Development of Infectious Bronchitis Variant 2 vaccines from

Development of Infectious Bronchitis Variant 2 vaccines from a local Israeli isolate as a solution for Unusual virulent field strain Udi Ashash, St. Petersburg 09 2014

Distribution of Israeli IB Variants 1996 -2011 100 90 % of isolates 80 70 60 50 40 30 20 10 0 1996 1997 1998 1999 2000 2001 2002 2003 2004 2005 2006 2007 2008 2009 2010 2011 Mass 793 B IS/720 IS/885 QX-like IS/1201 Variant 2 Data obtained from Dr. Rozie Meir, Kimmron Veterinary Institute, Israel

Pathological Findings following IB Variant 2 (IS/1494/06) field Infection (Dr B. Perelman, Dr K. Elrom) Tracheal plugs Enlarged swollen kidneys Severe and hyper acute airsaculitis Watery cyst in reproductive tract

GQ 154655 IBV H-120 95% MASS DQ 664534 IBV Massachusetts 41 EU 283057 Conn/Bvial 1 98% Phylogenetic Tree of IB field and vaccines strains based on S 1 gene DQ 400359 IS/1201 FJ 807931 QX K 1583 QX FJ 807946 KM 91 79% DQ 459474 YN 05 -01 AF 093793 IBV 4/91 attenuated 95% AF 093794 IBV 4/91 virulent EU 350550 IBV IS/1366 99% 793/B AF 093795 IBV Variant 1 JQ 946056 IB VAR 233 A ABIC AY 279533 IS/885 S 1 98% 83 -88% AY 091552 Israel/720/99 EU 368592 Egypt/Zag/07 -01 Unique Variants IBV IS/2006/08 GQ 281656 Iraq Sul/01/09 X 15832 D 274 83% AY 789950 IBV IS/223/96 AF 093796 IBV variant 2 Approx. 60% 92% IBV IS/373/09 JX 027070 IB VAR 2 -06 ABIC EU 780077 IBV IS/1494/06 S 1 EU 283055 Ark/Cvial 2 AF 274437 GA/0470/98 EU 283063 Del/Cvial 1 AF 274435 DE 072 0. 05 % represents the identity of the S 1 gene Variant 2

• 220 tracheal swabs were taken from 8 flocks of breeder chickens and 2 flocks of broilers chickens • All 8 Turkish isolates sequences were related to IS/1494/06

2 nd Study in Turkey -

Current IB epidemiology in Turkey

IS/1494/06 (Var 2) Middle east epidemiology ? ? ? http: //phibrovaccinesmap. com

History of IB vaccines in Israel In the last 15 years the Massachusetts strain vaccine did not protect adequately Since 2007, the use of 793 B vaccine strain reduced the occurrence of some variants but not the IS/1494/06 IB variant 2 (IS/1494/06) became prevalent and caused high mortality and severe economic losses The need for a new homologous vaccine became a must requirement

Development of “TAbic IB VAR 206” Vaccine A live-attenuated vaccine originated from IB Variant 2 isolated in Israel (IS/1494/06) was developed The vaccine was tested under laboratory and field conditions At this stage “IB VAR 206” vaccine is currently in extensive use under field conditions in broiler chickens in Israel and Turkey

Safety tests of “TAbic IB VAR 206” in Broilers Vaccination age (days) Vaccination administration route 1 Coarse spray 1 Aerosol spray 1 Eye drop 1+12 Dosage Morbidity and Mortality post vaccination At least 10 doses (105. 4) At least 10 doses (105. 2) 0/12 Coarse spray 1 dose (104. 4) 0/12 1+12 Aerosol spray 1 dose (104. 4) 0/12 1+12 Eye drop 1 dose (104. 2) 0/12 No vaccination 0/12

Protection from challenge in SPF chickens vaccinated at 1 and 14 days of age with “TAbic IB VAR 206” vaccine and challenged with homologous Variant 2 (IS/1494/06) 97. 3 100 92 IB VAR 206 vaccination at 1 and 14 days of age Not-vaccinated and challenged 14 0 % Cilia motility % protection, negative swabs Not-vaccinated and notchallenged

