Newcastle Disease Dr. - PowerPoint Presentation

1. Newcastle Disease Dr. Hatem S. Abd El-HamidProf. of Poultry Diseases & Hygiene Fac. Vet. Med. Former of Damanhour Univ., Egypt30/1/2017

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3. Welcome in EGYPT

4. Be Relax in Sharm El-Sheikh

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11. Newcastle Disease Virus (NDV)Order : mononegivirals Family : paramyxovirida Sub family : paramyxovirinae Genus : Avulavirus Serotypes: APMV-1(APMV-1 to APMV-19)Genetic analysis classify NDV to class I, class IIClass I genotype avirulent strains. Class II genotype both virulent and avirulent.

12. Snoeck et al. (2013) NDV genotypes (I- XVIII)Genotype IGenotype IIGenotype IIIGenotype IVGenotype VGenotype VIGenotype VIIVII AVII BVII CVII DVII EGenotype VIIIGenotype IXGenotype XGenotype XIGenotype XIIGENOTYPE XIIIGenotype XVGenotype XVIIGenotype XVIIIClass IIGenotype XVIGenotype IVX

13. Distribution of NDV Genotype VII ●Kazakhstan, Genotype VIIb, VIId●Japan, Genotype VIIdKoreaGenotype VIId●●China, Genotype VIIa, VIIb, VIIc, VIIdTaiwan, Genotype VIIc, VIId, VIIe●●IndonesiaGenotype VIIa●MalaysiaGenotypeVIIa, VIIb, VIIc, VIId●IndiaGenotypeVIIb●NepalGenotypeVIIb●R.S.A.GenotypeVIId●Libya, Genotype VII●Turkey Genotype VIIb●venezuelaGenotypeVIIb●EgyptGenotypeVIId

14. NDV World PanazooticsGenotype I……………………… a virulant Genotype II,III,IV…………. 1st panazootic (started in southeast Asia in the mid-1920) Genotype V ……………………..2nd panazootic (started in the Middle East in the late 1960s and to have spread to most countries by the early 1970)Genotype VI…………….ppmv-3rd panazootic ( in the Middle East in the late 1970, spread to Europe in the 1980), Genotype VII and VIII ……. 4th panazootic (since the early 1990s to current date in Asia, Europe and Africa including Egypt )new sub-genotypes of vNDV suggests that a new, fifth, panzootic of ND already originated in Southeast Asia, extended to the Middle East, and was entering into Eastern Europe. (Miller et al., 2015)

15. Molecular characterization of the sequenced isolates of NDVSerial NumberAbbreviationGenotypeHostVirulenceCleavage site of F protein112RRQKR116F117Gene Bank Accession No18NDV/EG/CH/18 /2015VII dChickenvirulentRRQKRFKu37778126NDV/EG/CH/26/2014VII dChickenvirulentRRQKRFKu37778335NDV/EG/CH/35/2014VII dChickenvirulentRRQKRFKu37778424NDV/EG/CH/24/2014VII dChickenvirulentRRQKRFKu377782

16. Gene Bank data of NDV isolates used in comparative analysis with the 4 Egyptian NDV isolates in this study. StrainCountryAbbreviationHostAccession NumberGenotypeLasota LasotaChickenAF077761IIHitchinerUSAHITCHINER BIChickenJN872151IIAvinew VG/GAUSAVG/GAChickenEU289028IIVECTORMUNE HVT ND D26/76ChickenM24692IVITAPEST PHY-LMV42ChickenDQ097394ICLONE 30  ChickenY18898 IINDV/Chicken/2/2006Egypt93/EG/06ChickenFJ969393IINDV/Chicken/ 3/2006Egypt94/EG/06ChickenFJ969394IINDV/Chicken/4/2006,Egypt95/EG/06ChickenFJ969395IINDV/chicken/VRLCU138/ /2012Egypt68/EG/12ChickenJX885868VIIdNDV//CH/ SDWF07/2011CHINA ChickenJQ015295/KF935230VIId

17. Amino acid identity of 4 NDV isolates with NDV strains and NDV vaccines

18. Phylogenetic tree for the 4 NDV isolates and related NDV strains based on the fusion protein amino acids sequences.

19. Avian Paramyxovirus – 1 (APMV-1) Order: Mononegavirales Family: Paramyxoviridae Sub-Family: Paramyxovirinae Genera: Avulavirus Serogroups: 19 Serogroups (APMV 1 –19) (PPMV-1): Pigeons & Doves - Antigenic variant of NDVHNL, NP, P (V)Newcastle Disease Virus

