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Received: Revised: Accepted: Published online: October 10, 2018 February 13, 2019 February 28, 2019 March 07, 2019 In Pakistan, growth of poultry industry is confronted by many viral diseases and Newcastle Disease (ND) is on top. The present study was conducted for pathotyping and genetic characterization of NDV isolated from an outbreak in District Okara region of Punjab, Pakistan. Post mortem examination of fresh carcasses (n=10) was performed and various organs like trachea, spleen, lung tissues, and cloacal contents were collected from birds showing typical lesions of the disease aseptically. Tissue homogenate was inoculated in 9 days old embryonated chicken eggs and allantoic fluid was harvested. The presence of virulent virus was checked by hemagglutination, hemagglutination inhibition using hyperimmune serum and ICPI assays. NDV was purified using sucrose gradient ultracentrifugation and confirmation was done by F-gene amplification, SDS-PAGE and mass spectrometry of fusion protein. The HA titer was 1:256, and hemagglutination inhibition was achieved using Anti NDV serum. A band of ≈ 800 bp of F gene (Acc. No. Okara/Pakistan/MH607122) was amplified which revealed 99% similarity with UVAS/Pak/2015/MF437287, UVAS/Pak/2016/KX791187 and Tehran/Iran/MG871466 strains. SDS-PAGE yielded five bands including Fusion protein with the estimated mass of 58896.0 Da. The presence of fusion protein and ICPI value was 2 which confirmed virulence of isolated strain resulting in high mortality. Thus, it is the need of the hour to characterize the prevalent indigenous strains of NDV to develop some novel vaccine strategies for effective control. ©2019 PVJ. All rights reserved

Pathotyping and Genetic Characterization of a Highly Virulent Newcastle Disease Virus Strain Isolated from Recent Outbreak in District Okara, Pakistan