PERTANIKA JOURNAL OF SCIENCE AND TECHNOLOGY

Inclusion body hepatitis (IBH) is considered one of the re-emerging diseases of avian virus that causes economic damage worldwide. IBH is caused by different serotypes of fowl adenovirus (FAdV), and most of the FAdV cases in Malaysia are related to the serotype 8b. The objective of this study was to determine the molecular markers of UPMT27 Malaysian FAdV isolate and to identify the evolutionary relationship through the phylogenetic approach. Propagation of the isolate was made in embryonated chicken eggs and chicken embryo liver (CEL cells) before it was subjected to viral DNA extraction. Both the fiber and hexon genes of the isolate were amplified and sequenced. The sequences were aligned with the published FAdV sequences. The results showed 100% identity between UPMT27 and the previous Malaysian isolates. A phylogenetic study showed that UPMT27 was closely related to the previous Malaysian isolates. Interestingly, the substitution of the amino acids was consistent between the Malaysia isolates of both fiber protein at positions 72 (Serine ─serine), 101 (Alanine -alanine), 125 (Glycine-glycine), and hexon protein 85 (Serine-serine) 160 (Glutamate- glutamate) and 205 (Alanine-alanine) respectively. It appeared that the amino acid variations were the indicators for genetic diversity. Thus, these findings provide information on the evolutionary relationship between FAdV subtypes for IBH prevention.

• Chiocca, S., Kurzbauer, R., Schaffner, G., Baker, A., Mautner, V., & Cotten, M. (1996). The complete DNA sequence and genomic organization of the avian adenovirus CELO. Journal of Virology, 70(5), 2939-2949.

• Grgić, H., Yang, D. H., & Nagy, É. (2011). Pathogenicity and complete genome sequence of a fowl adenovirus serotype 8 isolate. Virus Research, 156(1-2), 91-97. doi: https://doi.org/10.1016/j.virusres.2011.01.002

• Grgić, H., Poljak, Z., Sharif, S., & Nagy, É. (2013). Pathogenicity and cytokine gene expression pattern of a serotype 4 fowl adenovirus isolate. PloS One, 8(10), 1-10. doi: https://doi.org/10.1371/journal.pone.0077601

• Grgić, H., Krell, P. J., & Nagy, É. (2014). Comparison of fiber gene sequences of inclusion body hepatitis (IBH) and non-IBH strains of serotype 8 and 11 fowl adenoviruses. Virus Genes, 48(1), 74-80.

• Gupta, A., Popowich, S., Ojkic, D., Kurukulasuriya, S., Chow-Lockerbie, B., Gunawardana, T., ... & Tikoo, S. K. (2018). Inactivated and live bivalent fowl adenovirus (FAdV8b+ FAdV11) breeder vaccines provide broad-spectrum protection in chicks against inclusion body hepatitis (IBH). Vaccine, 36(5), 744-750. doi: https://doi.org/10.1016/j.vaccine.2017.12.047

• Hair-Bejo, M. (2005). Inclusion body hepatitis in a flock of commercial broiler chickens. Journal of Veterinary Malaysia, 17(1), 23–26.

• Huang, Q., Ma, X., Huang, X., Huang, Y., Yang, S., Zhang, L., ... & Xu, C. (2019). Pathogenicity and complete genome sequence of a fowl adenovirus serotype 8b isolate from China. Poultry Science, 98(2), 573-580. doi: https://doi.org/10.3382/ps/pey425

• James, N. M., & Edward, J. D. (2011). Fenner’s veterinary viriology (4th Edition). San Diego, USA: Academic Press.

• Juliana, M. A., Nurulfiza, M. I., Hair-Bejo, M., Omar, A. R., & Aini, I. (2014). Molecular characterization of fowl adenoviruses isolated from inclusion body hepatitis outbreaks in commercial broiler chickens in Malaysia. Pertanika Journal of Tropical Agriculture Science, 37(4), 483-497.

