DURATION OF IMMUNITY OF LIVE VACCINE CEVAC S.

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1 DURATION OF IMMUNITY OF LIVE VACCINE CEVAC S. Gallinarum TESTED BY Salmonella Enteritidis AND Salmonella Gallinarum CHALLENGES, 11 WEEKS AFTER VACCINATION József Herczeg1, Ágnes Makai1, Imre Héjja1, Éva Balla1, Gábor Szőnyi1, László Makranszki1, László Fodor2, Jérôme Thevenon1, Rick vanoort3 1CEVA-Phylaxia, Budapest, Hungary; 2Szent István University, Faculty of Vet. Sci., Dep. of Microbiology and Infectious Diseases, 3CEVA-Libourne, France

2 EFFICACY OF CEVAC CORYMUNE 4+7 K USED AS BOOSTER VACCINE AGAINST INFECTIOUS CORYZA, Salmonella Enteritidis AND Salmonella Gallinarum József Herczeg1, Ágnes Makai1, Imre Héjja1, Éva Balla1, Gábor Szőnyi1, László Makranszki1, László Fodor2, Jérôme Thevenon1, Rick vanoort3 1CEVA-Phylaxia, Budapest, Hungary; 2Szent István University, Faculty of Vet. Sci., Dep. of Microbiology and Infectious Diseases, 3CEVA-Libourne, France

3 EFFICACY OF CEVAC CORYMUNE 7 K USED AS BOOSTER VACCINE AGAINST NEWCASTLE DISEASE, INFECTIOUS BRONCHITIS AND EGG DROP SYNDROME József Herczeg1, Ágnes Makai1, Imre Héjja1, Éva Balla1, Gábor Szőnyi1, László Makranszki1, László Fodor2, Jérôme Thevenon1, Rick vanoort3 1CEVA-Phylaxia, Budapest, Hungary; 2Szent István University, Faculty of Vet. Sci., Dep. of Microbiology and Infectious Diseases, 3CEVA-Libourne, France

4 LABORATORY EFFICACY TESTING OF SUBCUTANEOUSLY ADMINISTERED CEVAC TRANSMUNE VACCINE IN BROILER CHICKENS József Herczeg1, Miklós Nagy1, László Makranszki1, Éva Balla1, Jean defoucauld1, Jérôme Thevenon1, Branko Alva2 1CEVA-Phylaxia, Budapest, Hungary; 2CEVA-Libourne, France

5 ONSET OF IMMUNITY OF LIVE VACCINE CEVAC S. Gallinarum TESTED BY Salmonella Enteritidis AND Salmonella Gallinarum CHALLENGES, 3 WEEKS AFTER VACCINATION József Herczeg1, Ágnes Makai1, Imre Héjja1, Éva Balla1, Gábor Szőnyi1, László Makranszki1, László Fodor2, Jérôme Thevenon1, Rick vanoort3 1CEVA-Phylaxia, Budapest, Hungary; 2Szent István University, Faculty of Vet. Sci., Dep. of Microbiology and Infectious Diseases, 3CEVA-Libourne, France

6 Salmonella Gallinarum EFFICACY OF CEVAC S. Gallinarum VACCINE BOOSTED WITH CEVAC CORYMUNE 4 K AND CEVAC CORYMUNE 7 K VACCINES, TESTED BY CHALLENGES AT 21 AND 35 WEEKS OF AGE József Herczeg1, Ágnes Makai1, Imre Héjja1, Éva Balla1, Gábor Szőnyi1, László Makranszki1, László Fodor2, Jérôme Thevenon1, Rick vanoort3 1CEVA-Phylaxia, Budapest, Hungary; 2Szent István University, Faculty of Vet. Sci., Dep. of Microbiology and Infectious Diseases, 3CEVA-Libourne, France

10 COMPARISON OF NEWCASTLE DISEASE POWDER VACCINES WITH LIQUID VACCINES IN BROILERS CONSIDERING THE HUMORAL RESPONSE AND VACCINAL REACTION K. Huyge1, et al 1Laboratory of Pharmaceutical Technology, Faculty of Pharmaceutical Sciences, Ghent University, Ghent, Belgium 2Department of Farm Animal Health, Faculty of Veterinary Medicine, Utrecht University, Utrecht, the Netherlands 3Animal Health Service GD, Deventer, the Netherlands

