frozen inliquid nitrogen and examined by immuno-enzymatic staining techniques withhorseradish peroxidase conjugates of monoclonal antibodies HisC7, HisCl, CVI-74.1 and CVI-68.1, which are immunological markers for leucocytes, B-cells,T-cells and macrophages, respectively (Jeurissen et al, 1988, 1989a,b).

RESULTS

Clinical observationsAt the age of 1 week, chickens of groups 1 and 3 which were NDV spray-vacci-nated at day-old and received at the same time CAV, showed ruffled feathers,severe respiratory signs, conjunctivitis and mortality. Several chickens had alsolost their eyesight. At the age of 2 weeks, wet and inflamed eyes were observed,and a number of chickens showed retarded growth. At the age of 3 weeks, theclinical respiratory signs had disappeared. At that time chickens of both groups 1and 3, although the entire groups were homogeneous in size, were smaller thanchickens of control group 5 which had received NDV vaccine only. No differencesin clinical signs were noticed between groups 1 and 3, despite the fact that group1 had also received virulent MDV at day-old.

Respiratory signs were less severe in groups 2 and 4, which had been NDVspray-vaccinated at 10 days of age, and had received CAV at 1 day of age. Therespiratory signs were observed within one week after vaccination as with groups1 and 3. At 3 weeks of age, 'respiratory sounds' could occasionally be heard inboth groups of chickens. At the age of 3 weeks, chickens with ruffled feathers anddrooping wings were observed in both groups, and about 10% showed retardedgrowth as compared to their litter mates and controls. Respiratory signs were notobserved in the NDV vaccine control groups 5 and 6.

Many chickens died during the first 2 weeks of life. Their number wasparticularly high in groups 1 and 3, which had been NDV spray-vaccinated at 1day of age, and had received CAV in addition. The total mortality in these groupswas slightly over 30% after 4 weeks (Table 1), whereas the total mortality ingroups 2 and 4 was, respectively, 20.6 and 15.6% at the age of 4 weeks. The totalmortality in NDV vaccine control groups 5 and 6 was 5% after 4 weeks.

Body weightsThe mean body weights of six chickens, which were collected weekly for autopsyfrom every group, are presented in Table 2. The growth retardation was mostnoticeable in group 1. Haematocrit levels measured at 14 days of age had anormal level of over 28% in only eight of the 65 CAV-inoculated chickens tested(Figure 1), whereas the haematocrit level was over 33% in the control groups 5

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AGGRAVATION OF NDV VACCINE REACTION BY CAV 267

Table 1. Mortality in groups of chickens which were simultaneouslyinfected with live virus vaccine against Newcastle disease, chicken

anaemia virus and Marek's disease virus

GroupNo. Treatment

% Mortality atExposure No. of

at day chickens 2 weeks 4 weeks

1.

2.

3.

4.

5.

6.

NDV vaccineCAVbMDV-KC

NDV vaccineCAVMDV-K

NDV vaccineCAV

NDV vaccineCAV

NDV vaccine

NDV vaccine

111

1011

11

101

1

10

148

168

168

168

168

168

26.4

12.5

23.8

3.6

0

1

32.5

20.6

31.3

15.6

5.6

5.0

"Spray vaccination with live virus NDV vaccine clone 30.b105TCID50 chicken anaemia virus.cMarek's disease virus strain K.

and 6 (Table 2). Unexpectedly, a severe reduction of the haematocrit values wasobserved in only five out of 16 chickens of group 1, the group which was exposedto all three viruses at one day of age.

SerologyFigure 2 shows the HI antibody response against NDV which resulted in a rise inHI antibody titres up to 5-6 log2 within 11 to 14 days after vaccination. Nodifferences were observed between the mean HI titres of groups 1, 3 and 5(Figure 2, panel A) or between groups 2, 4 and 6 (Figure 2, panel B), respect-ively, vaccinated at day-old or at 10 days of age, and regardless of whether thesehad been exposed to NDV vaccine alone or whether they had undergone a dualinfection with NDV vaccine and CAV, or a triple infection, with NDV vaccine,CAV and MDV.

