Detection of Virulent Genotype VII Newcastle Disease Virus in Lebanese Poultry Using Partial F Gene Sequences Reveals Regional and Global Genetic Relatedness — Oak Academic Publishing
Research ArticleOpen AccessGoogle Scholar indexed
Detection of Virulent Genotype VII Newcastle Disease Virus in Lebanese Poultry Using Partial F Gene Sequences Reveals Regional and Global Genetic Relatedness
Department of Agriculture, Faculty of Agricultural and Food Sciences, American University of Beirut, Beirut, Lebanon
,
Department of Agriculture, Faculty of Agricultural and Food Sciences, American University of Beirut, Beirut, Lebanon
,
Department of Agriculture, Faculty of Agricultural and Food Sciences, American University of Beirut, Beirut, Lebanon
1 Department of Agriculture, Faculty of Agricultural and Food Sciences, American University of Beirut, Beirut, Lebanon
2 Department of Agriculture, Faculty of Agricultural and Food Sciences, American University of Beirut, Beirut, Lebanon
3 Department of Agriculture, Faculty of Agricultural and Food Sciences, American University of Beirut, Beirut, Lebanon
Newcastle disease virus (NDV) continues to threaten poultry industries worldwide, with genotype VII strains currently dominating outbreaks across the Middle East. In Lebanon, data on circulating NDV lineages remain scarce. This study molecularly characterizes three NDV isolates obtained in Spring 2025 from commercial and backyard poultry farms in Lebanon, where birds exhibited respiratory and/or neurological signs. Virus propagation was conducted in embryonated chicken eggs, and successful replication was confirmed by hemagglutination activity. Reverse transcription PCR targeting a 254 bp fragment of the fusion (F) gene consistently yielded the expected amplicons from all samples. Sequencing revealed the virulent polybasic cleavage site motif ^112RRQKR^117 in all isolates. Phylogenetic analysis placed the Lebanese viruses within genotype VII cluster, showing 100% nucleotide identity with an Indonesian reference strain and close clustering with isolates from neighboring Middle Eastern and North African countries, as well as South and Southeast Asia. Despite different vaccination profiles, all isolates exhibited identical partial F gene sequence. These findings highlight the dominance and persistence of genotype VII NDV in the region, its genetic relatedness to globally circulating strains, and the potential challenges for vaccine efficacy. Continuous molecular surveillance and evaluation of vaccine performance are essential for improving NDV control strategies in Lebanon and the region.
KeywordsNewcastle Disease Virus (NDV)Genotype VIIFusion Protein Cleavage SitePhylogenetic AnalysisLebanon
Afonso, C.L. (2021) Virulence during Newcastle Disease Viruses Cross Species Adaptation. Viruses , 13, Article 110. https://doi.org/10.3390/v13010110
Swayne, D.E. (2020) Diseases of Poultry. 14th Edition, Wiley-Blackwell.
Wang, Y., Yu, W., Huo, N., Wang, W., Guo, Y., Wei, Q., et al. (2017) Comprehensive Analysis of Amino Acid Sequence Diversity at the F Protein Cleavage Site of Newcastle Disease Virus in Fusogenic Activity. PLOS ONE , 12, e0183923. https://doi.org/10.1371/journal.pone.0183923
Eid, A.A.M., Hussein, A., Hassanin, O., Elbakrey, R.M., Daines, R., Sadeyen, J., et al. (2022) Newcastle Disease Genotype VII Prevalence in Poultry and Wild Birds in Egypt. Viruses , 14, Article 2244. https://doi.org/10.3390/v14102244
