PREVALENCE OF NDV-VII.1.1, LPAI-H9N2 AND HPAI-H5N8 IN CHICKENS IN 2 EGYPTIAN GOVERNORATES DURING LATE 2020
Abstract: Newcastle disease virus (NDV) and Avian Influenza (AIV) incriminated in many respiratory disease problems and egg production decline affecting both broilers and laying hens worldwide. Isolation and identification of NDV, AIV were carried out on 50 diseased chicken (43 broiler & 7 layer) flocks collected from Damietta and Dakahlia Governorates, Egypt. Thirty eight out of 50 examined flocks (76%) showed respiratory signs [37 broiler (74%) and 1-layer (2%) flocks]. The other 12 examined flocks exhibited general signs of illness with greenish diarrhea. Samples from the examined flocks were inoculated into ECE via allantoic sac route and the collected allantoic fluids were tested for hemagglutination activity (HA) which revealed that 21 out of 50 flocks were HA positive (42%); 19 broiler (90.47%) and 2-layer (9.52%) flocks. The twenty-one HA positive samples were subsequently subjected to real-time reverse-transcriptase polymerase-chain-reaction (rRT-PCR). Fifteen samples were positive for Velogenic NDV (71.42%); 3 samples were positive for low pathogenic avian influenza (LPAI) subtype H9 (14.28%); one samples was positive for highly pathogenic avian influenza (HPAI) subtype H5 (4.76%) and 2 samples showed mixed infection for both NDV and H9N2 (9.52%). The Phylogenetic analysis identified the Velogenic NDV genotype VII.1.1 with the amino acid sequence of the three isolates fusion protein cleavage site carries motif 112RRQKRF117. The partial HA gene sequence classified the one selected LPAI-H9N2 isolate as a member of G1 lineage within group B with dibasic amino acid cleavage site motif of 333PARSSR/GLF341 which is distinctive to LPAI. While the other selected isolates of HPAI-H5 belonged to sub type H5N8 clade 22.214.171.124b with multibasic amino acid cleavage site motif of 73PLREKRRKR/GLF84 which is characteristic to HPAI. Our data highlighted the vital role of these viruses in the respiratory disease in poultry flocks. Further continuous studies are necessary for assessing new emerging strains to AIV and NDV and to discuss the impact of the co-infections of these viruses in order to learn more details for their prevention and control.
Key words: Newcastle disease virus, NDV-VII.1.1, Avian Influenza virus, HPAI-H5N8, LPAI-H9N2; chicken flocks
~ 1. Ian B, Paolo M, Krzysztof S, et al. Avian influenza overview October 2016–August2017. J EFSA 2017;10:5018.
~ 2. Capua I, Alexander DJ. Ecology and Epidemiology and human Health Implications of Avian Influenza Virus Infections in Avian Influenza and Newcastle Disease ed. Springer ,Verlag Italia 2009;1-8.
~ 3. Dimitrov KM, Abolnikb C, Afonsoa CL, et al. Updated unified phylogenetic classification system and revised nomenclature for Newcastle disease virus.Infec Genetic and Evolu 2019; 74:103917.
~ 4. Miller PJ, Koch G. Newcastle disease. Diseases of poultry. 14th. editor-in-chief, David E. Swayne; associate editors, Martine Boulianne [and 12 others], Hoboken: Wiley-Blackwell 2020: 112-29
~ 5. Awad AM, Sedeik ME, Abdelkariem AA. Isolation, molecular characterization, and pathotyping of NDV from field outbreak among broiler flocks in Egypt from 2014 to 2015. Int. J Curr Res 2015; 7: 12925-34.
~ 6. Kayali G, Kandeil A, El-Shesheny R, et al. Avian Influenza A(H5N1) Virus in Egypt. Emerg Infec Dis J 2016; 22: 379–88.
~ 7. Naeem K, Ullah A, Manvell RJ, Alexander DJ.Avian influenza A subtype H9N2 in poultry in Pakistan.Vet Rec 1999; 6:145- 560.
~ 8. Monne I, Hussein HA, Fusaro A, et al. H9N2 influenza A virus circulates in H5N1 endemically infected poultry population in Egypt. Influenza and other respiratory viruses 2013; 7: 240-3.
~ 9. Hegazy AME, Hassan AFI,Tolba HMN. Co circulation of major Avian respiratory viruses in Egypt: AIV and NDV. J of Adv Anim Vet Sci 2019; 7 :96-106.
