Director Biology Innovation Strategy Department. Ceva Sante Animale, France
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1 Dr.Yannick Gardin Director Biology Innovation Strategy Department. Ceva Sante Animale, France Graduated as a Doctor in Veterinary Medicine, from Veterinary School of Lyon France 1977 : Join Ceva as Technical Director and Later became in charge of the Marketing and Business development in Latin America 2005 : In charge of managing gthe offer of poultry vaccine He has a strong network of relations within the poultry industry and is a member of the Poultry Veterinary Study Group of the European Union (PVSG EU.) Topic : New Solution for Controlling ND and AI in Asia
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3 Hatchery Vaccination Summit Asia Jeju Island, South Korea, April 26, 2012 NEW SOLUTIONS FOR CONTROLLING NEWCASTLE DISEASE & AVIAN INFLUENZA IN ASIA Dr. Yannick Gardin, DVM Director Innovation Strategy Department, Ceva Santé Animale, Libourne, France YG
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8 NEWCASTLE DISEASE & AVIAN INFLUENZA ARE THE MOST DEADLY DISEASES IN POULTRY 06-00
9 THE 2 BAD BOYS OF A NASTY FAMILY! 07-00
10 ND & AI : 2 Challenging Diseases and an interesting history 1926 : Newcastle upon Tyne, UK (Doyle) Java, Indonesia (Kraneveld) Mid 30s : USA 08-00
11 ND & AI : 2 Challenging Diseases and an interesting history 1926 : Newcastle upon Tyne, UK (Doyle) Java, Indonesia (Kraneveld) Mid 30s : USA Haemorragic syndrom (pseudo) Pest / Plague <-> Avian Influenza) 09-00
12 ND & AI : 2 Challenging Diseases and an interesting history 1926 : Newcastle upon Tyne, UK (Doyle) Java, Indonesia (Kraneveld) Mid 30s : USA Respiratory problems (<-> Bronchitis) Haemorragic syndrom (pseudo) Pest / Plague <-> Avian Influenza) 10-00
13 ND & AI : 2 Challenging Diseases and an interesting history End of the 30 s : 11-00
14 ND & AI : 2 Challenging Diseases and an interesting history End of the 30 s : NDV & AIV are 2 different diseases / 2 different viruses 12-00
15 ND & AI : 2 Challenging Diseases and an interesting history End of the 30 s : NDV & AIV are 2 different diseases / 2 different viruses but with some relationships and similarities il iti 13-00
16 ND & AI : 2 Challenging Diseases NDV AIV 14-00
17 ND & AI : 2 Challenging Diseases MYXOVIRUS NDV AIV 15-00
18 ND & AI : 2 Challenging Diseases MYXOVIRUS Paramyxovirus Orthomyxovirus NDV AIV 16-00
19 ND & AI : 2 Challenging Diseases MYXOVIRUS Paramyxovirus Orthomyxovirus NDV AIV - Rubulavirus (Mumps p virus, Para Influenza) ) 17-00
20 ND & AI : 2 Challenging Diseases MYXOVIRUS Paramyxovirus Orthomyxovirus NDV AIV - Rubulavirus (Mumps p virus, Para Influenza) ) - Morbillivirus (Measles, Distemper, Rinderpest) - Pneumovirus (Respiratory Syncitial V.) - Matapneumovirus (Rhinotracheitis) 18-00
21 ND & AI : 2 Challenging Diseases MYXOVIRUS Paramyxovirus Orthomyxovirus NDV AIV Influenza Viruses