11 The efficiency of spray vaccination methods is severely decreased by: the evaporation of airborne vaccine droplets of the generated aerosols, the wide droplet size distributions, affecting the deposition pattern of the vaccine particles causing adverse direct (respiratory distress and mortality) and indirect (mortality, growth retardation and increased susceptibility to colibacillosis) vaccinal reactions

12 Advantages of dry powder vaccines : inactivation by evaporation does not occur as powder vaccines are not subjected to physical state transitions, production of dry powder aerosols with narrow particle size distributions would enable targeting of specific areas of the respiratory tract (upper respiratory tract for primary vaccinations and upper, and lower respiratory tract for booster vaccinations) thereby avoiding the occurrence of vaccinal reactions.

13 haemagglutination inhibition (HI) antibodies against ND virus induced by powder vaccines did not differ significantly from those obtained with current liquid ND vaccines. powder vaccines were further improved by further narrowing particle size distributions and reducing virus loss during production and storage (unpublished data). Finally, in an in vivo experiment the humoral response and vaccinal reaction of powder vaccines was compared to that of current liquid ND vaccines, which is documented here.

14 Dropret size and Sero-response of SPF-broiler hens exposed to powder and liquid aerosols of live Newcastle disease vaccines at 4 days old (Log2 HI ND titre ± SD) Fine ND powder vaccine: 6.3 ± 0.3µm, 4.4 ± 1.0 Coarse ND powder vaccine: 33.3 ± 0.9µm, 3.4 ± 1.3 Coarse/fine ND powder vaccine (9:1): 37.0 ± 0.5µm, 4.3 ± 1.2 Coarse ND vaccine spray: 10.0 ± 0.25µm, 3.2 ± 1.1 Fine ND vaccine aerosol: ± 20.7µm, 5.2 ± 0.9

15 Discussion Although less virus inactivation occurred in the powder vaccines during dispersion, no significant differences were found between them and the corresponding current liquid vaccines regarding the humoral response. The fine liquid ND vaccine aerosol induced the highest HI titers despite the fact that here the largest virus inactivation occurred due to evaporation, resulting in much lower virus concentrations per m3 air than the fine powder vaccine.

16 COMBINED ADMINISTRATION OF NEWCASTLE DISEASE AND AVIAN METAPNEUMOVIRUS LIVE VACCINES TO ONE-DAY- OLD TURKEYS: VACCINE INTERACTION AND PROTECTION AGAINST VIRULENT AMPV Caterina Lupini1*, et al 1Dipartimento di Scienze Mediche Veterinarie, Alma Mater Studiorum University of Bologna, Ozzano Emilia (BO), Italy; 2Dipartimento di Sanita Pubblica, Patologia Comparata e Igiene Veterinaria, University of Padua, Legnaro (PD), Italy; 3Istituto Zooprofilattico Sperimentale delle Venezie, Legnaro (PD), Italy p685

17 Inroduction Vaccination against Newcastle disease (ND) is compulsory in Italy (6) and turkeys are also frequently vaccinated against Turkey Rinotracheitis (TRT, Avian Metapneumovirus (AMPV). Live vaccines administered during the first days of life with at least an interval of 7 days between the vaccinations, so as to avoid possible interference between the vaccines. A vaccination programme including a simultaneous administration of both vaccines in the hatchery would have several practical, economic and biosecurity advantages. Such a modification requires a study about the vaccines compatibility.

18 Treatment groups TRT group (16 birds) vaccinated with strain AMPV subtype B VCO3; B1 group (8 birds) vaccinated with strain NDV B1; VG/GA group (8 birds) vaccinated with strain NDV VG/GA; TRT+B1 group (16 birds) vaccinated with strains AMPV subtype B VCO3 and NDV B1, coadministered; TRT+VG/GA group (16 birds) vaccinated with strains AMPV subtype B VCO3 and NDV VG/GA, co-administered; CONTROL group (16 birds) treated with sterile water.

19 Sampling Oropharyngeal swabs were collected at 2, 4, 6, 8, 10, 14, 18, 22, 26 and 30 days p.v. to investigate the colonization and replication of vaccine viruses in target tissues by reverse transcriptase (RT-PCR). Blood samples were obtained at 7, 14, 21 and 28 days p.v. for detection of antibodies anti-ampv by IDEXX ELISA,

20 Challenge At day 21 p.v. eight animals from groups TRT, TRT+B1, TRT+VG/GA and Control were moved to other isolation units and challenged with virulent AMPV subtype B strain IT/Ty/Vr240/87. Each bird received by eyedrop 3.77 log10 CD50. From day 1 post-infection (p.i.) until the end of the trail, birds were monitored daily for clinical signs assigning a score to each bird, following the method described by Naylor and Jones (5).