Orabi, A., Hussein, A., Saleh, A.A., El-Magd, M.A. and Munir, M. (2017) Evolutionary Insights into the Fusion Protein of Newcastle Disease Virus Isolated from Vaccinated Chickens in 2016 in Egypt. Archives of Virology , 162, 3069-3079. https://doi.org/10.1007/s00705-017-3483-1
Ghalyanchilangeroudi, A., Hosseini, H., Jabbarifakhr, M., Fallah Mehrabadi, M.H., Najafi, H., Ghafouri, S.A., et al. (2018) Emergence of a Virulent Genotype VIII of Newcastle Disease Virus in Iran. Avian Pathology , 47, 509-519. https://doi.org/10.1080/03079457.2018.1495313
Miller, P.J., Haddas, R., Simanov, L., Lublin, A., Rehmani, S.F., Wajid, A., et al. (2015) Identification of New Sub-Genotypes of Virulent Newcastle Disease Virus with Potential Panzootic Features. Infection , Genetics and Evolution , 29, 216-229. https://doi.org/10.1016/j.meegid.2014.10.032
Al‐Shammari, A.M., Hamad, M.A., AL‐Mudhafar, M.A., Raad, K. and Ahmed, A. (2020) Clinical, Molecular and Cytopathological Characterization of a Newcastle Disease Virus from an Outbreak in Baghdad, Iraq. Veterinary Medicine and Science , 6, 477-484. https://doi.org/10.1002/vms3.262
Mayahi, V. and Esmaelizad, M. (2017) Molecular Evolution and Epidemiological Links Study of Newcastle Disease Virus Isolates from 1995 to 2016 in Iran. Archives of Virology , 162, 3727-3743. https://doi.org/10.1007/s00705-017-3536-5
Naguib, M.M., Höper, D., Elkady, M.F., Afifi, M.A., Erfan, A., Abozeid, H.H., et al. (2021) Comparison of Genomic and Antigenic Properties of Newcastle Disease Virus Genotypes II, XXI and VII from Egypt Do Not Point to Antigenic Drift as Selection Marker. Transboundary and Emerging Diseases , 69, 849-863. https://doi.org/10.1111/tbed.14121
Yang, H., Zhao, J., Xue, J., Yang, Y. and Zhang, G. (2017) Antigenic Variation of Lasota and Genotype VII Newcastle Disease Virus (NDV) and Their Efficacy against Challenge with Velogenic NDV. Vaccine , 35, 27-32. https://doi.org/10.1016/j.vaccine.2016.11.048
Roohani, K., Tan, S.W., Yeap, S.K., Ideris, A., Bejo, M.H. and Omar, A.R. (2015) Characterisation of Genotype VII Newcastle Disease Virus (NDV) Isolated from NDV Vaccinated Chickens, and the Efficacy of Lasota and Recombinant Genotype VII Vaccines against Challenge with Velogenic NDV. Journal of Veterinary Science , 16, 447-457. https://doi.org/10.4142/jvs.2015.16.4.447
Abdelsabour, M.A., Helal, A.M., Nagar, E.M.S.E., El-fatah, W.A., Abodalal, S.E.S.A., Madbouly, Y.M., Arafa, A.A., Ibrahim, H.M. and Hussein, A. (2024) Efficacy of a Locally Prepared Live Clone Vaccine against Newcastle Disease Virus Genotype IV and Genotype VIID in Egypt: Clone 30 Vaccine. Journal of Advanced Veterinary Re-search , 14, 639-643. https://advetresearch.com/index.php/AVR/article/view/1746
Barbour, E. (2017) Protection by Classical and Vectored Vaccines against Endemic Velogenic Newcastle Disease in Broiler Farms and Evaluation of a Developed Autogenous Vaccine against Predominant Genotype VI. International Journal of Vaccines & Vaccination , 4, Article ID: 00075. https://doi.org/10.15406/ijvv.2017.04.00075
Dufour-Zavala, L. and American Association of Avian Pathologists (2008) A Laboratory Manual for the Isolation, Identification and Characterization of Avian Pathogens. 5th Edition, American Association of Avian Pathologists.
