Indian Journal of Animal Research 930 RESEARCH ARTICLE Indian Journal of Animal Research, Volume 55 Issue 8: 930-935 (August 2021) Pathological and Molecular Characterization of Newcastle Disease Virus Isolated from Gallus gallus in Java, Indonesia A.N.M. Ansori 1,2 , M.K.J. Kusala 1,2 , R.V. Nidom 2,3 , S. Indrasari 2,3 , K. Zarkasie 4 , K.P. Santoso 1,2 , C.A. Nidom 1,2,3 10.18805/IJAR.B-1301 ABSTRACT Background: Known as the vital avian disease in Indonesia, Newcastle disease (ND) has surged all over Indonesia which cause an immense drop in poultry industry. This investigation intended to examine the pathological and molecular characteristics of the latest Newcastle disease virus (NDV) derived from Gallus gallus in Java, Indonesia. Methods: The samples were inoculated on the embryonic chicken eggs (ECEs), then identified using the hemagglutination test (HA test), insulated isothermal PCR (iiPCR) and hemagglutination inhibition test (HI test). It conducted the reverse transcription-polymerase chain reaction (RT-PCR) using MSF1 and MSF2 primers and followed by nucleotide sequencing for the fusion protein gene. B-cells epitope prediction analysis was applied using the Immune Epitope Database and Analysis Resource (IEDB) and VaxiJen v2.0 to predict its immunogenicity. In addition, it also performed the histopathological examination by applying hematoxylin and eosin staining. Result: The results identified two viruses, NDV-1 and NDV-2, based on the fusion protein gene. Significantly, the cleavage site motif of NDV1 is 112 GRQGRL 117 (avirulent) and whereas for NDV2 is 112 RRRKRF 117 (virulent). Furthermore, it also predicted the “CKMGSRPSTKNPAP” peptide from NDV1 could be an immunogenic epitope candidate with a BepiPred linear epitope prediction score of 17.08. In summary, it suggests that NDV1 isolate can be used as a reference for vaccine design in Indonesia. Key words: Avian paramyxovirus type-1, Fusion protein gene, Molecular characterization, Newcastle disease, Pathological. INTRODUCTION To meet the required amount and quality of animal protein, eggs and chicken can be the primary source for this need. Consequently, people would like to fulfill their protein needs with those sources which brings a large impact on poultry farming like in Indonesia. As its meat and eggs are inexpensive supply of animal protein widely available globally, thus, the poultry industry is a fundamental provider to global food supply. In 2013, the United States was ranked first as a producer of poultry meat, whereas in Asia, Indonesia was in the fourth position after China, India and Iran (Wahyono and Utami, 2018). Avian paramyxovirus serotype-1 (APMV-1), also commonly known as NDV caused ND. ND has the possibility to cause catastrophic losses, due to the fact that it is an extremely contagious and deadly disease in poultry industry and is an A listed disease categorized by the Office International des Epizooties (OIE) (Ganar et al., 2014; Chowdhary et al., 2020). Affliction by a virulent NDV is associated with paralysis, neurological, respiratory and high mortality in poultry (Mohanambal et al., 2018). Furthermore, ND also can determine any economic losses due to the death, growth retardation and decreased egg production, as well as other losses connected with post-vaccination responses (Cornax et al., 2012). One of the attempts to fight against NDV is through a vaccination program (Ganar et al., 2014). Moreover, NDV has a single serotype (APMV-1) and a genome size of about 15.2 kb which is single stranded, not segmented, negative-sense RNA genomes that encode nucleocapsid protein (NP), fusion protein (F), phosphoproteins (P), RNA-dependent RNA polymerases or RdRps (L), matrix protein (M), hemagglutinin-neuraminidase protein (HN) and two collateral nonstructural proteins (V and W) (Ganar et al., 2014). Among the aforementioned proteins, F protein is the main target of the immune response and has the high immunogenic properties (Putri et al., 2021). NDV can infect almost all species in poultry industry, both wild and domesticated species. In addition, ND outbreaks have occurred in almost all countries in the world (Ewies et al., 2017; Nath and Kumar, 2017), including Indonesia (Putri et al., 2021). Vaccinology Laboratory, Professor Nidom Foundation, Surabaya, Indonesia. 1 Faculty of Veterinary Medicine, Universitas Airlangga, Surabaya, Indonesia. 2 Professor Nidom Foundation, Surabaya, Indonesia. 3 PT. Riset AIRC Indonesia, Surabaya, Indonesia. 4 PT. Global Biosains Teknologi, Bogor, Indonesia. Corresponding Author: C.A. Nidom, Faculty of Veterinary Medicine, Universitas Airlangga, Surabaya, Indonesia. Email: nidomca@fkh.unair.ac.id; nidomca@pnfinstitute.org How to cite this article: Ansori, A.N.M., Kusala, M.K.J., Nidom, R.V., Indrasari, S., Zarkasie, K., Santoso, K.P. and Nidom, C.A. (2021). Pathological and Molecular Characterization of Newcastle Disease Virus Isolated from Gallus gallus in Java, Indonesia. Indian Journal of Animal Research. 55(8): 930-935. DOI: 10.18805/ IJAR.B-1301. Submitted: 16-06-2020 Accepted: 15-04-2021 Online: 30-04-2021