• Li, P. H., Zheng, P. P., Zhang, T. F., Wen, G. Y., Shao, H. B., & Luo, Q. P. (2017). Fowl adenovirus serotype 4: Epidemiology, pathogenesis, diagnostic detection, and vaccine strategies. Poultry Science, 96(8), 2630-2640. doi: https://doi.org/10.3382/ps/pex087

• Mansoor, M. K., Hussain, I., Arshad, M., & Muhammad, G. (2011). Preparation and evaluation of chicken embryo-adapted fowl adenovirus serotype 4 vaccine in broiler chickens. Tropical Animal Health and Production, 43(2), 331-338. doi: https://doi.org/10.1007/s11250-010-9694-z

• Majdi, A., & Hair-Bejo, M. (2015, February 23-27). Pathogenicity of Malaysian fowl adenovirus isolates in specific pathogen free chickens. In 10th Proceedings of the Seminar of Veterinary Sciences (pp. 15-20). Faculty of Veterinary Medicine, Universiti Putra Malaysia, Malaysia.

• Marek, A., Kaján, G. L., Kosiol, C., Benkő, M., Schachner, A., & Hess, M. (2016). Genetic diversity of species Fowl aviadenovirus D and Fowl aviadenovirus E. Journal of General Virology, 97(9), 2323-2332. doi: https://doi.org/10.1099/jgv.0.000519

• Marek, A., Kosiol, C., Harrach, B., Kajan, G. L., Schlotterer, C., & Hess, M. (2013). The first whole genome sequence of a fowl adenovirus B strain enables interspecies comparisons within the genus aviadenovirus. Journal Veterinary Microbiology, 166(1-2), 250-256. doi: https://doi.org/10.1016/j.vetmic.2013.05.017

• Marek, A., Schulz, E., Hess, C., & Hess, M. (2010). Comparison of the fibers of fowl adenovirus A serotype 1 isolates from chickens with gizzard erosions in Europe and a pathogenic reference strains. Journal of Veterinary Diagnostics, 22(6), 937-941. doi: https://doi.org/10.1177/104063871002200613

• Marek, A., Nolte, V., Schachner, A., Berger, E., Schlötterer, C., & Hess, M. (2012). Two fiber genes of nearly equal lengths are a common and distinctive feature of Fowl adenovirus C members. Veterinary Microbiology, 156(3-4), 411-417. doi: https://doi.org/10.1016/j.vetmic.2011.11.003

• McFerran, J. B., & Smyth, J. A. (2000). Avian adenoviruses. Revue Scientifique Et Technique (International Office of Epizootics), 19(2), 589-601.

• Meulemans, G., Couvreur, B., Decaesstecker, M., Boschmans, M., & van den Berg, T. P. (2004). Phylogenetic analysis of fowl adenoviruses. Journal of Avian Pathology, 33(2), 164-170. doi: https://doi.org/10.1080/03079450310001652086

• Morshed, R., Hosseini, H., Langeroude, A. G., Fard, M. H. B., & Charkhkar, S. (2017). Fowl adenoviruses D and E cause inclusion body hepatitis outbreaks in broilers and broilers breeder pullet flocks. Avian Disease, 61(2), 205-210. doi: https://doi.org/10.1637/11551-120516-Reg.1

• Niu, Y. J., Sun, W., Zhang, G. H., Qu, Y. J., Wang, P. F., Sun, H. L., & Liu, S. D. (2016). Hydropericardium syndrome outbreak caused by fowl adenovirus serotype 4 in China. Journal of General Virology, 97, 2684-2690. doi: https://doi.org/10.1099/jgv.0.000567

• Norfitriah, M. S. (2018). Development of live attenuated Fowl adenovirus isolate of Malaysia for future production of vaccine (PhD Thesis). Universiti Putra Malaysia, Malaysia.

• Norfitriah, M. S., Hair-Bejo. M., Omar, A. R., Aini, I., & Nurulfiza, M. I. (2018). Molecular detection and pathogenicity of fowl adenovirus isolated from disease outbreak in commercial layer farm. International Journal of Agriculture Sciences and Veterinary Medicine, 6(1), 73-84.