20. NP: NucleoproteinP: Phosphorylated protein (Overlapping V gene protein)M: MatrixF: Fusion with host cell membrane (Smaller spike)HN: Hemagglutinin / Neuraminidase activities (Largest spike)L gene: RNA-Directed RNA polimerase (Nucleocapside)NPPMF0HNLGenomic FeaturesEnveloped pleomorphic RNA virus100-500 nm sizeSingle-stranded; Non-segmentedNegative sense (15kb) Helical nucleocapside symmetry(Herring-bone like structure)NDV GenomeFeaturesLentogenic112RRQRRF117Velogenic

21. NDV was recoded in Egypt for the 1st time during 1942. Outbreaks causing significant losses in the poultry industry since 1948. Since that time , isolation of velogenic NDV was successflly carried out by many authors .

22. Between (2004 – 2006 )STUDIES ON NEWCASTLE DISEASE AT SHARKIA GAVERNORATE WITH SPECIAL REFRENCE TO CHARACTERIZATION AND DIAGNOSIS.100 poultry farms were examined for NDV.The incidence of the viruses infection was 24% 17 isolates was Velogenic , while 7 isolates were characterized as lentogenic isolates(MDT) The infection was high during cold seasons .

23. During 2005CHARACTERIZATION OF VARIANT STRAIN OFNEWCASTLE DISEASE VIRUS During 2005, the virus was isolated from broiler farm suffered from more than 95% mortalities.The isolates confirmed to be velogenic viscerotropic NDV by mean death time (MDT)Results indicated that the live vaccine did not protect against morbidity but reduced mortality

24. ND virus strains isolated during 2005 & 2006 were probably circulated in vaccinated flocks.Developed an outbreaks were closely related to genotype II of class II NVD.

25. Pathotyping using sequence gene analysis carried out by 2005.In Egypt, since 1970s till 2009, most of NDV isolated was found belong to genotype VI (which endemic in Egypt).

26. SEQUENCE ANALYSIS OF FUSION PROTEIN GENE OF NEWCASTLE DISEASE VIRUS ISOLATES FROM OUTBREAKS IN EGYPT DURING 2006The sequence of fusion protein (F) gene of the three (NDV) strains isolated from outbreak in chicken showed that the Egypt isolates are closely related with the genotype II of class II NDV strains.

27. During 2009 COMPLETE GENOME SEQUENCE OF A VIRULENT NEWCASTLE DISEASE VIRUS ISOLATED FROM AN OUTBREAK IN CHICKENS IN EGYPT The phylogenetic analysis showed that Egyptian strain Egypt is closely related with the NDV strains isolated in China, the sequence of the fusion protein cleavage site of Egyptian stains was identical to that of the NDV strain recently isolated in Mali.

28. During 2011MOLECULAR CHARACTERIZATION OF NEWCASTLE DISEASE VIRUS GENOTYPE VIID IN INFECTED BROILER FLOCKS IN EGYPTIn early months of 2011 sever outbreaks of NDV affecting the commercial poultry was reported.The NDV isolate was found velogenic in nature.Phylogenetic analysis revealed that the isolated NDV is belonging to sub- genotype VIId and in close range to other Chinese VIId isolates based on the high nucleotide similarity between them which indicate the genotype VIId is circulating between chicken flocks.

29. Between 2011 - 2012ISOLATION AND MOLECULAR CHARACTRIZATION OF NEWCASTLE DISEASE VIRUS IN EGYPT BETWEEN 2011 AND 2012 NDV was isolated from Ten different vaccinated broiler farms in Fayom , Behira and Giza Provinces .In THREE positive samples , ONE isolate was velogenic isolate class II genotype VII sub genotype d NDVs and TWO isolates class II genotype II NDVs .

30. Between 2011-2012 (South valley)MOLECULAR CHARACTERIZATION OF NDV ISOLATED FROM CHICKEN IN SOUTH VALLERY PROVINCES ( Sohag, Qena and luxor) This Study was carried out on 126 samples.The obtained results indicated that 45 samples(35.7%) were NDV positive.Characterization of NDV isolated revealed . Mesogenic (2/45) , velogenic (43/45) type.( Sohag, Qena and luxor)

31. Phylogentic Analysis indicate the presence of new NDV strains differ from strains of 2006 and the vaccinal strains used for vaccination.Studies on comparison of antigenic structure of the newly pathogenic NDV strains isolated from different outreaks with the viruses used for the vaccination are of crucial importance to understand the causes of vaccination failure

32. During 2012MOLECULAR CHARACTERIZATION OF FIELAD AND VACCINAL STRAINS OF NDV WAS CARRIED OUT :The old NDV genotype VI which used to be circulating in Egypt since 1970s, and the new NDV genotype VIId, currently circulating in Egypt. One of the isolates , which was isolated in 2012 from Giza, was found to belonging to genotype VIIdOne of the isolates, which was isolated in 1998, was belong to genotype VI, which endemic in Egypt since 1970s till 2009.Four isolates found to be vaccine strains .