21 RT-PCR results on vaccine take Number of positive birds/day of sampling to the RT- PCR for NDV were comparable in all vaccinated groups. All the animals shed at least once 4 days p.v. and the number of positive birds /group decreases significantly after 10 days p.v. The total number of positive birds to RT-PCR for AMPV differs only slightly among the three vaccinated groups. All animals shed at least once on 14th day p.v.

22 Serology No statistically significant difference (p> 0.05) between antibody titers of the AMPV vaccinated group and the control group until day 21 post-vaccination was found. At 28 days post-vaccination the mean antibody titers appears to be statistically higher in groups vaccinated in combination when compared to the Control group and the TRT group. The mean NDV-HI titers did not show significant differences between groups until day 21 (p> 0,05) (Figure 3). At day 28 the control group and the vaccinated groups differed significantly (p<0,05). Not statistically differences were observed between groups that received single or in-combination vaccines for bothvaccinal trains.

23 Challenge results After challenge with field AMPV strain, unvaccinated and challenged birds showed clinical signs with a mean score of 15.25, significantly higher than in all vaccinated groups. No statistically significant difference in clinical scores was found between the groups vaccinated in single or in combination.

24 In conclusion, It appears that concurrent vaccination of one-day old commercial turkeys with AMPV VCO3 vaccine strain and NDV B1 or VG/GA vaccine strains gives the same protection after AMPV challenge. Furthermore the AMPV vaccine given alone do not interfere with vaccines replication in the respiratory tract.

25 EFFECT OF CHALLENGE WITH 3 VIRULENT NEWCASTLE DISEASE VIRUSES ON EGG PRODUCTION IN VACCINATED WHITE LEGHORN LAYING HENS. Ruben Merino1, et al Departamento de Medicina y Zootecnia de Aves, FMVZ, UNAM. Mexico, DF, 04510, Mexico Merial Mexico SA de CV, El Marques, Queretaro, 76246, Mexico p701

26 Introduction Outbreaks of ND in birds older than 4 weeks old can be seen in two forms: respiratory distress is common in well protected flocks, with low mortality and digestive or nervous symptoms (1-2%), the diagnosis can be difficult. The classic form of ND is seen in flock with non-uniform immunity, so, nervous and digestive signs are common as well as drop in egg production and mortality rate higher than 5%

27 Vaccination program Age (w) Groups A B C vaccine 1 IB / ND ConnMass/La Sota strain ConnMass/La Sota ConnMass/La Sota 3 ND LaSota LaSota LaSota 5 ND+IB Coryza 127 5HpABC LaSota/Mass HpABC LaSota/Mass41 5 ND LaSota LaSota LaSota 46 IB - ND ConnMass/La Sota ConnMass/La Sota 10 ND P2005 P2005 Ulster 127 5HpABC LaSota/Mass41 ConnMass/La Sota 15 ND+IB+EDS Coryza 127 5HpABC LaSota/Mass HpABC LaSota/Mass41 15 ND P2005 Ulster 15 ND LaSota LaSota

28 Challenges 25 hens from groups A, B and C where challenged with Chimalhuacan, Torreon or Mx14 viruses, at 25 weeks old, by the ocular route with 106 EID50% / 0.2 ml After challenge, all groups were observed for 4 weeks. Samples for virus isolation were taken from 3 hens at 2 and 4 weeks post challenge. Virulent standard challenge strain Chimalhuacan, as well as Torreon strain (responsible for the major outbreak occurred in Northern Mexico in 2000) and a contemporary strain isolated in 2008 (Mx14 strain) where used for challenge.

29 Challenge results Egg production was above 90% at challenge, for all groups. The mean egg production in the 4 weeks after challenge period was lower in A group than in B and C groups. Egg quality was better (P<0.05) in groups B and C when compared with group A, regardless of challenge strain. Mortality was higher in group A than in the others, but only statistically significant in the challenge with Chimalhuacan and Mx14 NDv strains. All hens from group A showed clinical sings, and they were more severe than those observed in the other groups, independent of the challenge strain.