22 Portraits of the 2 criminals NDV μ μ
23 Portraits of the 2 criminals NDV AIV μ μ
24 Portraits of the 2 criminals NDV AIV ENVELOPPE FUSION F HAEMAGGLUTININ HA NEURAMINIDASE N MATRIX HAEMAGGLUTININ NEURAMINIDASE HN RNA 22-00
25 The criminals at work NDV & AIV 23-00
26 The criminals at work NDV & AIV Attachment 24-00
27 The criminals at work NDV & AIV Attachment Penetration 25-00
28 The criminals at work NDV & AIV Attachment Penetration Synthesis 26-00
29 The criminals at work NDV & AIV Attachment Penetration Synthesis Assembling (In the cytoplasm) 27-00
30 The criminals at work NDV & AIV Attachment Penetration Synthesis Assembling Release (by budding) 28-00
31 THE KEY FACTORS OF PROTECTION 29-00
32 The criminals at work NDV & AIV Attachment Penetration Synthesis Assembling Release (by budding) 31-00
33 The criminals at work NDV & AIV ENVELOPPE Attachment Penetration RNA 33-00
34 The criminals at work NDV & AIV ENVELOPPE FUSION F Attachment Penetration /NDV HAEMAGGLUTININ NEURAMINIDASE HN / NDV RNA 34-00
35 The criminals at work NDV & AIV ENVELOPPE Attachment Penetration FUSION F /NDV HAEMAGGLUTININ HA / AIV HAEMAGGLUTININ NEURAMINIDASE HN / NDV NEURAMINIDASE N RNA /AIV 34-00
36 The criminals at work NDV & AIV Attachment Penetration HAEMAGGLUTININ NEURAMINIDASE (HN) + FUSION (F) 35-00
37 The criminals at work NDV & AIV Attachment Penetration HAEMAGGLUTININ NEURAMINIDASE (HN) + FUSION (F) NEURAMINIDASE (N) + HAEMAGGLUTININ (HA) 36-00
38 The criminals at work NDV & AIV Attachment Penetration HAEMAGGLUTININ NEURAMINIDASE (HN) + FUSION (F) NEURAMINIDASE (N) + HAEMAGGLUTININ (HA) To be active, the F0, the precursor of the F protein of NDV as well as HA0, the precursor of the HA protein of AIV need to be cleaved by a protease 37-00
39 The criminals at work NDV & AIV HA
40 The criminals at work NDV & AIV - F0 of velogenic NDV strains, as well as HA0 of Highly Pathogenic AIV strains can be cleaved by many proteases : systemic infection - F0 of lentogenic strains, as well as HA0 of Low Pathogenic AIV strains, can be cleaved only by trypsin- like protease present in a limited number of tissues : local infection 39-00
41 The criminals at work NDV & AIV The cleavage of the F0 or the HA0 is dependant on the amino acid sequence at the level of the cleavage site of the F / HA gene. This sequence can explain, and be used to predict the virulence of an NDV or an AIV 40-00
42 F The criminals at work NDV & AIV (Fusion protein) is the most important factor of virulence: Introduction of an amino-acid sequence of a velogenic NDV inside the cleavage site of the fusion protein F of an avirulent NDV (ICPI = 0) turned it into velogenic (ICPI = 1,28) : GGRQGR L GRRQRR F (Ben Peeters et al., J. of Virology, (1999), ) 41-00
43 HA The criminals at work NDV & AIV (Haemagglutinin) is the most important factor of virulence: Ex : Italy : epizotic of H7N1 AIV : LPAIV (PEIPKGR*GLF) (03/ /1999) IVPI = 0.00 HPAIV (PEIPKGSRVRR*GLF) (12/99-04/2000) IVPI =
