NDV subgenotype VII(L) is currently circulating in commercial broiler farms of Iran, 2017–2018

  • Aidin Molouki
  • Mohammad Hossein Fallah Mehrabadi
  • Mohsen BashashatiEmail author
  • Mohsen Mahmoudzadeh Akhijahani
  • Swee Hua Erin Lim
  • Saeed Amir Hajloo
Regular Articles



Based on our previous work, it was discovered that some Newcastle disease virus (NDV) isolates from backyard poultry between 2011 and 2013 in Iran formed a new separate cluster when phylogenetic analysis based on the complete F gene sequence was carried out. The novel cluster was designated subgenotype VII(L) and published.


In the current study, for further validation, we initiated a comprehensive epidemiological study to identify the dominant NDV genotype(s) circulating within the country. Collection of samples was executed between October 2017 and February 2018 from 108 commercial broiler farms which reported clinical signs of respiratory disease in their broilers.


We report that 38 of the farms (> 35%) tested positive for NDV. The complete F gene sequences of seven of the isolates are shown as representative sequences in this study. According to the phylogenetic tree constructed, the recent broiler farm isolates clustered into the newly designated cluster VII(L) together with the older Iranian backyard poultry isolates in our previous work. All the sequences shared the same virulence-associated F cleavage site of 112RRQKR↓F117.


Our phylogenetic analysis suggested that the NDV subgenotype VII(L) may have been derived from subgenotype VIId, and contrary to popular belief, subgenotype VIId may not be the dominant subgenotype in Iran. Tracking of the subgenotype on BLAST suggested that the NDV subgenotype VII(L), although previously unidentified, may have been circulating in this region as an endemic virus for at least a decade. Other NDV genotypes, however, have also been reported in Iran in recent years. Hence, ongoing study is aimed at determining the exact dominant NDV genotypes and subgenotypes in the country. This will be crucial in effective mitigation of outbreaks in Iranian broiler farms.


NDV Subgenotype VII(L) Outbreak Poultry diseases Phylogenetic study 



Newcastle disease virus


Iranian Veterinary Organization


hemagglutination assay


Author contributions

SAH and MHFM collected the samples from broiler farms. MMA, AM, MHFM, and MB inoculated the samples in SPF eggs and harvested. MMA, AM, and MHFM performed the rapid hemagglutination test. MMA performed HA, HI, and MDT. AM and MB extracted RNA, synthesized cDNA, and ran PCR. AM designed the primers. AM cloned the PCR fragments, transformed, extracted plasmids, and prepared for sequencing. AM assembled the sequencing results and submitted to GenBank. MB did the phylogenetic analysis. AM wrote the first draft of the manuscript. SHEL, MB, and MHFM edited the manuscript.

Funding information

AM, MHFM, and MB were supported by Razi Vaccine and Serum Research Institute grant number 1218181296006960282.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Research involving animals

All procedures involving human participant was in accordance with ethical standards.

Informed consent

Informed consent was obtained from all individual participants included in the study.


