Adaptation of velogenic Newcastle disease virus in Vero cells: velogenicity of virus unaltered after adaptation


Published: Nov 22, 2017
Keywords:
Newcastle disease virus adaptation Vero cells
P. V. RAVINDRA
B. RATTA
U. CHATURVEDI
S. K. PALIA
P. K SUBUDHI
A. K. TIWARI
Abstract

In the present experiment, velogenic Newcastle disease virus (NDV) was adapted to Vero cells without the addition of exogenous trypsin. The cytopathic effect (CPE) in NDV-infected Vero cells was observed from passage 3 (P 3) onwards with concomitant increase in the haemagglutination (HA) and virus titres. The CPE was characterized by rounding of cells, their detachment from the surface and syncytia formation. The Vero-adapted NDV was confirmed by reverse transcription-polymerase chain reaction (RT-PCR), which detected the amplification of haemagglutinin neuraminidase (HN) gene from virus-infected cells. The mean death time (MDT) of 58 hours upon inoculation to embryonated chicken eggs and the intravenous pathogenicity index (IVPI) of 2.12 following inoculation to SPF chicken suggested that Vero adapted virus retained the velogenicity even after ten passages. Furthermore, the nervous signs and haemorrhages in the pro-ventriculus of birds that died at day 5 post inoculation with PIO virus confirmed that the Vero-adapted NDV was a neurotropic viscerotropic velogenic strain.

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References
Ahamed T, Hossain Κ M, Billah M M, Islam Κ M D, Ahasan M M, Islam M E (2004) Adaptation of Newcastle disease virus (NDV) on Vero cell line. Int. J. Poultry Sci., 3:153-156.
Alexander D J (1997) Newcastle disease and other Paramyxoviridae infections, In: Diseases of poultry, 10th ed. B.W. Calnek, H. J. Barnes, C.W. Beard, L.R. McDougald, Y.M. Saif (eds). Iowa State University Press, Ames, IA, pp. 541-569.
de Leeuw O, Peeters Β (1999) Complete nucleotide sequence of Newcastle disease virus: evidence for the existence of a new genus within the subfamily Paramyxovirinae. J. Gen. Virol., 80:131-136.
Eash S W, Querbes W, Atwood W J (2004) Infection of Vero cells by BK virus is dependent on caveolae. J. Virol., 78:11583-11590.
Gadaleta P, Vacotto M, Coulombie F (2002) Vesicular stomatitis virus induces apoptosis at early stages in the viral cycle and does not depend on the virus replication. Virus. Res., 86: 87-92.
Georgieva M, Jordanova Ρ (1999) Adaptation of avian reovirus-strain R-85 to a heterologous cell system. Exp. Pathol. Parasitol., 3:45-48.
Govorkova E A, Murti G, Meignier B, De Taisne C, Webster R G (1996) African green monkey kidney (Vero cells) provide an alternative host cell system for influenza A and Β viruses. J. Virol., 70:5519-5524.
Karber G (1931) 50 % end point calculation. Arch. Exp. Pathol. Pharmakol., 162: 480-483.
King D J (1993) Newcastle disease virus passage in MDBK cells as an aid in detection of a virulent subpopulation. Avian Dis., 37: 961-969.
Mohan C M, Dey S, Kumanan Κ, Mahohar Β M, Nainar A M (2007) Adaptation of a velogenic disease virus to Vero cells: assessing the molecular changes before and after infection. Vet. Res. Commun., 31: 371-383.
Nagai Y, Klenk H D, Rott R (1976) Proteolytic cleavage of the viral glycoproteins and its significance for the virulence of Newcastle disease virus. Virology., 72: 494-508.
Nagaleekar V K, Tiwari A K, Kataria R S, Bais M V, Ravindra Ρ V, Kumar S (2007) Bluetongue virus induced apoptosis involves both mitochondrial and death receptor mediated pathways. Arch. Virol., 152:1751-175.
OIE (2004) World organization for Animal Health. Newcastle disease. In: Manual of diagnostic tests and vaccines for terrestrial animals, chapter 2.1.15,5th Ed. OIE, Paris, pp. 270-282.
Peeters Β Ρ, de Leeuw Ο S, Koch G, Gielkens A K (1999) Rescue of Newcastle disease from cloned cDNA: evidence that cleavability of the fusion protein is a major determinant for virulence. J. Virol., 73: 5001-5009.
van Regenmortel, Ranquet C M, Bishop DHL (2000) 7th report of International Committee on Taxonomy of Virus. Academic Press, San Diego, Wien New York. pp. 1024.