Immunopathology

Immuno-enzymatic staining, using monoclonal antibodies directed against leuco-cytes, B-cells, T-cells or macrophages, of trachea and lung preparations of five

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268 G. F. DE BOER ETAL.

Table 2. Mean total body weights, and haematocrit levels at 14 days of age, ofchickens which were simultaneously infected zvith live virus vaccine against

Newcastle disease, chicken anaemia virus and Marek 's disease virus

Groupno.

1.

2.

3.

4.

5.

6.

Treatment

NDV vaccine*CAV1MDV-Ke

NDV vaccineCAVMDV-K

NDV vaccineCAV

NDV vaccineCAV

NDV vaccine

NDV vaccine

Exposureat day

111

1011

11

101

1

10

Body weight at week

1

56"

57

62

57

57

58

2

58

89

75

102

98

104

3

135

168

156

177

170

171

4

218

233

247

224

243

275

%Haematocritvalue

25.2 ( 7.7)c

17.8 (7.8)

18.1 (7.8)

18.9 (7.8)

35.2 ( 2.7)

33.6 ( 2.3)

"Spray vaccination with live virus NDV vaccine clone 30.bMean body weight of six chickens in grams.cMean haematocrit values at 14 days of age; standard deviation between brackets.d105TCID50 chicken anaemia virus.'Marek's disease virus strain K.

chickens collected at weekly intervals during 6 weeks from groups 1, 3 and 5, didnot reveal differences in staining pattern in degree of inflammation or type of cellsinvolved.

DISCUSSION

In the present study, we describe how an experimental dual infection of attenu-ated live virus NDV vaccine and CAV, or triple infection of NDV vaccine, CAVand virulent MDV, lead to more respiratory problems and higher mortality thanthose observed in the control groups which received NDV vaccine only. Theaggravation of respiratory signs and mortality could not be associated withseverity of microscopic lesions, because the immuno-enzymatic staining of tra-chea and lung sections of chickens collected after autopsy at week 1, 2, 3 and 4from groups 1, 3 and 5, showed no differences in degree of inflammation or typeof cells involved, regardless of whether these chickens had been exposed to NDV

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AGGRAVATION OF NDV VACCINE REACTION BY CAV 269

10 15 20 25 A 30Haematocrit (%)

35 40

group 1 group 2 group 3 group 4 group 5 group G

Figure 1. Distribution of haematocrit values in chickens exposed to Marek's disease virus strainK and chicken anaemia virus at day 1 and spray vaccinated with live virus vaccine againstNewcastle disease at day 1 (group 1, 3 and 5) or at day 10 (group 2, 4, and 6).

vaccine alone or whether they had undergone a dual infection with NDV vaccineand CAV, or a triple infection with NDV vaccine, CAV and MDV. Therefore, nofurther attention was given to the differences in immunopathological lesions, andwe concluded that the aggravation of clinical respiratory signs and mortalityshould be attributed to co-infection with CAV. Since no immunopathological andclinical differences were observed between chickens of groups 1 and 3, we did notobtain evidence for additional aggravation of respiratory signs by CAV in thepresence of MDV. In addition, in several previous MDV vaccination experimentsemploying SPF chickens in the same conventional chicken houses, we have neverobserved the 5% early mortality which occurred in NDV-vaccinated controlgroups 5 and 6.

The aggravating effect of CAV on the pathogenesis of virulent viruses inthe field has been amply demonstrated (von Bulow et al., 1986; Yuasa et al,1987; Engstrm, 1988), and experimentally in dual infections of CAV andIBDV (Rosenberger & Cloud, 1989). We have experimentally demonstratedthe aggravating effect of CAV on Marek's disease pathogenesis as shown by anincrease of lymphoproliferative and neoplastic lesions, particularly when a lowdose of pathogenic MDV was used (Jeurissen & de Boer, 1993). In anothercomparative trial, aggravation and residual pathogenicity was also demonstratedfor attenuated MDV vaccine strains, CVI-988/Rispens and its further attenuatedderivative 988C/R6 (de Boer et al, 1992). In these experiments 7/147 and 1/147chickens showed MD lesions during an observation period often weeks, thereforethe degree of aggravation by co-infection with CAV corresponded with the degree

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270 G. F. DE BOER ET AL.

7-1

g! 5-

i4-= 3-o>

ff

(a)

Jon

titre

inr

.1ra

13en

ffi1

6 -

5--

4 -

3-

2-

1 -

0-

T

0

(b)

^ I0

10

/P/// /

//

i10

i i20 30

20 30Days after NDV vaccination

40

I40

Figure 2. Mean Iog2 NDV haemagglutination inhibition titres in serum samples from twentychickens of groups which were simultaneously infected with chicken anaemia virus and Marek'sdisease virus at day-old, and which were spray-vaccinated with live virus vaccine against Newcastledisease at: (a) day 1 (O group 1; D group 3; A group 5), and (b) day 10 (O group 2; D group4; A group 6).

of attenuation of the MD vaccine strain. We describe here that CAV exerted alsoan aggravating effect on the residual pathogenicity of another attenuated virus,live virus NDV vaccine clone 30.

The aggravating effect of CAV on conditions induced by other pathogenicviruses can be explained by its immunosuppressing effect and/or its enhanching ortransactivating of the virus replication of the other virus.

With respect to immunosuppression we have previously described morpho-logical alterations which might explain immunosuppression. One week afterCAV infection of 1-day-old SPF chickens, a depletion of the thymus cortexwas observed, followed by repopulation with lymphoid cells starting from

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AGGRAVATION OF NDV VACCINE REACTION BY CAV 271

two weeks onwards, and whereas no depletion of the bursa of Fabriciuswas observed Qeurissen et al., 1989a'b; de Boer et al., 1989, 1992; Jeurissen& de Boer, 1993). This effect on the thymus may be the underlying causeof the impaired response to inactivated Newcastle vaccine (Box et al.,1988) and infectious bronchitis vaccine (J. K. Rosenberger, personal com-munication). However, the absence of differences between the mean NDVHI titres in the six test groups demonstrates that the humoral immune response,and thus the T-helper functions associated herewith, against NDV vaccineis not impaired by the combined exposure to NDV vaccine and CAV. Thus,the combined evidence suggested that the aggravating effect of CAV on otherviruses is mainly caused by impairment of other T-cell-mediated immunefunctions.

Various interactions between genes or gene products of human viruses havebeen documented; this has become an important aspect of modern virologicalstudies aimed at AIDS-related problems (Broder, 1992). In the field of avianvirology, Tieber et al. (1990) described differences in transactivating activitybetween high passage and low-passage MDV vaccine strains of serotype 1,presumably caused by differences of activation of endogenous avian leukosisvirus (ALV) proviral LTR promoters, and Pulaski et al. (1992) recently des-cribed the activating effect of MDV serotype 2 on ALV gene expression andvirus replication. By electron microscopic observation of CAV-infected MDV-or ALV-transformed lymphoblastod cell lines, we have observed increasednumbers of MDV and ALV particles, and therewith obtained evidence thatCAV exerts similar activity on MDV or ALV replication in vitro (un-published). The preference of CAV for thymocytes on the one hand andfor haematopoietic precursor cells on the other is of great interest for theunderstanding of the CAV pathogenesis. Taniguchi et al. (1983) have suggestedthat the erythropoietic function of the bone marrow is under controlof T-lymphocytes, so that depletion of T-lymphocytes would cause a reducedproliferation of erythroblasts. On the basis of previous experiments, we sug-gested that competition for infection of T-cells occurs between CAV and MDV.In other words MDV might rescue part of the erythropoiesis due to protectionof an important fraction of the T-lymphocytes from lethal infection by CAVQeurissen et al., 1989a>b; de Boer et al., 1992; Jeurissen & de Boer, 1993). Thedifferences between the haematocrit levels of chickens of groups 1 and 2 arein agreement with such a competition between CAV and MDV for infectingT-cells.

In 1980, at the time of the recovery of thermostable lentogenic NDV strains(Hilbink, 1987), CAV infections were as widespread in the field as these are now(McNulty et al, 1990; McNulty, 1991). In fact, in 1986, one of us (JCvdW)observed CAV signs in SPF chickens which had been inoculated with materialsfrom NDV-vaccinated broilers with respiratory problems. In conclusion, theresults of our experiments make likely that the respiratory conditions in broilersin the field may have been due to the aggravating influence of CAV on theresidual pathogenicity of live NDV vaccines.

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272 G. F. DE BOER ETAL.

ACKNOWLEDGEMENTS

We thank Ms E. M. Janse and Mrs H. M. Boerrigter and J. E. Groenendal forexpert technical assistance and Mrs Victoria Wensvoort-Thatcher for editorialassistance.

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NOTEBORN, M.H.M., DE BOER, G.F., VAN ROOZELAAR, D.J., KARREMAN, C., KRANENBURG, O., VOS, J.G.,JEURISSEN, S.H.M., HOEBEN, R.C., ZANTEMA, A., KOCH, G., VAN ORMONDT, H. & VAN DER EB, A.J. (1991). Characterization of cloned chicken anemia viras DNA that contains all elements for theinfectious replication cycle. Journal of Virology, 65, 3131-3139.

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RESUMEInteraction entre le virus de l'anmie du poulet et la vaccination virusvivant contre la maladie de NewcastleTrois groupes de poulets exempts d'organismes pathognes spcifis (E.O.P.S.) ont tvaccins par nbulisation, un jour d'ge, avec un vaccin virus vivant contre la maladie deNewcastle, de type La Sota (clone 30), et trois autres groupes ont t vaccins de la mmemanire dix jours d'ge. Dans chacune des deux sries de groupes vaccins, l'un d'eux a reu, un jour d'ge, 105TCID50 de virus de l'anmie du poulet (CAV) et un autre groupe105TCID50 de CAV plus une faible dose de virus pathogne de la maladie de Marek (MDV).Aprs une semaine, les poulets appartenant au groupe qui a t vaccin contre la maladie deNewcastle et infect CAV un jour, avec ou sans MDV, ont prsent une forte dtresserespiratoire, de la conjonctivite, des ailes tombantes et des plumes bouriffes. Aprs deuxsemaines, les yeux taient enflamms et humides. Aprs trois semaines, il n'y avait plus desymptme respiratoire mais la totalit des animaux prsentait un retard de croissance compar ceux qui n'avaient reu que le vaccin. Les bruits respiratoires taient moindres chez lespoulets vaccins contre la maladie de Newcastle dix jours d'ge parmi lesquels 10%prsentaient un retard de croissance. La mortalit, chez les poulets infects CAV qui avaientt vaccins un jour, tait suprieure 30% quatre semaines d'ge et entre 15 et 20%

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lorsque le vaccin contre la maladie de Newcastle a t adminstr l'ge de dix jours alorsqu'elle tait de 5% dans les deux groupes de tmoins vaccins contre la maladie de Newcastle.Une diminution des valeurs des hmatocrites a t observe dans les quatre groupes infectsCAV 14 jours d'ge. Les prlvements de srum recueillis hebdomadairement, pendant sixsemaines, chez les poulets appartenant aux six groupes n'ont montr aucune diffrence destitres d'anticorps inhibant l'hmagglutination du virus de la maladie de Newcastle. Enconsquence, une double infection avec le CAV plus un vaccin vivant de la maladie deNewcastle ne modifie pas la rponse humorale conscutive l'administration d'un vaccinattnu de la maladie de Newcastle.