44 HN The criminals at work NDV (Haemagglutinin Neuraminidase) Is responsible for the attachment of NDV to the host cells. It is not a factor of virulence : Introduction of an HN gene from a velogenic NDV into a mesogenic NDV (Anhinga virus) failed to increase its virulence. (Estevez C. et al., Virus Research, 129 (2007), ) 43-00
45 NP, M, P The criminals at work NDV (Nucleo / Matrix / Phospho - proteins) Are not important for protection. Serum containing antibodies against NP, M and P proteins did not induce any protection (Reynolds & Maraqa, Av. Diseases, 44, (2000), ) 44-00
46 CONCLUSIONS NDV & AIV The F and HA antigens are the key factors of virulence of NDV and AIV. They are also the key protective antigens. Vaccine protection against ND / AI requires immunity against F / HA 45-00
47 ARE THESE TWO CHALLENGING VIRUSES STABLES OR CHANGING? 46-00
48 Are NDV and AIV changing viruses? NDV AIV 1 Serotype 16 HA & 9 NA = «144» combinations Variations in : - Virulence High / Low pathogenic - Pathotype / tropism + Genotype variations 47-00
49 NDV : a challenging, but stable virus Classification of NDV strains according to virulence Virus ICPI IVPI MDT Ulster 2C to > 168 PHY.LMV to >168 Hitchner B F La Sota ,7 Komarov Roakin Mukteswar Beaudette C GB Texas Essex Herts New York Parrot Milano Pigeon / England / 617 / > 120 Chicken / England / 702 / VIRULENCE APATHOGENIC LENTOGENIC MESOGENIC NDV VELOGENIC NDV (After Heffels-Redmann U., 1992 and Ceva) 48-00
50 AIV : a challenging and changing virus LPAIV (PEIPKGR*GLF) IVPI = 0.00 HPAIV (PEIPKGSRVRR*GLF) IVPI =
51 NDV : a challenging, but stable virus Genetic tree of NDV strains Genogroups
52 AIV : a challenging and changing virus Distance 0.02 Ck/Zhengzhou/1/02 HVT AI insert Gs/GD/1/96 Dk/Guangxi/50/01 IDN/CDC940/06 IDN/CDC836/06 IDN/CDC759/06 IDN/CDC669/06 IDN/CDC739/06 IDN/CDC835/06 IDN/CDC610/06 Ck/IDN/Bandung /06 Ck/West Java/ Subag-29/2007 challenge strain MDk/Jakarta/HABWIN/06 Qa/Jakarta/JU1/06 Ck/West Java/Tasiko/06 Ck/IDN/Sragen /06 Ck/IDN/Garut /06 IDN/CDC699/06 Ck/IDN/Yogyokarta /06 IDN/CDC523/06 Qa/IDN/Bantul /06 Genetic tree of some H5N1 HPAIV strains Clades Ck/ SmiWN18/2009 Ck/Sumatra/D1614-2/2011 Ck/Central Java/D1614-3/2011 IDN/CDC1031/07 IDN/CDC887/06 IDN/CDC938/06 IDN/CDC1032/07 IDN/CDC1047/07 IDN/CDC1046/07 IDN/CDC582/06 IDN/CDC634/06 IDN/CDC644/06 Ck/IDN/Tegal /06 Ck/IDN/Lampung /06 Ck/IDN/Demak /07 Ck/IDN/Jemban /06 Ck/IDN/Probolinga /07 Ck/IDN/Semerang /07 62/07 Ck/IDN/Pekalonguru /07 Swan/IDN/Malang /07 Ck/IDN/ /07 Ck/Jatim/LYR-HW/07 Ck/HW/K-GK/07 Ck/Gunung Kidul/BBVW/07 IDN/CDC370/06 IDN/CDC390/06 Ck/West Java/SMI-ENDRI1/06 Ck/West Java/SMI-ENDRI2/06 Java Distance 0.02 Phylogenetic tree constructed based on entire nt sequence of H gene Reference: Takano et. al. Virology :
53 THE DIFFICULT IMPLEMENTATION OF AN EFFICACIOUS VACCINATION PROGRAM 52-00
54 The problems to vaccinate against ND & AI NDV AIV 53-00
