1. Dr. Abdul Rehman Poultry Research Institute, Rawalpindi

2. Newcastle disease First identified in Newcastle in 1926 Affects all species of birds Mortality up to 100% in chickens Widespread in Asia, Africa, Europe and South America In USA, sporadic Newcastle disease (ND) occur due to importation of infected birds. In 2002 ND outbreak in California, four million birds were depopulated and cost the U.S. billions of dollars in damage and lost trade. In Pakistan V-virulent ND appeared in Nov,2011

3. Clinical History (NDV) Broilers: Highest economic losses Affected age is 25-35 days, also observed at 8-14 days Layers: Maximum losses in rearing age, Also caused high mortalities & production losses in laying birds Broiler breeders: same as in layers Fancy birds: High mortalities in peacocks, pheasants and pigeons

4. Clinical signs Respiratory distress Greenish whitish diarrhea Torticollis Reduction in production Mortality up to 100%

5. Proventicular hemorrhages. Caecal tonsils hemorrhages Enteritis Tracheitis Postmortem lesions

7. ND Losses in Broilers MonthMortality (Mill)Av. Rate/Kg (Rs)Value (Mill Rs.) Nov,110.86108.0392.90 Dec,113.68117.58432.69 Jan,128.60148.191274.40 Feb,1215.48151.312342.27 Mar,1215.97136.002171.92 TOTAL44.596314.18

8. Population under report at PRI Million MonthBroilerLayerBreederTotal Nov,112.4223.5940.1296.145 Dec,112.3122.8720.1865.369 Jan,122.0692.6890.2394.997 Feb,122.8232.5700.2085.601 Mar,122.7392.4730.2175.233

9. ND Incidence In Punjab

10. Newcastle disease virus Genus Avulavirusin family Paramyxoviridae The genome is a single-stranded negative-sense RNA consisting of 15,186 nucleotides The genome contains six genes in the order of 3’-NP-P-M- F-HN-L-5’ The virus is enveloped, roughly spherical, with a diameter around 100-500nm. F and HN proteins form the external envelope spikes

11. NDV isolates Based on severity of disease, NDV isolates are grouped into three pathotypes: –Lentogenic strains Cause mild or in-apparent respiratory disease –Mesogenic strains Cause respiratory or nervous signs with moderate mortality –Velogenic strains Cause severe intestinal and/or neurologic disease resulting in high mortality –Neurotropic –Viscerotropic

13. Biological characterization of virus Isolation & identification of virus Characterization of virus Mean death time (less than 50 hrs) ICPI (more than 1.5) This indicates it’s a velogenic New castle disease virus Sequencing of M, HN, F genes F gene nucleotide similarity 97 to 100% M gene nucleotide similarity 98.5 to 100% HN gene nucleotide similarity 99.2 to 100% This indicates mutation and close relation with linage 7

14. Contributing factors Indiscriminate vaccination Close proximity of farm Improper biosecurity/ management Improper disposal of dead birds Morbid Bird’s marketing Insufficient disease diagnostic facilities

15. ND Surveillance at PRI (Nov,11 to date) Total no. of sera samples18847 Total no. of tracheal swab 8646 Total no. of cloacal swab 20175 Total no. of tissue sample 2350 Total sample collected 50018

16. Sample processing Embryo inoculation Virus isolation HA HI RT-PCR

17. Results (NDV isolated) Broilers flocks33 Layer flocks 22 Rural flocks 12 Broiler breeder flocks 05 Fancy bird flocks 07 Total isolates 79

18. Research trial on different vaccine regimes Lasota, Mukteswar (live, killed), Velogenic killed (oil based) were employed Mukteswar killed (oil based) and Velogenic killed (oil based) vaccines prepared in disease section PRI, Rawalpindi 7 groups was made 6 experimental and one unvaccinated control

19. Experimental Vaccination schedule Group No. Day of vaccination Day 1Day 10Day 21Day 28 1 ND+IB (E/D)Lasota (E/D)Lasota (D/W)Velogenic field strain challenge 2 ND+IB (E/D)Mukteswer Live (E/D) Lasota (D/W) 3 Mukteswer Killed injection + ND+IB (E/D) Mukteswer Live (E/D) Lasota (D/W) 4 Velogenic Killed injection + ND+IB (E/D) Mukteswer Live (E/D) Lasota (D/W) 5 ND+IB (E/D)Mukteswer Killed injection + Lasota (E/D) Lasota (D/W) 6 ND+IB (E/D)Velogenic Killed injection + Lasota (E/D) Lasota (D/W) 7 No vaccine

21. HI titers log 2 Group No.Age in days Day 1Day 9Day 20Day 27 Group 16.23.33.45.0 Group 25.53.24.44.7 Group 37.33.63.25.5 Group 45.42.73.75.9 Group 56.34.54.46.2 Group 66.23.23.55.6 Group 76.63.42.02.2

22. Protection after experimental challenge Group No.Total no. of birds at 28 days Mortality with in 8 days Total birds survived % protection 143103376.74 242093378.57 34373683.72 44273583.33 54373683.72 64233992.85 7424112.3

24. Results Velogenic killed vaccine at 10 th day along with live lasota vaccine gave maximum protection to 92.85 % Velogenic killed vaccine at day 1 st not proved so well as day 10 th Mukteswer killed vaccine at day 10 th also gave significant protection 83.72 % Mukteswer killed vaccine at day first did not prove as much protective Mukteswer killed vaccine proved well as compared to live vaccine

25. Conclusion Homologous vaccine gave better result as compared to Mukteswer and Lasota vaccines Homologous vaccine may also reduce virus shedding during infection Homologous vaccine along with eradication program may be helpful in the control of current outbreak

26. Research findings of various organizations UVAS Repeatedly isolated and confirmed presence of ND virus in diseased birds Killed vaccine prepared from local isolate gives almost 100% protections against the challenge from the field virus NRLPD Virus is closely related to the Malaysian ND virus isolate Protection study showed that clone killed vaccine prepared from local isolate gave better protection VRI Live and Killed vaccine prepared from Mukteswer proved helpful

27. Recommendations Strict biosecurity rules must be implemented Proper disposal of dead birds Proper dis-infection between the flocks Proper litter disposal It was unanimous opinion that killed vaccine must be included in the vaccine schedule Repeated vaccine should be discouraged (Too many) Vaccine prepared from local isolate should only be used for experimental purposes and should not be used at commercial level More study work needs to be done to find out the solution to this problem Regulatory control in poultry sector Maintain farms proximity Biosecurity/ management Sale/ Disposal of birds Augment Poultry disease diagnostic facilities

28. Thanks