• Norina, L., Norsharina, A., Nurnadiah, A., Redzuan, I., Ardy, A., & Nor-Ismaliza, I. (2016). Avian adenovirus isolated from broiler affected with Inclusion body hepatitis. Malaysian Journal of Veterinary Research, 7(2), 121-126.

• Ojkic, D., & Nagy, É. (2000). The complete nucleotide sequence of fowl adenovirus type 8. Journal of General Virology, 81(7), 1833-1837.doi: https://doi.org/10.1099/0022-1317-81-7-1833

• Pallister, J., Wright, P. J., & Sheppard, M. (1996). A single gene encoding the fiber is responsible for variations in virulence in the fowl adenoviruses. Journal of Virology, 70(8), 5115-5122.

• Popowich, S., Gupta, A., Chow-lockerbie, B., Ayalew, L., Ambrose, N., Ojkic, D., … & Suresh, K. (2018). Broad spectrum protection of broiler chickens against inclusion body hepatitis by immunizing their broiler breeder parents with a bivalent live fowl adenovirus vaccine. Research in Veterinary Science, 118, 262-269. doi: https://doi.org/10.1016/j.rvsc.2018.03.003

• Rux, J. J., & Burnett, R. M. (2003). Structural and phylogenetic analysis of adenovirus hexons by use of high-resolution X-ray crystallographic, molecular modeling, and sequence based methods. Journal of Virology, 77, 9553-9566. doi: 10.1128/JVI.77.17.9553-9566.2003

• Schachner, A., Gonzalez, G., Endler, L., Ito, K., & Hess, M. (2019). Fowl Adenovirus (FAdV) Recombination with Intertypic Crossovers in Genomes of FAdV-D and FAdV-E, Displaying Hybrid Serological Phenotypes. Viruses, 11(12), 1-24. doi: https://doi.org/10.3390/v11121094

• Shah, M. S., Ashraf, A., Khan, M. I., Rahman, M., Habib, M., Chughtai, M. I., & Qureshi, J. A. (2017). Fowl adenovirus: history, emergence, biology and development of a vaccine against hydropericardium syndrome. Archives of Virology, 162(7), 1833-1843. doi: https://doi.org/10.1007/s00705-017-3313-5

• Slaine, P. D., Ackford, J. G., Kropinski, A. M., Kozak, R. A., Krell, P. J., & Nagy, E. (2016). Molecular characterization of pathogenic and non-pathogenic fowl aviadenovirus serotype 11 isolates. Journal of Microbiology, 62(12), 993-1002. doi: https://doi.org/10.1139/cjm-2016-0297

• Soumyalekshmi, S., Ajith, M. K., & Chandraprakash, M. (2014). Isolation of fowl adenovirus in chicken embryo liver cell culture and its detection by hexon gene based PCR. Journal of Science Research and Technology, 2(3), 33-36.

• Wang, Z., & Zhao, J. (2019). Pathogenesis of Hypervirulent Fowl Adenovirus Serotype 4: The Contributions of Viral and Host Factors. Viruses, 11(8), 1-10. doi: https://doi.org/10.3390/v11080741

• Zhang, Y., Liu, R., Tian, K., Wang, Z., Yang, X., Gao, D., ... & Zhao, J. (2018). Fiber2 and hexon genes are closely associated with the virulence of the emerging and highly pathogenic fowl adenovirus 4. Emerging Microbes and Infections, 7(1), 1-10. doi: https://doi.org/10.1038/s41426-018-0203-1

• Zhao, J., Zhong, Q. I., Zhao, Y., Hu, Y. X., & Zhang, G. Z. (2015). Pathogenicity and complete genome characterization of fowl adenoviruses isolated from chickens associated with inclusion body hepatitis and hydropericardium syndrome in China. PloS One, 10(7), 1-14. doi: https://doi.org/10.1371/journal.pone.0133073