Biological characterization of wild Newcastle disease virus strains Isolated in Russia between 2018 and 2021

Newcastle disease virus (NDV) continues to be a significant pathogen of avian species, with wild migratory birds playing a pivotal role in its circulation and interregional dissemination. This study aimed to investigate the biological characteristics of NDV strains isolated from wild birds in Russia, focusing on their replication efficiency and cytotoxic activity against mammalian tumor cell lines. Between 2018 and 2021, 92 samples were collected from wild waterfowl, and viruses were subsequently isolated in embryonated chicken eggs. Viral titers were determined using hemagglutination assays and VeroE6 cell cultures. Cytotoxicity of 19 NDV strains was evaluated on HeLa, SW13, RD, SKBR3, HEp‐2, and A549 cell lines. Cell viability was assessed using the MTT assay. The NDV strains exhibited substantial variability in both replication and cytotoxic activity. The most pronounced cytopathic effects were observed in SW13 and RD cells, whereas SKBR3, HeLa, and HEp‐2 cells displayed lower sensitivity. Strains with both high and low cytotoxic potential were identified, reflecting the heterogeneity of the isolates. These findings highlight the biological diversity of natural NDV isolates and their differential interactions with mammalian cells, which is relevant for assessing epizootic risk and exploring the virus’s potential as an oncolytic agent.

1. Malik Y.S., Arun Prince Milton A., Ghatak S., Ghosh S. Newcastle Disease and Other Avian Paramyxoviruses. Role of Birds in Transmitting Zoonotic Pathogens. Livestock Diseases and Management. Springer, Singapore, 2021, pp. 79–91. https://doi.org/10.1007/978‐981‐16‐4554‐9_6

2. Ariyama N., Tapia R., Godoy C., Agüero B., Valdés V., Berrios F., García Borboroglu P., Pütz K., Alegria R., Barriga G.P., Medina R., Neira V. Avian orthoavulavirus 1 (Newcastle Disease virus) antibodies in five penguin species, Antarctic peninsula and Southern Patagonia. Transboundary and Emerging Diseases, 2021, vol. 68, no. 6, pp. 3096–3102. https://doi.org/10.1111/tbed.14037

3. Samad A., Hamza M., Muazzam A., Ahmer A., Tariq S., Shahid M.J., Akram W., Kaleem M.Z., Javaid A. Newcastle disease in poultry, its diagnosis, prevention and control strategies. International Journal of Medical, Pharmaceutical and Drug Research, 2022, vol. 6, iss. 2, pp. 25‒28. https://doi.org/10.22161/IJMPD.6.2.3

4. Murashkina T., Sharshov K., Gadzhiev A., Petherbridge G., Derko A., Sobolev I., Dubovitskiy N., Loginova A., Kurskaya O., Kasianov N., Kabilov M., Mine J., Uchida Y., Tsunekuni R., Saito T., Alekseev A., Shestopalov A. Avian Influenza Virus and Avian Paramyxoviruses in Wild Waterfowl of the Western Coast of the Caspian Sea (2017‐2020). Viruses, 2024, vol. 16, no. 4, pp. 598. doi: 10.3390/v16040598

5. Afonso C.L. Virulence during Newcastle Disease Viruses Cross Species Adaptation. Viruses, 2021, vol. 13, no. 1, pp. 110. doi: 10.3390/v13010110

6. Kanwal A., Rashid M., Shakir U., Rizwan M., Akram M.S. Role of wild birds in spreading potential zoonotic diseases in poultry. Zoonosis, 2023, vol. 1, pp. 530–540.

7. Derko A., Dubovitskiy N., Prokudin A., Mine J., Tsunekuni R., Uchida Y., Saito T., Kasianov N., Loginova A., Sobolev I., Kumar S., Shestopalov A., Sharshov K. Detection of a Novel Gull‐like Clade of Newcastle Disease Virus and H3N8 Avian Influenza Virus in the Arctic Region of Russia (Taimyr Peninsula). Viruses, 2025, vol. 17, no. 7, pp. 955. doi: 10.3390/v17070955

8. Glushchenko A.V., Yurchenko K.S., Yurlov A.K., Yushkov Y.G., Shchelkanov M.Y., Shestopalov A.M. The role of wild birds in preservation and prevalence of avian paramyxovirus serotype 1 (Newcastle disease viruses) in Siberia and the Far East, Russia. South of Russia: ecology, development, 2016, vol. 11, no. 2, pp. 50–58. (In Russian) https://doi.org/10.18470/1992‐1098‐2016‐2‐50‐58

9. Zhou J., Duan Y., Liu M., Liu J., Hu Z., Duan Z. Recent advancements in the diverse roles of polymerase‐associated proteins in the replication and pathogenesis of Newcastle disease virus. Veterinary Research, 2025, vol. 56, no. 1, p. 8.

