ABSTRACT. To provide information on the epidemiology of Newcastle disease (ND) of poultry in Taiwan, ND virus isolates from chickens and an owl were investigated by restriction site analysis and sequencing of their gene. A 1,349 base fragment of the F (fusion protein) gene was amplified by reverse transcription-polymerase chain reaction (RT-PCR). The PCR products were analyzed using restriction endonucleases, HinfI, BstOI, and RsaI. Three strains isolated from chickens during the 1995 epidemic outbreak had the same restriction sites as that of a 1994 isolate; the number of the restriction sites of HinfI, BstOI, and RsaI were 4, 2, and 4, respectively. In the F gene of the strain isolated from an owl during the same outbreak an additional restriction site of HinfI was found. The 1991 isolate had only 3 restriction sites. The F gene of the owl isolate was amplified by RT-PCR and followed by direct sequencing. The deduced amino acid sequence at the cleavage site of the F protein was of virulent strains, 112 R-R-Q-K-R-F 117 . The F gene of Ow/Tw/2209/95 was phylogenetically most closely related to that of Ck/Tw/2137/95 isolated from the same outbreak. The present results indicate that the causative virus of the 1995 ND outbreak had already been present in Taiwan.-KEY WORDS: chicken, F gene, Newcastle disease, owl, RFLP.J. Vet. Med. Sci. 61(11): 1191-1195, 1999 possibility, restriction site analysis for the F gene was applied to the NDV strains isolated in Taiwan before and during the 1995 outbreak. The F gene of the NDV isolate (Ow/Tw/2209/95) from a barrel-toe scop owl (Otus bakkamoena glabripes) was sequenced. The nucleotide sequence was compared with those of several foreign NDV strains to investigate the evolution of Taiwan isolates. The results indicate the causative virus originated from one that had been present in Taiwan before the 1995 outbreak.
MATERIALS AND METHODS
Viruses and pathogenicity tests:The six NDV strains used in this study were isolated from sporadic cases before the 1995 and during the 1995 outbreak in Taiwan. Chicken/ Taiwan/970/1991 (Ck/Tw/970/91) and Ck/Tw/2066/94 were isolated before the 1995 outbreak, and Ck/Tw/2137/95 (Taiwan95, Accession number: U62620, GenBank), Ck/Tw/ 2164/95, and Ck/Tw/2174/95 were isolated from chickens during the 1995 outbreak. Owl (Ow)/Tw/2209/95 was isolated from a dead owl in Taipei City Zoo in April 1995 [13]. A virulent strain (Sato) used at the Taiwan Animal Health Research Institute (TAHRI, Tansui) for ND vaccine potency test as the challenge strain was also used.After inoculation of the sample filtrates to SPF chicken embryos (TAHRI), viruses were recovered from the allantoic cavity and confirmed by electron microscopy and the hemagglutination-inhibition test with specific anti-NDV antiserum (produced in SPF chickens, TAHRI). The pathogenicity of the isolates was determined by the intracerebral pathogenicity index (ICPI) and intravenous pathogenicity index (IVPI) [1].RNA extraction: All glassware and double distilled water Newcastle disease (ND) is an econ...