2 IB VAR 206 vaccinations provide better protection against virulent challenge compared to combinations of H-120 & 793 B 97 a % Cilia motility H-120 (1) and 793 B (14) 67 b IB VAR 206 (1 and 14) Not vacc. & challenged 0 c Challenge with VAR 2 field strain IS/1494/06

Interference test following TAbic IB VAR 206 and NDV simultaneous vaccination Antibodies level, NDV HI (log 2) 6. 4 a 6. 6 a Protection against NDV challenge 100 a 90 a Protection against IBV challenge 86 a 2 b 0 b NDV vaccine NDV and IB VAR 2 -06 Not vaccinated NDV vaccine NDV and IB VAR 206 Not vaccinated 7 b IB vaccine NDV and IB VAR 206 Not vaccinated There is no interference between the two viruses

IB Vaccination protocols for broilers used in Israel Vaccination with TAbic IB VAR 206 vaccine at 1 and 12 -14 days of age Or Combined vaccination of TAbic IB VAR (793 B) and TAbic IB VAR 206 at 1 and 12 -14 days of age

2. 0% 2002 -1 2002 -4 2003 -1 2003 -4 2004 -1 2004 -4 2005 -1 2005 -4 2006 -1 2006 -4 2007 -1 2007 -4 2008 -1 2008 -4 2009 -1 2009 -4 2010 -1 2010 -4 2011 -1 2011 -4 2012 -1 2012 -4 2013 -1 Mortality in an Israeli Chicken integration 2002 -2013 14. 0% 12. 0% 10. 0% 8. 0% 6. 0% 4. 0% I II 0. 0%

/14 IS 06 79 3 B AR R 2 Mortality % VA V 6 4 IB 9 IB 11 c bi TA bi c 10 8 94 /06 12 TA Mortality trend in the same Israeli chicken integration in relation to the introduction of variant IB vaccines 7 5 5 2 0 2004 2007 2008 2009 2012

Turkey – Improved performance after using “TAbic ® IB VAR 206” Heavy losses (up to 12% average mortality) in broiler flocks in integrations across Turkey TAbic IB VAR 206 was introduced in April 2013 Up to now reports of reduction in respiratory signs and secondary infections 4 -7% reduction in mortality Improved FCR, body weight and reduction in slaughter house condemnations No interference with other vaccines (ND, Gumboro)

IB variant 2 (IS/1494/06) in breeders and layers Infection with IB Variant 2 (IS/1494/06) at early age causes permanent damage to the reproductive tract Infection during the laying period causes reduction in egg production and egg quality There is a need for an inactivated IB Variant 2 (IS/1494/06) vaccine as an important tool for the protection of breeders and layers against field exposure and possible maternal protection

Protection Following Inactivated Vaccination Inactivated IB Variant 2 vaccine induces high levels of specific antibodies and provides protection against challenge under the laboratory conditions Vaccinations Antibodies Titer Protection 14 days 42 days 4 w post 1 st vaccination TAbic IB VAR 206 Inactivated IB VARIANT 2 445 3040 95% Inactivated IB VARIANT 2 176 1557 90% 1. 6 26. 3 90% Not-Vaccinated, not-challenged Not-Vaccinated and Challenged 5 w post 2 nd vaccination % Cilia Motility 0%

Summary Variant 2 field strain is currently the main IB strain found in the Middle East, Gulf states, North Africa and Turkey with a capacity to go global TAbic ® ‘IB VAR 206’ is the only available homologous tablet vaccine against IB Variant 2 field strain TAbic ® ‘IB VAR 206’ is safe for Broiler chickens from day one TAbic ® ‘IB VAR 206’ shows high efficacy in both lab trials and field use

Summary Mass and 793 B vaccines failed to protect chickens in both Israel and Turkey Two vaccinations of TAbic IB VAR 206 at 1 and 14 days of age were found to provide adequate protection against field exposure Inactivated IB Variant 2 proved to be protective against challenge and will be an important tool in the protection of laying production in Breeders and Layers

Thanks to the Phibro Team Tal Even Chen M. S. C Sarit Heifez M. S. C Dr. Beny Perelman Dr. Eitan Kin Dr. Caroline Banet-Noach Dr. Avner Finger Tiran Ella

Thank you!