~ 10. Kandeil A, El-Shesheny R, Maatouq AM, et al. Genetic and antigenic evolution of H9N2 avian influenza viruses circulating in Egypt between 2011 and 2013.Arch of Virol J 2013;159: 2861–76.
~ 11. El-Zoghby EF, Arafa AS, Hassan MK, et al. Isolation of H9N2 avian influenza virus from bobwhite quail (Colinus virginianus) in Egypt. Arch Virol J 2012;157: 1167-72.
~ 12. Tolba HMN, Abou Elez RMM, Elsohaby I, et al. Molecular identification of avian influenza virus subtypes H5N1 and H9N2 in birds from farms and live bird markets and in respiratory patients. Peer J 2018;1-21.
~ 13. Abdel-Moneim AS, Afifi MA, EI-Kady MA. Isolation and mutation trend analysis of influenza A virus subtype H9N2 in Egypt. Virol J 2012;9:173.
~ 14. Abdullah AS, Erfan AM, Hagag N,et al. Highly Pathogenic Avian Influenza Virus (H5N8) Clade 126.96.36.199 Infection in Migratory Birds, Egypt.Emerg Infec Dis J 2017; 23:1048-105.
~ 15. Salaheldin AH, Abd El-Hamid HS, Elbestawy AR, et al. Multiple Introductions of Influenza A (H5N8) Virus into Poultry, Egypt, 2017. Emerg Infec Dis J 2018; 24:943-6.
~ 16. Pohlmann A, Starick E, Grund C, et al. Swarm incursions of reassortants of highly pathogenic avian influenza virus strains H5N8 and H5N5, clade 188.8.131.52b, Germany, winter 2016/17. Sci Rep 2018; 8:8-15.
~ 17. Lee YJ, Kang HM, Lee EK, et al. Novel Reassortant Influenza A (H5N8) Viruses, South Korea. Emerg. Infec Dis 2014; 20:1087–9.
~ 18. Lee DH, Bertran K, Kwon JH, et al. Evolution, global spread, and pathogenicity of highly pathogenic avian influenza H5Nx clade 184.108.40.206.b. J Vet Sci 2017; 20 :269-80.
~ 19. World Organization for Animal Health (OIE). Manual on animal influenza diagnosis and virological isolation. 2015 Available at http: // www. oie. int/ fileadmin /Home /eng/Health_standards/tahm/ 2.03. 04_AI.pdf.
~ 20. Lȍndt BZ, Nunez N, Banks J, et al. Pathogenesis of highly pathogenic avian influenza A/turkey/Turkey/1/2005 H5N1 in Pekin ducks (Anas platyrhynchos) infected experimentally. Avian Pathol J 2008; 37: 619-27.
~ 21. Ben Shabat M, Meir R, Haddas R, et al. Development of a real-time TaqMan RT-PCR assay for the detection of H9N2 avian influenza viruses. J Virol Methods 2010;168 :72-7.
~ 22. Wise MG, Suarez DL, Seal BS, et al. Development of a Real-Time Reverse-Transcription PCR for Detection of Newcastle Disease Virus RNA in Clinical Samples.J of Clinic Microbiol 2004;42: 329–38.
~ 23. Shabat BM, Haddas MR, Lapin E, et al. Development of a real-time TaqMan RT-PCR assay for the detection of H9N2 avian influenza viruses. J. Virol. Methods 2010; 168:72-7.
~ 24. Thompson JD, Higgins DG, Gibson TJ. Improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. J of Nucleic Acids Res, 1994; 22:4673-80.
~ 25. Etriwati DR, Ratih D, Handharyani E, et al. Pathology and immuno histochemistry study of Newcastle disease field case in chicken in Indonesia, Veterinary World J 2017; 10:1066-1071.
~ 26. Hassan KE, Shany SAS, Ali A, et al. Prevalence of avian respiratory viruses in broiler flocks in Egypt. Poultry Science J 2016; 95:1271-80.
~ 27. Seifi S, Asasi K, Mohammadi A.Natural co-infection caused by AI H9 subtype and IB virus in broiler chicken farms. Vet. Archiv J 2010; 80:269-81.