55 The problems to vaccinate against ND & AI NDV AIV Interference with MDA 54-00
56 The problems to vaccinate against ND & AI Antibody titre* (10 log) 5 Passive immunity in poultry serum tears 1 < *ImmunoPeroxydase Monolayer Assay Age (days) (after P.H. Russel, ) 00
57 The problems to vaccinate against ND & AI Interference between MDA and Live ND vaccine % protection / challenge Chickens Vaccinated Control M.D.A. without Age (days) 56-00
58 The problems to vaccinate against ND & AI Interference between MDA and Live ND vaccine % protection / challenge Chickens Vaccinated Control M.D.A. with without Age (days) 57-00
59 The problems to vaccinate against ND & AI Interference between MDA and Live ND vaccine % protection / challenge Chickens Vaccinated Control M.D.A. with without Age (days) 58-00
60 The problems to vaccinate against ND & AI Interference between MDA & killed ND vaccine % of protectio on Killed vaccine - No MDA Killed vaccine - MDA Age at challenge (days) (after Bennejean G. et al., 1978
61 The problems to vaccinate against ND & AI Interference between MDA & killed AI vaccine 7 ANTIBODY RESPONSE TO AI VACCINATION H5N2 INACTIVATED VACCINE HI Titre - Log D1-0,3 D1-0,5 D7-0,3 D7-0,5 No Vacc Age in Days 60-00
62 The problems to vaccinate against ND & AI NDV AIV Interference with MDA Poor quality of vaccine application 61-00
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65 The problems to vaccinate against ND & AI NDV AIV Interference with MDA Poor quality of vaccine application Observance / vaccination 64-00
66 The problems to vaccinate against ND & AI Tropisms of live ND vaccines Tracheopic HB1, La Sota, Clone 30, etc
67 The problems to vaccinate against ND & AI Tropisms of live ND vaccines Tracheopic HB1, La Sota, Clone 30, etc. Enterotropic Phy.LMV 42, VG/GA, Q.V4, Ulster 2C, etc
68 The problems to vaccinate against ND & AI NDV AIV Interference with MDA Poor quality of vaccine application Observance Antigenic / vaccination variations 66-00
69 The problems to vaccinate against ND & AI The limits of killed ND & AI vaccines ND Vaccine Challenge C. Broiler VTM strain ND K HVT-NDV Herts 33 (Gen IV) Malaysian (Gen. VII) 67-00
70 The problems to vaccinate against ND & AI The limits of killed ND & AI vaccines ND Vaccine Challenge C. Broiler VTM strain ND K HVT-NDV Herts 33 (Gen IV) 100% Malaysian (Gen. VII) 100% 68-00
71 The problems to vaccinate against ND & AI The limits of killed ND & AI vaccines ND Vaccine Challenge C. Broiler VTM strain ND K HVT-NDV Herts 33 (Gen IV) 100% 100% Malaysian (Gen. VII) 100% 100% 69-00
72 The problems to vaccinate against ND & AI The limits of killed ND & AI vaccines ND Vaccine AI Vaccine Challenge C. Broiler VTM strain ND K HVT-NDV Challenge strain H5N2 K VTM HVT-AIV Herts 33 (Gen IV) 100% 100% Egypt 2007 Malaysian (Gen. VII) 100% 100% Egypt