30 Serology The geometric mean of the titer in group A (HI titre at 24w: 38) was significantly lower (P<0.05) than in groups B and C (HI titre at 25W: 1702 and 2426), at the laying hens reception and previous to challenge. After challenge, there was not statistical difference among groups, independent of the challenge strain.

31 EFFICACY OF SEVERAL VACCINATION PROGRAMSin day-old pullets (Lohmann breed) AGAINST NEWCASTLE DISEASE CHALLENGE. Satra, J.1, et al 1Veterinary Biologics Assay Division, Bureau of Quality Control of Livestock Products, Department of Livestock Development Pakchong, Nakhonratchasima Thailand 2Ceva Animal Health Asia Pacific, p909

32 Introduction The association of live and inactivated ND vaccines in ayd-old-chicks has been extensively investigated and the results show that the HI titers, protection against challenge and persistence of immunity are better achieved when the combination of live and inactivated vaccines is used, compared to live or inactivated alone The benefit of the combination of live and killed vaccine in the hatchery is particularly clear in a context of strong viral pressure as it strengthens and prolongs the protection by combining the local immunity provided by live attenuated vaccine with humoral immunity (circulating antibodies) conferred by inactivated vaccines.

33 Treatment groups Group A: Vector HVT-NDV vaccine by SC injection (0.2 ml per chick) Group B: Live apathogenic enteric strain ND vaccine by eye drop (ED) route (0.03 ml per chick); Group C: Live apathogenic enteric strain (ED) + Vector HVT-NDV vaccine (SC); Group D: Live apathogenic enteric strain (ED) + Inactivated ND vaccine3 (SC, 0.1 ml per chick). Group E: Positive Control (unvaccinated but challenged with virulent NDV) Group F: Negative Control (un-vaccinatedand unchallenged)

34 Challenge The reference Thai NDV challenge strain (Lopburi strain ICPI 1.86) was used as a challenge virus for the groups A, B, C, D and E. The challenge was carried out by intramuscular application at a dosage of 5log10 EID50 per chick. Day-old chicks from group F were not challenged to serve as unvaccinated and unchallenged controls (negative controls). The chicks were kept in isolators, observed during 14 days and clinical signs and mortality recorded.

35 Serology samplings At day of age, before the beginning of the trial, 20 surplus birds were bled and the level of the maternally derived antibodies was assessed by HI test and a commercial Elisa test kit. At 14, 21 and 28 days post-vaccination, 20 birds per group were sampled and the sera were also tested using both HI and Elisa test. Finally, 14 days after each challenge, blood samples were taken from each of the survivors in all groups.

36 Challenge results Birds challenged at 14 days post-vaccination: All groups showed a very good level of protection varying between 90-95% and without any major differences among them. This is probably due to a high residual amount of maternal immunity.

37 Challenge results Birds challenged at 21 days post-vaccination: At three weeks after vaccination, some differences among the vaccination programs start to be noticed. The unvaccinated control group still showed 70% and this result was very similar to the one obtained by the group B, which was vaccinated only with a live apathogenic enteric strain. The groups A, C and D showed a very high level of protection, particularly the group A which achieved 100% of protection.

38 Challenge results Birds challenged at 28 days post-vaccination: At four weeks post-vaccination, the protection rate of the unvaccinated group was only 10%. The group vaccinated with only the live apathogenic enteric strain (group B) achieved 55% while groups A, C and D had very high levels of protection. Only the groups vaccinated with vector HVT-NDV reached 100% of protection Serology: look at the abstract

39 EFFICACY OF A RECOMBINANT NEWCASTLE DISEASE VACCINE (VECTORMUNE HVT NDV) BASED ON CLINICAL PROTECTION AND SHEDDING OF CHALLENGE VIRUS IN BROILER CHICKENS V. Palya1*, I. Kiss1, T. Tatar-Kis1, T. Mato1, B. Felfoldi1, Y. Gardin2 1Ceva-Phylaxia Co. Ltd., Szallas u. 5., 1107 Budapest, Hungary 2CEVA, Libourne, France p757

Vectormune ND a step towards control of Newcastle Disease

Poultry Vaccinology Summit April 28-30, Budapest, Hungary Vectormune ND a step towards control of Newcastle Disease Yannick Gardin 1, Vilmos Palya 1, John Elattrache 1, Marcelo Paniago 2, Christophe Cazaban