Molecular identification of Newcastle disease virus isolated on the poultry farm of the Moscow Oblast in summer of 2022

In August 2022, a sudden death in backyard chickens was reported in the Moscow Oblast (urban district Chernogolovka, settlement Starki). As a result, within just a few days 45 chickens on this farm died or fell ill with the following signs – gray mucus discharge from nostrils and beak, coughing, gasping and rales. On day 1–3 after the onset of symptoms, the chicken died. The Newcastle disease virus, which is a representative of the Paramyxoviruses family, was isolated from the dead poultry. We determined the nucleotide sequences of fragments in F gene (encodes the fusion surface protein) and in NP gene (encodes the nucleocapsid protein). The motif of ₁₀₉SGGRRQKRFIG₁₁₉ proteolysis site, typical for the velogenic pathotype, was determined for the F gene, and a phylogenetic analysis was carried out to demonstrate that the isolate belonged to Subgenotype VII, Class II of the subfamily Avulavirinae. The Basic Local Alignment Search Tool revealed that they are most genetically related with isolates from Iran. It was found that the average death time of developing chicken embryos, infected with a minimum infectious dose, was 52 hours, which is typical for the velogenic pathotype. The virus caused 100% death in six-week-old chickens after oral infection and 100% death in all contact chickens, including those kept in cages at a distance, which proves the high level of pathogenicity and contagiousness of the recovered isolate and its ability to transmit both via fecal-oral and aerosols–borne routes. No death cases were reported in mice after intranasal infection with high doses.

1. Rima B., Balkema-Buschmann A., Dundon W. G., Duprex P., Easton A., Fouchier R., et al. ICTV virus taxonomy profile: Paramyxoviridae. J. Gen. Virol. 2019; 100 (12): 1593–1594. DOI: 10.1099/jgv.0.001328.

2. The Negative Sense Single Stranded RNA Viruses. In: Virus Taxonomy. Ed. by C. M. Fauquet, M. A. Mayo, J. Maniloff, U. Desselberger, L. A. Ball. Academic Press; 2005; 607–738. DOI: 10.1016/B978-0-12-249951-7.50014-6.

3. Alexander D. J. Newcastle disease and other avian paramyxoviruses. Rev. Sci. Tech. 2000; 19 (2): 443–462. DOI: 10.20506/rst.19.2.1231.

4. Ganar K., Das M., Sinha S., Kumar S. Newcastle disease virus: current status and our understanding. Virus Res. 2014; 184: 71–81. DOI: 10.1016/j.virusres.2014.02.016.

5. Glickman R. L., Syddall R. J., Iorio R. M., Sheehan J. P., Bratt M. A. Quantitative basic residue requirements in the cleavage-activation site of the fusion glycoprotein as a determinant of virulence for Newcastle disease virus. J. Virol. 1988; 62 (1): 354–356. DOI: 10.1128/JVI.62.1.354-356.1988.

6. Czeglédi A., Ujvári D., Somogyi E., Wehmann E., Werner O., Lomniczi B. Third genome size category of avian paramyxovirus serotype 1 (Newcastle disease virus) and evolutionary implications. Virus Res. 2006; 120 (1–2): 36–48. DOI: 10.1016/j.virusres.2005.11.009.

7. Dimitrov K. M., Abolnik C., Afonso C. L., Albina E., Bahl J., Berg M., et al. Updated unified phylogenetic classification system and revised nomenclature for Newcastle disease virus. Infect. Genet. Evol. 2019; 74:103917. DOI: 10.1016/j.meegid.2019.103917.

8. Beard C. W., Hanson R. P. Newcastle disease. In: Diseases of Poultry. Ed. by M. S. Hofstad, H. J. Barnes, B. W. Calnek, W. M. Reid, H. W. Yoder, et al. 8th ed. Ames: Iowa State University Press; 1984; 452–470.

9. Miller P. J., Haddas R., Simanov L., Lublin A., Rehmani S. F., Wajid A., et al. Identification of new sub-genotypes of virulent Newcastle disease virus with potential panzootic features. Infect. Genet. Evol. 2015; 29: 216–229. DOI: 10.1016/j.meegid.2014.10.032.

10. Karamendin K., Kydyrmanov A. Cormorants as a potentially important reservoir and carrier of Newcastle disease virus on the Asian continent. Front. Vet. Sci. 2021; 8:648091. DOI: 10.3389/fvets.2021.648091.

11. Shengqing Y., Kishida N., Ito H., Kida H., Otsuki K., Kawaoka Y., Ito T. Generation of velogenic Newcastle disease viruses from a nonpathogenic waterfowl isolate by passaging in chickens. Virology. 2002; 301 (2): 206–211. DOI: 10.1006/viro.2002.1539.