Kant, A., Koch, G., Van Roozelaar, D.J., Balk, F. and Huurne, A.T. (1997) Differentiation of Virulent and Non-Virulent Strains of Newcastle Disease Virus within 24 Hours by Polymerase Chain Reaction. Avian Pathology , 26, 837-849. https://doi.org/10.1080/03079459708419257
Patel, S.S., Chauhan, H.C., Kumar Sharma, K., Patel, A.C., Bulbule, N.R., Raval, S.H., et al. (2024) Genetic Evolution of Newcastle Disease Virus Sub-Genotype VII.2 Isolates, Diagnosed from Vaccinated Poultry Farms of Gujarat, India. Gene , 930, Article ID: 148859. https://doi.org/10.1016/j.gene.2024.148859
Dharmayanti, N.I., Nurjanah, D., Nuradji, H., Suyatno, T. and Indriani, R. (2024) Newcastle Disease Virus: The Past and Current Situation in Indonesia. Journal of Veterinary Science , 25, e3. https://doi.org/10.4142/jvs.23022
He, Y., Taylor, T.L., Dimitrov, K.M., Butt, S.L., Stanton, J.B., Goraichuk, I.V., et al. (2018) Whole-Genome Sequencing of Genotype VI Newcastle Disease Viruses from Formalin-Fixed Paraffin-Embedded Tissues from Wild Pigeons Reveals Continuous Evolution and Previously Unrecognized Genetic Diversity in the U.S. Virology Journal , 15, Article No. 9. https://doi.org/10.1186/s12985-017-0914-2
Radwan, M.M., Darwish, S.F., El-Sabagh, I.M., El-Sanousi, A.A. and Shalaby, M.A. (2013) Isolation and Molecular Characterization of Newcastle Disease Virus Genotypes II and VIID in Egypt between 2011 and 2012. Virus Genes , 47, 311-316. https://doi.org/10.1007/s11262-013-0950-y
Bello, M.B., Yusoff, K., Ideris, A., Hair-Bejo, M., Peeters, B.P.H. and Omar, A.R. (2018) Diagnostic and Vaccination Approaches for Newcastle Disease Virus in Poultry: The Current and Emerging Perspectives. BioMed Research International , 2018, Article ID: 7278459. https://doi.org/10.1155/2018/7278459
Dimitrov, K.M., Lee, D., Williams-Coplin, D., Olivier, T.L., Miller, P.J. and Afonso, C.L. (2016) Newcastle Disease Viruses Causing Recent Outbreaks Worldwide Show Unexpectedly High Genetic Similarity to Historical Virulent Isolates from the 1940s. Journal of Clinical Microbiology , 54, 1228-1235. https://doi.org/10.1128/jcm.03044-15
Sultan, H.A., Elfeil, W.K., Nour, A.A., Tantawy, L., Kamel, E.G., Eed, E.M., et al. (2021) Efficacy of the Newcastle Disease Virus Genotype VII.1.1-Matched Vaccines in Commercial Broilers. Vaccines , 10, Article 29. https://doi.org/10.3390/vaccines10010029
Mahamud, S.N.A., Bello, M.B., Ideris, A. and Omar, A.R. (2022) Efficacy of Genotype-Matched Newcastle Disease Virus Vaccine Formulated in Carboxymethyl Sago Starch Acid Hydrogel in Chickens Vaccinated via Different Routes. Journal of Veterinary Science , 23, e25. https://doi.org/10.4142/jvs.21242
Bello, M.B., Mahamud, S.N.A., Yusoff, K., Ideris, A., Hair-Bejo, M., Peeters, B.P.H., et al. (2020) Development of an Effective and Stable Genotype-Matched Live Attenuated Newcastle Disease Virus Vaccine Based on a Novel Naturally Recombinant Malaysian Isolate Using Reverse Genetics. Vaccines , 8, Article 270. https://doi.org/10.3390/vaccines8020270
Otiang, E., Thumbi, S.M., Campbell, Z.A., Njagi, L.W., Nyaga, P.N. and Palmer, G.H. (2021) Impact of Routine Newcastle Disease Vaccination on Chicken Flock Size in Smallholder Farms in Western Kenya. PLOS ONE , 16, e0248596. https://doi.org/10.1371/journal.pone.0248596
Liu, H., de Almeida, R.S., Gil, P., Majó, N., Nofrarías, M., Briand, F., et al. (2018) Can Genotype Mismatch Really Affect the Level of Protection Conferred by Newcastle Disease Vaccines against Heterologous Virulent Strains? Vaccine , 36, 3917-3925. https://doi.org/10.1016/j.vaccine.2018.05.074
Kapczynski, D.R., Afonso, C.L. and Miller, P.J. (2013) Immune Responses of Poultry to Newcastle Disease Virus. Developmental & Comparative Immunology , 41, 447-453. https://doi.org/10.1016/j.dci.2013.04.012