ZUSAMMENFASSUNGWechselwirkung zwischen hhneranmievirus und Newcastle-lebend-vakzineDrei Gruppen von 150 SPF-Kken wurden im Alter von einem Tag und weitere drei Gruppenim Alter von 10 Tagen mit Newcastle-Lebendvakzine vom Typ La Sota (Klon 30) mittelsSpray vakziniert. Jeweils eine Gruppe wurde am ersten Lebenstag auch mit 105 TCID50Hhneranmievirus (CAV) und eine weitere Gruppe ebenfalls mit CAV und zustzlich miteiner niedrigen Dosis von virulentem Virus der Marekschen Krankheit (MDV) infiziert. Nacheiner Woche hatten Kken der am ersten Lebenstag NDV-geimpften und CAV-infiziertenGruppen, mit oder ohne MDV, starke Atemnot, Konjunktivitis, hngende Flgel und ges-trubtes Gefieder. Nach zwei Wochen wurden feuchte und entzndete Augen beobachtet.Nach drei Wochen waren die Atmungsprobleme berwunden, aber die ganze Gruppe war imVergleich zur nur NDV-geimpften Gruppe im Wachstum zurckgeblieben. Doe Atemger-usche waren bei den im Alter von 10 Tagen NDV-geimpften Kken schwcher, und etwa10% der Tiere wiesen eine Wachstumshemmung auf. Die Mortalitt im Alter von 4 Wochenlag bei CAV-infizierten Kken, die am ersten Lebenstag mit NDV vakziniert wurden, ber30% und zwischen 15 und 20%, wenn NDV im Alter von 10 Tagen verabreicht worden war,und sie betrug 5% in den nur NDV-geimpften Kontroll-Gruppen. Verminderte Hma-tokritwerte wurden im Alter von 14 Tagen in allen vier CAV-infizierten Gruppen gemessen.Bei den Serumproben, die 6 Wochen lang in wchentlichen Abstnden von Kken aller sechsGruppen gewonnen wurden, waren keine Unterschiede zwischen den HA-Titern gegen NDVfestzustellen. Die Doppelinfektion mit CAV und NDV-Lebendvakzine beeintrchtigte alsonicht die humorale Immunantwort gegen attenuierte Newcastle-Vakzine.

RESUMENInteraccin entre el virus de la anemia del pollo y la vacuna viva frente alvirus de la enfermedad de NewcastleTres grupos de 150 pollos SPF fueron vacunados mediante pulverizacin con la vacuna tipoLa Sota (clon 30) del virus de la enfermedad de Newcastle el primer dia de edad y otros tresgrupos fueron vacunados mediante pulverizacin con NDV a los 10 das de edad. En cada unade las dos series de los grupos vacunados con NDV, un grupo tambin recibi a la edad de unda 105 TCID50 del virus de la anemia de los pollos (CAV) y otro grupo 105TCID50 de CAVms una dosis baja de virus virulento de la enfermedad de Marek (MDV). Despus de unasemana, los pollos de los grupos que haban recibido vacunas de NDV y CAV a la edad de unda, con o sin MDV, mostraron alteraciones respiratorias intensas, conjuntivitis, caida de lasalas y plumas encrespadas. Tras dos semanas se observaron ojos inflamados y hmedos. A lastres semanas se superaron los problemas respiratorios pero el grupo entero mostr uncrecimiento retardado al compararlo con el grupo que haba recibido nicamente vacuna

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NDV. Los sonidos respiratorios fueron menos acusados en los pollos vacunados con NDV alos 10 dias d edad, mostrando alrededor de un 10% de las aves de este grupo un crecimientoretardado, la mortalidad en los pollos infectados con CAV que habian recibido la vacuna NDVal da de edad fue superior al 30% a las 4 semanas de edad y entre el 15% y 20% cuando dichavacuna se administr a los 10 das de edad, siendo del 5% en los dos grupos control vacunadoscon NDV. Se midieron los niveles reducidos de hematocrito en los cuatro grupos infectadoscon CAV a los 14 das de edad. Se recogieron muestras de suero durante 6 semanas, aintervalos semanales de los pollos procedentes de los 6 grupos no observndose diferencias enlos ttulos de anticuerpos HI frente al virus NDV. En conclusin, la infeccin doble con CAVy vacuna viva NDV no alter la respuesta inmune humoral frente a la vacuna de la enfermedadde Newcastle.

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