10. Guseva N.A., Kolosov S.N., Zinyakov N.G., Andriyasov A.V., Yin R., Scherbakova L.O., Ovchinnikova E.V., Nikonova Z.B., Andreychuk D.B., Sprygin A.V., Chvala I.A., Moroz N.V. Analysis of avian orthoavulavirus 1 detected in the Russian Federation between 2017 and 2021. Vaccines, 2023, vol. 11, no. 6, p. 1032.

11. Perlas A., Bertran K., Abad F.X., Borrego C.M., Nofrarías M., Valle R., Pailler‐García L., Ramis A., Cortey M., Acuña V., Majó N. Persistence of low pathogenic avian influenza virus in artificial streams mimicking natural conditions of waterfowl habitats in the Mediterranean climate. Science of the Total Environment, 2023, vol. 863, article id: 160902.

12. Karamendin K., Kydyrmanov A. Cormorants as a potentially important reservoir and carrier of Newcastle disease virus on the Asian continent. Frontiers in Veterinary Science, 2021, vol. 8, article id: 648091.

13. Amoia C.F., Hakizimana J.N., Chengula A.A., Munir M., Misinzo G., Weger‐Lucarelli J. Genomic diversity and geographic distribution of Newcastle disease virus genotypes in Africa: implications for diagnosis, vaccination, and regional collaboration. Viruses, 2024, vol. 16, no. 5, p. 795.

14. Nooruzzaman, M. et al. Genetic and biological characterization of Newcastle disease viruses circulating in Bangladesh during 2010–2017: further genetic diversification of class II genotype XIII in Southcentral Asia. Journal of General Virology, 2021, vol. 102, no. 3, article id: 001554.

15. Zhang D., Ding Z., Xu X. Pathologic mechanisms of the Newcastle disease virus. Viruses, 2023, vol. 15, no. 4, p. 864.

16. Maciel C. World Organisation for Animal Health. In: Elgar Concise Encyclopedia of Animal Law, 2025, p. 446.

17. Karkavin Y.I., Adamenko L.S., Yurchenko K.S., Glushchenko A.V. Antitumor effect of the Newcastle disease virus strain NDV/Altai/pigeon/777/2010 on a model of solid Lewis carcinoma. South of Russia: ecology, development, 2024, vol. 19, no. 3, pp. 44–54. (In Russian) doi: 10.18470/1992‐1098‐2024‐3‐4

18. Yurchenko K.S., Adamenko L.S., Glushchenko A.V., Shestopalov A.M. Oncolytic virotherapy with mesogenic Newcastle disease virus for Lewis lung carcinoma. Cell and Tissue Biology, 2025, vol. 19, no. 4, pp. 347–357.

19. Huang F., Dai C., Zhang Y., Zhao Y., Wang Y., Ru G. Development of molecular mechanisms and their application on oncolytic Newcastle disease virus in cancer therapy. Frontiers in Molecular Biosciences, 2022, vol. 9, article id: 889403.

20. Lu X., Zhan T., Zhou Q., Yang W., Liu K., Chen Y., Gao R., Hu J., Gu M., Hu S., Jiao X.A., Wang X., Liu X., Liu X. The haemagglutinin‐neuraminidase protein of velogenic Newcastle disease virus enhances viral infection through NF‐κB‐mediated programmed cell death. Veterinary Research, 2024, vol. 55, no. 1, p. 58.

21. Bian J., Wang K., Kong X., Liu H., Chen F., Hu M., Zhang X., Jiao X., Ge B., Wu Y., Meng S. Caspaseand p38‐MAPKdependent induction of apoptosis in A549 lung cancer cells by Newcastle disease virus. Archives of virology, 2011, vol. 156, no. 8, pp. 1335–1344.

22. Chandrasekaran A.P., Poondla N., Ko N.R., Oh S.J., Ramakrishna S. YM155 sensitizes HeLa cells to TRAILmediated apoptosis via cFLIP and survivin downregulation. Oncology Letters, 2020, vol. 20, no. 4, p. 72.

23. Miller P.J., Haddas R., Simanov L., Lublin A., Rehmani S.F., Wajid A., Bibi T., Khan T.A., Yaqub T., Setiyaningsih S., Afonso C.L. Identification of new sub‐genotypes of virulent Newcastle disease virus with potential panzootic features. Infection, Genetics and Evolution, 2015, vol. 29, pp. 216–229.