33. Result of Survey of poultry Flocks During (January-May,2013)Total number of examined flocks=204.The examined flocks represent 16 governorates as following:Upper Egypt: El-fayom, Beni-suif, El-menia, Assuit, Sohag and QinaEl-Giza, el-kaloubia, El-monofia, El-Gharbia, EL-beheira, Alexandria, Matrouh, El-sharkia, Ismalia and El-Suez.The Flocks (204) were Invastigated for the Following diseases:AIV (H5 & H9)NDVIBV

34. PCR Finding and Analysis for 204 Examined FlocksViral agentsNO.of cases % of Findings out of 204% of IBV cases out of 149 % of H9 cases out of 54% of NDV out of 49H56 3 %---H912 23 % -22 %-NDV74 % --14%IBV7336 % 49%--IBV + NDV2914 %19.5 %-59% IBV+NDV+H91121 %7.4 %20%22%IBV + H92955 %19.5 %54 %-IBV + H573.5%5 %--NDV+H924 %-4%4%Negative cases2613 %---Total Number of Examined cases2041495449

35. During 2014STUDIES ON ISOLATION AND CHARACTERIZATION OF NDV IN BROILER AND LAYER CHICKEN FLOCKS IN EGYPT REVEALED :IN 195 broiler and layers farms from eighteen Egyptian governorates 24% was positive for virulent NDV. Sequence analysis for F gene for twelve NDV isolates revealed that they are of virulent type and genetically related to genotype VII NDV except 2 isolates are virulent type resemble lasota life strain

36. DETECTION OF NVD WAS CARRIED OUT ON 27 ALLANTOIC FLUID SAMPLES & 26 CLINICL SWAB SAMPLESThe results revealed the presence of NDV in 11 samples. Pathotyping using MDT characterized Virulence of Nine isolates of MDT less than 60 hrs which were typical to Virulent strains of NDV and 2 isolates of MDT more than 90 hrs which are typical to avirulent type NDV .Phylogentic analysis proved that 8 isolates of genotype VI and remaining 3 isolates of genotype II which indicating that most of Egyptian isolates circulating in Egypt are of these TWO GENOTYPE.During 2014

37. During 2014MOLECULAR CHARACTERIZATION OF NEWCASTLE DISEASE VIRUSES IN CHICKEN DURING 1996-2009Characterization of some (NDV) isolates from different localities and different year in Egypt carried out and 11 NDV Out of 53 positive samples for NDV were characterized .9 isolates were of virulent type with MDT less than 60 hrs and TWO isolates showed MDT more than 90 hrs.(Avirulent).Partial fusion gene sequence revealed revealed that EIGHT isolates are close to genotype VI and THREE isolates are close to genotype II.

38. Mohamed ET AL.(2011) ND virus strains isolated during 2005 & 2006 were probably circulated in vaccinated flocks.Developed an outbreaks were closely related to genotype II of class II NVD.

39. Ismail et al.(2014)2011-201355 NDV isolates velogenic strains.9 isolates exposed to sequencing belong to genotype VII d . 

40. During 2014 -201648 broiler,commercial layer and broiler breeder flocks 4 Governorates (Gharbia, Behera,Dakahlia, Kafer El-Shiekh) suffered from high mortality were examined for NDV. 21 HA positive samples out of 48 flocks were recroded . 6 out of 21 HA positive samples were NDV (28.5%).

41. Molecular characterization of 4 sequenced NDV isolates indicated that the current isolated strains belonged to genotype VII sub genotype d which is predominant genotype responsible for most ND outbreak since the end of the last century and currently circulating in Egypt

42. ConclusionEgypt is endemic for NDV with continuous long lasting outbreaks causing sever economic losses in poultry industry.Most circulating Egyptian NDVs are in close range to genotypes II and VI during 1970s till 2009.Since,2011 to 2017 genotype VII NDV strains of class II have maintained a constant threat for commercial poultry farm in Egypt.

43. Conclusion continue..Presence of new NDV from which differ from the vaccine strains used for vaccination programs in Egypt.5. Comparison of antigenic structure of newly NDV strains isolated from outbreaks with that of the virus used for the vaccination are of crucial importance to understand the causes of frequently occurring vaccine failure.

44. Thank You