12. Newcastle disease (infection with Newcastle disease virus). In: WOAH. Manual of Diagnostic Tests and Vaccines for Terrestrial Animals. 2021; Chapter 3.3.14. Available at: https://www.woah.org/fileadmin/Home/eng/Health_standards/tahm/3.03.14_NEWCASTLE_DIS.pdf.

13. Staden R. The Staden sequence analysis package. Mol. Biotechnol. 1996; 5 (3): 233–241. DOI: 10.1007/BF02900361.

14. Kumar S., Stecher G., Li M., Knyaz C., Tamura K. MEGA X: molecular evolutionary genetics analysis across computing platforms. Mol. Biol. Evol. 2018; 35 (6): 1547–1549. DOI: 10.1093/molbev/msy096.

15. Drummond A. J., Rambaut A., Shapiro B., Pybus O. G. Bayesian coalescent inference of past population dynamics from molecular sequences. Mol. Biol. Evol. 2005; 22 (5): 1185–1192. DOI: 10.1093/molbev/msi103.

16. Tan S. W., Ideris A., Omar A. R., Yusoff K., Hair-Bejo M. Detection and differentiation of velogenic and lentogenic Newcastle disease viruses using SYBR Green I real-time PCR with nucleocapsid gene-specific primers. J. Virol. Methods. 2009; 160 (1–2): 149–156. DOI:10.1016/j.jviromet.2009.05.006.

17. Eid A. A. M., Hussein A., Hassanin O., Elbakrey R. M., Daines R., Sadeyen J. R., et al. Newcastle disease genotype VII prevalence in poultry and wild birds in Egypt. Viruses. 2022; 14 (10):2244. DOI: 10.3390/v14102244.

18. Xue C., Xu X., Yin R., Qian J., Sun Y., Wang C., et al. Identification and pathotypical analysis of a novel VIk sub-genotype Newcastle disease virus obtained from pigeon in China. Virus Res. 2017; 238: 1–7. DOI: 10.1016/j.virusres.2017.05.011.

19. Perozo F., Marcano R., Afonso C. L. Biological and phylogenetic characterization of a genotype VII Newcastle disease virus from Venezuela: efficacy of field vaccination. J. Clin. Microbiol. 2012; 50 (4): 1204–1208. DOI: 10.1128/JCM.06506-11.

20. Pandarangga P., McAllister M. M., Peaston A. E., Ngai Y. T., Cahyono M. I., Hemmatzadeh F. Performance comparison of homologous and heterologous Newcastle disease virus in vaccines and antibody tests. Res. Vet. Sci. 2022; 149: 82–89. DOI: 10.1016/j.rvsc.2022.06.014.

21. Miller P. J., Decanini E. L., Afonso C. L. Newcastle disease: evolution of genotypes and the related diagnostic challenges. Infect. Genet. Evol. 2010; 10 (1): 26–35. DOI: 10.1016/j.meegid.2009.09.012.

22. Gogoi P., Ganar K., Kumar S. Avian paramyxovirus: a brief review. Transbound. Emerg. Dis. 2017; 64 (1): 53–67. DOI: 10.1111/tbed.12355.

23. Volkova M. A., Chvala Ir. A., Yaroslavtseva P. S., Sosipatorova V. Yu., Chvala I. A. Serological monitoring of Newcastle disease in Russia 2017. Veterinary Science Today. 2018; (4): 26–30. DOI: 10.29326/2304-196X-2018-4-27-26-30.

24. Volkova M. A., Chvala Ir. A., Osipova O. S., Kulagina M. A., Andreychuk D. B., Chvala I. A. Serological monitoring of avian influenza and Newcastle disease in the Russian Federation in 2019. Veterinary Science Today. 2020; (2): 76–82. DOI: 10.29326/2304-196X-2020-2-33-76-82.

25. Kulagina M. A., Volkova M. A., Chvala Ir. A., Osipova O. S., Yaroslavtseva P. S., Andreychuk D. B., Chvala I. A. Serological monitoring of avian influenza and Newcastle disease in the Russian Federation in 2020. Veterinary Science Today. 2022; 11 (2): 142–148. DOI: 10.29326/2304-196X-2022-11-2-142-148.

26. Frolov S. V., Moroz N. V., Chvala I. A., Irza V. N. Effectiveness of vaccines produced by the Federal State-Financed Institution “ARRIAH” against topical genotype VII Newcastle disease viruses. Veterinary Science Today. 2021; (1): 44–51. DOI: 10.29326/2304-196X-2021-1-36-44-51.

27. Instructions on control measures against Newcastle disease (pseudo-poultry plague): approved by the Chief Veterinary Department of the USSR MoA on 09 June 1976 (with amendments and additions dated back to August 28, 1978). Available at: http://vetorel.ru/website/img/docs/newak.pdf.

28.