73 The problems to vaccinate against ND & AI The limits of killed ND & AI vaccines ND Vaccine AI Vaccine Challenge C. Broiler VTM strain ND K HVT-NDV Challenge strain H5N2 K VTM HVT-AIV Herts 33 (Gen IV) 100% 100% Egypt % Malaysian (Gen. VII) 100% 100% Egypt % 71-00
74 The problems to vaccinate against ND & AI The limits of killed ND & AI vaccines ND Vaccine AI Vaccine Challenge C. Broiler VTM strain ND K HVT-NDV Challenge strain H5N2 K VTM HVT-AIV Herts 33 (Gen IV) 100% 100% Egypt % 100% Malaysian (Gen. VII) 100% 100% Egypt % 100% 72-00
75 Summary of challenge experiments conducted with Vectormune HVT AIV Type Clade Country Year Strain ref. H5N1 Protection % 1 VIETNAM 2004 A/Vietnam/1203/ % 2.2 HUNGARY 2006 EGYPT A/duck/Hungary/11804/ % A/chicken/Egypt/ VIR08/ % A/chicken/Egypt/1709-6/ % 2010 A/chicken/Egypt/63/ % H5N INDONESIA NA A/chicken/WJava/Subang/0 29/ % A/Ck/Purwakartacilingga/142/ % MEXICO 1995 A/Chicken/Queretaro/ % USA 2004 A/Parrot/US/ %
76 THE CONCEPT OF VECTOR VACCINATION 74-00
77 The Concept of Vector Vaccination % protection / challenge MDA Age (days) 75-00
78 The Concept of Vector Vaccination % protection / challenge MDA Vectormune HVT-NDV Age (days) 76-00
79 The Concept of Vector Vaccination % protection / challenge MDA Vectormune HVT-NDV Cevac Vitapest L Age (days) 77-00
80 The Concept of Vector Vaccination % protection / challenge MDA Vectormune HVT-NDV Cevac Vitapest L Vectormune + Vitapest Age (days) 78-00
81 The Concept of Vector Vaccination Antibody Response to Vectormune HVT-NDV and Protection. 7 6 Vectormune HVT-NDV - 1 Controls Log2 HI titres Age in Days 79-00
82 The Concept of Vector Vaccination Antibody Response to Vectormune HVT-NDV and Protection. 7 6 Vectormune HVT-NDV - 1 Controls % 5 Log2 HI titres % 68% 95% 2 1 5% 0% 0% 0% Age in Days 80-00
83 The Concept of Vector Vaccination Antibody Response to Vectormune HVT-NDV. 7 6 Vectormune HVT-NDV - 2 Controls Log2 HI titres Age in Days 81-00
84 The Concept of Vector Vaccination Clinical protection of broilers over time (3, 4, 5, and 6 wks of age) after NDV Chimalhuacan mucosal challenge Ch1 120 (3 wks) Ch2 (4 wks) Ch3 (5 wks) Ch4 (6 wks) protectio on % Clinical rhvt/nd rhvt/nd+live ND live ND broiler control Group 1 Group 2 Group 3 Group
85 VECTORMUNE HVT-NDV Onset of Immunity Spectrum of Immunity Duration of Immunity Reduction of shedding 74-00
86 VECTORMUNE HVT-NDV Onset of Immunity Spectrum of Immunity Duration of Immunity Reduction of shedding 74-00
87 Vectormune HVT-NDV Antibody Response to Vectormune HVT-NDV - Comparison with another rhvt-ndv. 7 6 Vectormune HVT-NDV - 1 rhvt-ndv Controls Log2 HI titres Age in Days (Ceva results 2012) 18-00
88 Vectormune HVT-NDV Antibody Response to Vectormune HVT-NDV - Comparison with another rhvt-ndv. 7 6 Vectormune HVT-NDV - 1 rhvt-ndv Controls Log2 HI titres Age in Days (Ceva results 2012) 18-00
89 Vectormune HVT-NDV Protection from Vectormune HVT-NDV - Comparison with another rhvt-ndv rhvt-ndv Vectormune HVT-NDV - 1 Controls rotection Rate 60 P Age at Challenge (days) (Ceva results 2012) 19-00