~ 28. Kandeil A, Moatasim Y, Gomaa MR, et al. Generation of a reassortant avian influenza virus H5N2 vaccine strain capable of protecting chickens against infection with Egyptian H5N1 and H9N2 viruses. Vaccine J 2016; 34:218-24.
~ 29. Yashpal S M, Devi PP, Sagar MG. Detection of three avian Respiratory viruses by single tube multiplex reverse transcription-polymerase chain reaction assay J. Vet. Diag. Invest., 2004; 16: 244-8.
~ 30. Aly MM, Hassan MK, Arafa A. Emergence of highly pathogenic H5N1 avian influenza virus in poultry in Egypt. First record of 2006 outbreaks. J. Egypt. Vet. Med. Assoc 2006; 66: 263-76.
~ 31. Alexander DJ, Manvell RJ, Lowings JP, et al. Antigenic diversity and similarities detected in avian paramyxovirus type 1(Newcastle disease virus) isolates using monoclonal antibodies. J Avian Pathol 1997; 26:399-418.
~ 32. Tang Q, wanga J, Bao J, et al. A multiplex RT-PCR assay for detection and differentiation of avian H3, H5, and H9 subtype influenza viruses and Newcastle disease viruses. J. Virol. Methods 2012; 181:164-9.
~ 33. Saad AM, Samy A, Soliman MA, et al. Genotypic and pathogenic characterization of genotype VII Newcastle disease viruses isolated from commercial farms in Egypt and evaluation of heterologous antibody responses. Arch Virol J 2017; 162:1985–94.
~ 34. Sabra MH. Molecular characterization of Newcastle disease virus isolated from chickens in south valley provinces. M.Sc. Thesis, South Valley University, Qena, Egypt, 2013.
~ 35. Radwan M, Darwish S, El-Sabagh I, et al. Isolation and molecular characterization of Newcastle disease virus genotypes II and VIId in Egypt between 2011 and 2012 .J Virus Genes 2013; 47:311–6.
~ 36. Liu XF, Wan HQ, Ni XX, et al. Pathotypical and genotypical characterization of strains of Newcastle disease virus isolated from outbreaks in chicken and goose flocks in some regions of China during 1985-2001.J Archives of Virol 2003; 148: 1387–403.
~ 37. 37. Khan ST, Rehmani RC, Miller P, et al. Phylogenetic and pathological characterization of Newcastle disease virus isolates from Pakistan. J Clin. Microbiol 2010; 48: 1892–4.
~ 38. Arafat N, Eladl AH, Marghani BH, et al. Enhanced infection of avian influenza virus H9N2 with infectious laryngeotracheitis vaccination in chickens.J Vet.Microbiol 2018; 219: 8-16.
~ 39. Naguib MM, Arafa AS, Parvin R, et al. Insights into genetic diversity and biological propensities of potentially zoonotic avian influenza H9N2 viruses circulating in Egypt. Virol J 2017; 511:165-74.
~ 40. El-Bayoumi KM, Hassan ER, Mahgoup KM, et al. Comparative study between available used Newcastle Vaccines in market on broiler chickens. Report and Opinion J 2013; 4: 46-51.
~ 41. Ewies SS, Ali A, Tamam SM, et al. Molecular characterization of Newcastle disease virus (genotype vii) from broiler chickens in Egypt. Beni-Suef University J of Basic and Applied Sci 2017;6:232-7.
~ 42. Bouwstra R, Heutink R, Bossers A, et al. Full-Genome Sequence of Influenza A (H5N8) Virus in Poultry Linked to Sequences of Strains from Asia, the Netherlands, 2014, Emerg Infec Dis J 2015; 21: 872–4.
~ 43. Hassan KE, El-Kady MF, El-Sawah AAA, et al. Respiratory disease due to mixed viral infections in poultry flocks in Egypt between 2017 and 2018: Upsurge of highly pathogenic avian influenza virus subtype H5N8 since 2018. Transbound Emerg Dis J 2021;68:21–36
~ 44. .Kandeil AA, Kayed Y, Moatasim RJ, et al. Genetic characterization of highly pathogenic avian influenza A H5N8 viruses isolated from wild birds in Egypt. J. Gen. Virol 2017; 98: 1573–86.
~ 45. Li M, Liu H, Bi Y, et al. Highly Pathogenic Avian Influenza A(H5N8) Virus in Wild Migratory Birds, Qinghai Lake, China, Emerg Infec Dis Jl 2017; 23: 637–41.