  1. Creelan, J.L., Graham, D.A., and McCullough, S.J., 2002. Detection and differentiation of pathogenicity of avian paramyxovirus serotype 1 from field cases using one-step reverse transcriptase-polymerase chain reaction. Avian Pathol 31, 493–499CrossRefGoogle Scholar
  2. de Leeuw, O.S., Koch, G., Hartog, L., Ravenshorst, N., and Peeters, B.P., 2005. Virulence of Newcastle disease virus is determined by the cleavage site of the fusion protein and by both the stem region and globular head of the haemagglutinin-neuraminidase protein. J Gen Virol 86, 1759–1769CrossRefGoogle Scholar
  3. Diel, D.G., da Silva, L.H.A., Liu, H., Wang, Z., Miller, P.J., and Afonso, C.L., 2012. Genetic diversity of avian paramyxovirus type 1: proposal for a unified nomenclature and classification system of Newcastle disease virus genotypes. Infect Genet Evol 12, 1770–1779CrossRefGoogle Scholar
  4. Dimitrov, K.M., Afonso, C.L., Yu, Q., and Miller, P.J., 2016. Newcastle disease vaccines-a solved problem or a continuous challenge? Vet Microbiol 206, 126–136CrossRefGoogle Scholar
  5. Dortmans, J., Koch, G., Rottier, P., and Peeters, B., 2011. Virulence of newcastle disease virus: what is known so far? Vet Res 42, 122CrossRefGoogle Scholar
  6. Esmaelizad, M., Mayahi, V., Pashaei, M., and Goudarzi, H., 2017. Identification of novel Newcastle disease virus sub-genotype VII-(j) based on the fusion protein. Arch Virol 162, 971–978CrossRefGoogle Scholar
  7. Estevez, C., King, D.J., Luo, M., and Yu, Q., 2011. A single amino acid substitution in the haemagglutinin-neuraminidase protein of Newcastle disease virus results in increased fusion promotion and decreased neuraminidase activities without changes in virus pathotype. J Gen Virol 92, 544–551CrossRefGoogle Scholar
  8. Ghalyanchilangeroudi, A., Hosseini, H., Jabbarifakhr, M., Fallah Mehrabadi, M.H., Najafi, H., Ghafouri, S.A., Mousavi, F.S., Ziafati, Z., and Modiri, A., 2018. Emergence of a virulent genotype VIIi of Newcastle disease virus in Iran. Avian Pathol 47, 509–519CrossRefGoogle Scholar
  9. Haji-Abdolvahab, H., Ghalyanchilangeroudi, A., Bahonar, A., Ghafouri, S.A., Vasfi Marandi, M., Mehrabadi, M.H.F., and Tehrani, F., 2018. Prevalence of avian influenza, Newcastle disease, and infectious bronchitis viruses in broiler flocks infected with multifactorial respiratory diseases in Iran, 2015-2016. Trop Anim Health Prod.
  10. Kumar, S., Stecher, G., and Tamura, K., 2016. MEGA7: molecular evolutionary genetics analysis version 7.0 for bigger datasets. Mol Biol Evol 33, 1870–1874CrossRefGoogle Scholar
  11. Mehrabadi, M.H.F., Bahonar, A., Mirzaei, K., Molouki, A., Ghalyanchilangeroudi, A., Ghafouri, S.A., Tehrani, F., and Lim, S.H.E., 2017. Prevalence of avian influenza (H9N2) in commercial quail, partridge, and turkey farms in Iran, 2014-2015. Trop Anim Health Prod 50, 677–682CrossRefGoogle Scholar
  12. Miller, P.J., King, D.J., Afonso, C.L., and Suarez, D.L., 2007. Antigenic differences among Newcastle disease virus strains of different genotypes used in vaccine formulation affect viral shedding after a virulent challenge. Vaccine 25, 7238–7246CrossRefGoogle Scholar
  13. Miller, P.J., Haddas, R., Simanov, L., Lublin, A., Rehmani, S.F., Wajid, A., Bibi, T., Khan, T.A., Yaqub, T., Setiyaningsih, S., and Afonso, C.L., 2015. Identification of new sub-genotypes of virulent Newcastle disease virus with potential panzootic features. Infect Genet Evol 29, 216–229CrossRefGoogle Scholar
  14. Molouki, A., and Peeters, B., 2017. Rescue of recombinant Newcastle disease virus: current cloning strategies and RNA polymerase provision systems. Arch Virol 162, 1–12CrossRefGoogle Scholar
  15. OIE, 2004. Newcastle disease. Manual of diagnostic tests and vaccines for terrestrial animals, 2004, (World Organisation for Animal Health, Paris, France), 270–282Google Scholar
  16. OIE, 2012. Newcastle disease. Manual of diagnostic tests and vaccines for terrestrial animals, 2012, (World Organisation for Animal Health, Paris, France), 555–573Google Scholar
  17. Peeters, B.P., de Leeuw, O.S., Koch, G., and Gielkens, A.L., 1999. Rescue of Newcastle disease virus from cloned cDNA: evidence that cleavability of the fusion protein is a major determinant for virulence. J Virol 73, 5001–5009Google Scholar
  18. Sabouri, F., Vasfi Marandi, M., and Bashashati, M., 2018. Characterization of a novel VIIl sub-genotype of Newcastle disease virus circulating in Iran. Avian Pathol 47, 90–99CrossRefGoogle Scholar
  19. Saitou, N., and Nei, M., 1987. The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4, 406–425Google Scholar
  20. Sambrook, J., Russell, D.W., and Sambrook, J., 2006. The condensed protocols from Molecular cloning : a laboratory manual, (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.)Google Scholar
  21. Sohrab, V., 1973. Newcastle disease in Iran. Bull Off Int Epiz 79, 565–569Google Scholar
  22. Tamura, K., Nei, M., and Kumar, S., 2004. Prospects for inferring very large phylogenies by using the neighbor-joining method. Proc Natl Acad Sci USA 101, 11030–11035CrossRefGoogle Scholar
  23. WAHIS, 2018. 2018,Google Scholar

Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.Department of Avian Disease Research and Diagnostic, Razi Vaccine and Serum Research InstituteAgricultural Research Education and Extension Organization (AREEO)KarajIran
  2. 2.Health Sciences Division, Abu Dhabi Women’s CollegeHigher Colleges of TechnologyAbu DhabiUnited Arab Emirates
  3. 3.Centre for Research Excellence and School of Data SciencePerdana UniversitySerdangMalaysia
  4. 4.Department of Health and Management of Poultry DiseaseIranian Veterinary Organization (IVO)TehranIran

Personalised recommendations