90 VECTORMUNE HVT-NDV Onset of Immunity Spectrum of Immunity Duration of Immunity Reduction of shedding 74-00
91 Vectormune HVT-NDV Does it protect against various types of NDV? F insert
92 Vectormune HVT-NDV Does it protect against various types of NDV? F insert Yes!
93 Vectormune HVT-NDV Does it protect against various types of NDV? Yes! F insert Yes!
94 Vectormune HVT-NDV Does it protect against various types of NDV? Yes! Yes! F insert Yes!
95 Vectormune HVT-NDV Does it protect against various types of NDV? Yes! Yes! Yes! F insert Yes!
96 VECTORMUNE HVT-NDV Onset of Immunity Spectrum of Immunity Duration of Immunity Reduction of shedding 74-00
97 Vectormune HVT-NDV Controlled trials in Hungary - Layers - Lohman Brown layer pullets with MDA (mean HI = 5,5 log 2) - Vectormune HVT-NDV (d1 SQ) - Vectormune HVT-NDV (d1 SQ) + Cevac Vitapest L (ED) - Cevac ND IB EDS K (w15 IM) - Challenge : EID 50 Malaysian 2010 vvndv viscerotropic isolate (Genotype VII) - IN route (0,1 ml) - at 3, 4, 6, 10, 15 and 33 weeks of age 83-00
98 Vectormune HVT-NDV Protection o of layers against vvndv challenge % Protecti ion Vectormune HVT-NDV Vectormune HVT-NDV + Cevac Vitapest L + Killed Controls Age at challenge (in weeks) 84-00
99 Vectormune HVT-NDV Challenge at 33 weeks: egg production ND challenge (w33) * *negative controls not challenged 85-00
100 VECTORMUNE HVT-NDV Onset of Immunity Spectrum of Immunity Duration of Immunity Reduction of shedding 74-00
101 Vectormune HVT-NDV DISEASE? 1 Infection 2 - Clinical expression 53-00
102 Vectormune HVT-NDV DISEASE? 1 Infection 2 - Clinical expression 3 Re-excretion 54-00
103 Vectormune HVT-NDV PROTECTION? 1 Infection 2 - Clinical expression CLINICAL PROTECTION 3 Re-excretion 55-00
104 Vectormune HVT-NDV PROTECTION? 1 Infection 2 - Clinical expression CLINICAL PROTECTION 3 Re-excretion REDUCTION OF RE-EXCRETION 56-00
105 Vectormune HVT-NDV PROTECTION? 1 Infection HIGHER RESISTANCE TO INFECTION 2 - Clinical expression CLINICAL PROTECTION 3 Re-excretion REDUCTION OF RE-EXCRETION 57-00
106 Vectormune HVT-NDV Trials at SSIU / Ceva Phylaxia - Commercial broilers (20 per group) - Vectormune HVT-NDV SQ or in-ovo (-3d) - Challenge : EID50 Chimalhuacan NDV strain - at 3 or 4 or 6 weeks of age - IN + ON route - Oropharyngeal and Cloacal swabs taken 3 and 7 days post challenge - Virus quantification using RT-PCR 58-00
107 Vectormune HVT-NDV Clinical protection
108 Vectormune HVT-NDV Challenge at D28 * * * * * * * * Reduction of shedding 60-00
109 Vectormune HVT-NDV Challenge at D40 * * * * * * * * Reduction of shedding 61-00
110 Vectormune HVT-NDV / Vectormune HVT-AIV CONCLUSIONS Regarding vaccine prevention of ND and AI, Vectormune HVT-NDV and Vectormune HVT-AIV are more revolutions than just evolutions. Compared to classical vaccines / programs, the protection induced is : -simpler - stronger - more reliable - deeper - and longer lasting 86-00
111 Vectormune HVT-NDV / Vectormune HVT-AIV CONCLUSIONS They actually reduce the shedding of the field virus after challenge, and consequently the risk of spreading. For this reason, Vectormune HVT-NDV and Vectormune HVT-AIV do open the door to a real control of the 2 disease. Both of them require injection at day old or in- ovo. For this reason, all efforts should be made to ensure a perfect application at the level of the hatchery 87-00
112 Vectormune HVT-NDV / Vectormune HVT-AIV How to solve ND & AI vaccination problems? vaccines : RESISTANT to MDA APPLICABLE AT DAY 1 BROAD SPECTRUM and SAFE 88-00
113 Vectormune HVT-NDV / Vectormune HVT-AIV 89-00
114 Vectormune HVT-NDV / Vectormune HVT-AIV THANK YOU FOR YOUR ATTENTION 90-00
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