caa a22 4500 463237366 CHVBK 20180405153302.0 cr unu---uuuuu 170326e20070401xx s 000 0 eng 10.1007/s11259-006-3502-2 doi (NATIONALLICENCE)springer-10.1007/s11259-006-3502-2 Adaptation of a Velogenic Newcastle Disease Virus to Vero Cells: Assessing the Molecular Changes Before and After Adaptation [Elektronische Daten] [C. Mohan, Sohini Dey, K. Kumanan, B. Manohar, A. Nainar] A velogenic Newcastle disease virus isolate was passaged 50 times in Vero cell culture and the virus was assessed for the molecular changes associated with the passaging. At every 10th passage, the virus was characterized conventionally by mean death time (MDT) analysis, intracerebral pathogenicity index (ICPI) and virus titration. At increasing passage levels, a gradual reduction in the virulence of the virus was observed. Molecular characterization of the virus included cloning and sequencing of a portion of the fusion gene (1349 bp) encompassing the fusion protein cleavage site (FPCS), which was previously amplified by reverse transcription-polymerase chain reaction. Sequence analysis revealed a total of 135 nucleotide substitutions which resulted in the change of 42 amino acids between the velogenic virus and the 50th passage virus. The predicted amino acid motif present at the cleavage site of the virulent virus was 109SRRRRQRRFVG119 and the corresponding region of the adapted adapted virus was 109SGGRRQKRFIG119. Pathogenicity studies conducted in 20-week-old seronegative birds revealed gross lesions such as petechial haemorrhages in the trachea, proventricular junction and intestines, and histopathological changes such as depletion and necrosis of the lymphocytes in thymus, spleen, bursa and caecal tonsils in the birds injected with the velogenic virus and absence of the lesions in birds injected with the adapted virus.The 50th-passage cell culture virus was back-passaged five times in susceptible chickens and subjected to virulence attribute analysis and sequence analysis of the FPCS region, with minor difference found between them. Springer Science + Business Media, Inc., 2007 Newcastle disease virus nationallicence Vero cells nationallicence fusion gene nationallicence sequencing studies nationallicence pathogenicity testing nationallicence AAF : amnioallantoic fluid nationallicence CPE(s) : cytopathic effect(s) nationallicence dNTPs : deoxynucleotide triphosphate nationallicence ECE : embryonated chicken eggs nationallicence FPCS : fusion protein cleavage site nationallicence HA : haemagglutination nationallicence HI : haemagglutination inhibition nationallicence ICPI : intracerebral pathogenicity index nationallicence MDT : mean death time nationallicence MuMLV : Moloney murine leukemia virus nationallicence ND : Newcastle disease nationallicence NDV : Newcastle disease virus nationallicence p.i. : post inoculation nationallicence RT-PCR : reverse transcription-polymerase chain reaction nationallicence TCID50 : tissue culture infective dose 50% nationallicence Mohan C. Department of Animal Biotechnology, Madras Veterinary College, Chennai aut Dey Sohini Department of Animal Biotechnology, Madras Veterinary College, Chennai aut Kumanan K. Department of Animal Biotechnology, Madras Veterinary College, Chennai aut Manohar B. Department of Animal Biotechnology, Madras Veterinary College, Chennai aut Nainar A. Department of Animal Biotechnology, Madras Veterinary College, Chennai aut Veterinary Research Communications Kluwer Academic Publishers 31/3(2007-04-01), 371-383 0165-7380 31:3<371 2007 31 11259 https://doi.org/10.1007/s11259-006-3502-2 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s11259-006-3502-2 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Mohan C. Department of Animal Biotechnology, Madras Veterinary College, Chennai aut NATIONALLICENCE 700 1- Dey Sohini Department of Animal Biotechnology, Madras Veterinary College, Chennai aut NATIONALLICENCE 700 1- Kumanan K. Department of Animal Biotechnology, Madras Veterinary College, Chennai aut NATIONALLICENCE 700 1- Manohar B. Department of Animal Biotechnology, Madras Veterinary College, Chennai aut NATIONALLICENCE 700 1- Nainar A. Department of Animal Biotechnology, Madras Veterinary College, Chennai aut NATIONALLICENCE 773 0- Veterinary Research Communications Kluwer Academic Publishers 31/3(2007-04-01), 371-383 0165-7380 31:3<371 2007 31 11259 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer