傳染性脾臟與腎臟壞死病毒（Infectious Spleen and Kidney Necrosis Virus, ISKNV）為虹彩病毒科（Iridoviridae）巨細胞病毒屬（Megalocytivirus）之雙股DNA病毒，自1990年以來已成為石斑魚、鱸魚等多種養殖魚類的重要病原，常造成大規模死亡，導致嚴重經濟損失。前有研究指出，病毒在感染宿主細胞後，會透過調控宿主之代謝路徑與免疫反應來促進其存活與複製，並改變細胞內代謝物組成，引發氧化壓力、發炎反應與免疫失衡，最終導致細胞死亡。然而，目前對於ISKNV感染魚體後所引起的系統性代謝變化及相關基因調控機制之研究仍相對不足，顯示在病毒防治策略上仍具發展潛力。因此，本研究從多體學角度出發，結合代謝體學與轉錄體學技術，首先應用超高效能液相層析串聯質譜（UHPLC-MS/MS）分析石斑魚感染前後之代謝物變化，以辨識與病毒感染相關的關鍵代謝物；進一步以轉錄體學解析該代謝物所參與之基因調控路徑。結果顯示，病毒感染後與多元不飽和脂肪酸（PUFA）合成相關之代謝途徑明顯受到影響，提示PUFA調控可能為干擾病毒作用的潛在標靶。後續轉錄體分析亦確認PUFA添加具有抑制病毒複製的效果，其作用機制可能與調控抗氧化與免疫發炎相關基因路徑有關。綜上所述，本研究結果提供了以營養干預為核心的抗病毒策略構想，未來有望應用於石斑魚等養殖魚類病毒性疾病之防治。

Infectious spleen and kidney necrosis virus (ISKNV), a double-stranded DNA virus of the Megalocytivirus genus, is a major pathogen in aquaculture, causing high mortality and economic loss in fish such as grouper. ISKNV infection alters host metabolism and immune responses, leading to oxidative stress and inflammation. However, the systemic metabolic and gene regulatory changes in infected fish remain unclear.
This study aimed to investigate ISKNV-induced metabolic alterations and explore potential antiviral strategies using a multi-omics approach. Metabolomic profiling via UHPLC-MS/MS identified key changes in polyunsaturated fatty acid (PUFA) biosynthesis pathways. Subsequent transcriptomic analysis revealed that PUFA supplementation suppressed viral replication by modulating antioxidant and immune-related gene expression.
These findings suggest that PUFA plays a protective role against ISKNV by restoring redox balance and immune homeostasis, highlighting its potential as a nutritional intervention for viral disease control in aquaculture.

漁業署. （2023）. 中華民國漁業統計年報. 行政院農業委員會漁業署.
Bannenberg, G., and Serhan, C.N. Specialized pro-resolving lipid mediators in the inflammatory response: An update. Biochim Biophys Acta 1801, 1260-1273, 2010.
Bazan, N.G. Cellular and molecular events mediated by docosahexaenoic acid-derived neuroprotectin D1 signaling in photoreceptor cell survival and brain protection. Prostaglandins Leukot Essent Fatty Acids 81, 205-211, 2009.
Calder, P.C. Marine omega-3 fatty acids and inflammatory processes: Effects, mechanisms and clinical relevance. Biochim Biophys Acta 1851, 469-484, 2015.
Chen, P.H., Hsueh, T.C., Wu, J.L., and Hong, J.R. Infectious Spleen and Kidney Necrosis Virus (ISKNV) Triggers Mitochondria-Mediated Dynamic Interaction Signals via an Imbalance of Bax/Bak over Bcl-2/Bcl-xL in Fish Cells. Viruses 14, 2022.
Das, U.N. Arachidonic acid and other unsaturated fatty acids and some of their metabolites function as endogenous antimicrobial molecules: A review. J Adv Res 11, 57-66, 2018.
Dong, H.T., Nguyen, V.V., Phiwsaiya, K., Gangnonngiw, W., Withyachumnarnkul, B., Rodkhum, C., and Senapin, S. Concurrent infections of Flavobacterium columnare and Edwardsiella ictaluri in striped catfish, Pangasianodon hypophthalmus in Thailand. Aquaculture 448, 142-150, 2015.
Fiehn, O. Metabolomics--the link between genotypes and phenotypes. Plant Mol Biol 48, 155-171, 2002.
Fusianto, C., Becker, J., Subramaniam, K., Whittington, R., Koda, S., Waltzek, T., Murwantoko, M., and Hick, P. Genotypic Characterization of Infectious Spleen and Kidney Necrosis Virus (ISKNV) in Southeast Asian Aquaculture. Transboundary and Emerging Diseases 2023, 1-16, 2023.
Girisha, S.K., Kushala, K.B., Nithin, M.S., Puneeth, T.G., Naveen Kumar, B.T., Vinay, T.N., Suresh, T., Ajay, S.K., Venugopal, M.N., and Ramesh, K.S. First report of the infectious spleen and kidney necrosis virus (ISKNV) infection in ornamental fishes in India. Transbound Emerg Dis 68, 964-972, 2021.
Golpour, P., Nourbakhsh, M., Mazaherioun, M., Janani, L., Nourbakhsh, M., and Yaghmaei, P. Improvement of NRF2 gene expression and antioxidant status in patients with type 2 diabetes mellitus after supplementation with omega-3 polyunsaturated fatty acids: A double-blind randomised placebo-controlled clinical trial. Diabetes Res Clin Pract 162, 108120, 2020.
Guo, X., Wu, S., Li, N., Lin, Q., Liu, L., Liang, H., Niu, Y., Huang, Z., and Fu, X. Accelerated Metabolite Levels of Aerobic Glycolysis and the Pentose Phosphate Pathway Are Required for Efficient Replication of Infectious Spleen and Kidney Necrosis Virus in Chinese Perch Brain Cells. Biomolecules 9, 440, 2019.
Heemstra, P.C.; Randall, J.E. FAO species catalogue. Vol. 16. Groupers of the world (Family Serranidae, Subfamily Epi- Authors nephelinae). An annotated and illustrated catalogue of the grouper, rockcod, hind, coral grouper and lyretail species known to date. FAO Fisheries Synopsis. No. 125, Vol. 16. Rome, FAO. 1993.382 p., 522 figs, 31 colour plates.
He, J.G., Lü, L., Deng, M., He, H.H., Weng, S.P., Wang, X.H., Zhou, S.Y., Long, Q.X., Wang, X.Z., and Chan, S.M. Sequence analysis of the complete genome of an iridovirus isolated from the tiger frog. Virology 292, 185-197, 2002.
He, J.H., Huang, L., Guo, Z., Weng, S., He, J., and Xu, X. Transcriptional programs of infectious spleen and kidney necrosis virus (ISKNV) in vitro and in vivo. Virus Genes 56, 749-755, 2020.
Hsueh, T.C., Chen, P.H., and Hong, J.R. ISKNV Triggers AMPK/mTOR-Mediated Autophagy Signaling through Oxidative Stress, Inducing Antioxidant Enzyme Expression and Enhancing Viral Replication in GF-1 Cells. Viruses 16, 2024.
Huang, P.-L., Afero, F., Chang, Y., Chen, B.-Y., Lan, H.-Y., Hou, Y.-L., and Huang, C.-T. The Bioeconomic Analysis of Hybrid Giant Grouper (Epinephelus fuscoguttatus × Epinephelus lanceolatus) and Green Grouper (Epinephelus malabaricus): A Case Study in Taiwan. Fishes 8, 610, 2023.
Huang, X., Wei, J., Zheng, Q., Zhang, Y., Zhu, W., Liu, J., Hou, Y., Qin, Q., and Huang, Y. Isolation, identification and genomic analysis of an ISKNV-type megalocytivirus from spotted knifejaw (Oplegnathus punctatus). Aquaculture 532, 736032, 2021.
Inouye, K., Yamano, K., Maeno, Y., Nakajima, K., Matsuoka, M., Wada, Y., and Sorimachi, M. Iridovirus Infection of Cultured Red Sea Bream, Pagrus major. Fish Pathology 27, 19-27, 1992.
Kawsar, M.A., Adikari, D., and Zhang, Y. Autophagy in aquatic animals: mechanisms, implications, and future directions. Front Immunol 16, 1612178, 2025.
Kullgren, A., Jutfelt, F., Fontanillas, R., Sundell, K., Samuelsson, L., Wiklander, K., Kling, P., Koppe, W., Larsson, D.G., Björnsson, B.T., and Jönsson, E. The impact of temperature on the metabolome and endocrine metabolic signals in Atlantic salmon (Salmo salar). Comp Biochem Physiol A Mol Integr Physiol 164, 44-53, 2013.
Kurita, J., Nakajima, K., Hirono, I., and Aoki, T. Polymerase Chain Reaction (PCR) Amplification of DNA of Red Sea Bream Iridovirus (RSIV). Fish Pathology 33, 17-23, 1998.
Leu, J.H., Yang, F., Zhang, X., Xu, X., Kou, G.H., and Lo, C.F. Whispovirus. Curr Top Microbiol Immunol 328, 197-227, 2009.
Li, P.H., Cai, Y.J., Zhu, X.L., Yang, J.D., Yang, S.Q., Huang, W., Wei, S.N., Zhou, S., Wei, J.G., Qin, Q.W., and Sun, H.Y. Epinephelus coioides Hsp27 negatively regulates innate immune response and apoptosis induced by Singapore grouper iridovirus (SGIV) infection. Fish Shellfish Immunol 120, 470-480, 2022.
Liu, H.I., Chiou, P.P., Gong, H.Y., and Chou, H.Y. Cloning of the Major Capsid Protein (MCP) of Grouper Iridovirus of Taiwan (TGIV) and Preliminary Evaluation of a Recombinant MCP Vaccine against TGIV. Int J Mol Sci 16, 28647-28656, 2015.
Lü, L., Zhou, S.Y., Chen, C., Weng, S.P., Chan, S.M., and He, J.G. Complete genome sequence analysis of an iridovirus isolated from the orange-spotted grouper, Epinephelus coioides. Virology 339, 81-100, 2005.
Magnadóttir, B. Innate immunity of fish (overview). Fish Shellfish Immunol 20, 137-151, 2006.
McNamara, R.K. Role of Omega-3 fatty acids in the etiology, treatment, and prevention of depression: Current status and future directions. Journal of Nutrition & Intermediary Metabolism 5, 96-106, 2016.
Nicholson, J.K., Lindon, J.C., and Holmes, E. 'Metabonomics': understanding the metabolic responses of living systems to pathophysiological stimuli via multivariate statistical analysis of biological NMR spectroscopic data. Xenobiotica 29, 1181-1189, 1999.
Pattanayak, S., Paul, A., and Sahoo, P.K. Detection and genetic analysis of infectious spleen and kidney necrosis virus (ISKNV) in ornamental fish from non-clinical cases: First report from India. bioRxiv, 2020.2008.2012.247650, 2020.
Patti, G.J., Yanes, O., and Siuzdak, G. Innovation: Metabolomics: the apogee of the omics trilogy. Nat Rev Mol Cell Biol 13, 263-269, 2012.
Russell, R.C., Tian, Y., Yuan, H., Park, H.W., Chang, Y.Y., Kim, J., Kim, H., Neufeld, T.P., Dillin, A., and Guan, K.L. ULK1 induces autophagy by phosphorylating Beclin-1 and activating VPS34 lipid kinase. Nat Cell Biol 15, 741-750, 2013.
Sargent, J.R., Tocher, D.R., and Bell, J.G. 4 - The Lipids. In Fish Nutrition (Third Edition), J.E. Halver, and R.W. Hardy, eds. (San Diego: Academic Press), pp. 181-257. 2003.
Simon, C., and Daniel, R. Metagenomic analyses: past and future trends. Appl Environ Microbiol 77, 1153-1161, 2011.
Tocher, D. Fatty acid requirements in ontogeny of marine and freshwater fish. Aquaculture Research 41, 2010.
Tocher, D.R. Metabolism and Functions of Lipids and Fatty Acids in Teleost Fish. Reviews in Fisheries Science 11, 107-184, 2003.
Torstensen, B.E., Bell, J.G., Rosenlund, G., Henderson, R.J., Graff, I.E., Tocher, D.R., Lie, Ø., and Sargent, J.R. Tailoring of Atlantic salmon (Salmo salar L.) flesh lipid composition and sensory quality by replacing fish oil with a vegetable oil blend. J Agric Food Chem 53, 10166-10178, 2005.
Wang, S., Huang, X., Huang, Y., Hao, X., Xu, H., Cai, M., Wang, H., and Qin, Q. Entry of a novel marine DNA virus, Singapore grouper iridovirus, into host cells occurs via clathrin-mediated endocytosis and macropinocytosis in a pH-dependent manner. J Virol 88, 13047-13063, 2014.
Williams, T., Barbosa Solomieu, V., and Chinchar, V. A Decade of Advances in Iridovirus Research. Advances in virus research 65, 173-248, 2005.
Wu, J.L., Nishioka, T., Mori, K., Nishizawa, T., and Muroga, K. A time-course study on the resistance of Penaeus japonicus induced by artificial infection with white spot syndrome virus. Fish Shellfish Immunol 13, 391-403, 2002.
Xia, J., Sinelnikov, I.V., Han, B., and Wishart, D.S. MetaboAnalyst 3.0--making metabolomics more meaningful. Nucleic Acids Res 43, W251-257, 2015.
Yan, Y., Cui, H., Jiang, S., Huang, Y., Huang, X., Wei, S., Xu, W., and Qin, Q. Identification of a novel marine fish virus, Singapore grouper iridovirus-encoded microRNAs expressed in grouper cells by Solexa sequencing. PLoS One 6, e19148, 2011.
Zhou, Y., Xu, C., Gu, S., Xiao, Y., Wu, S., Wang, H., and Bao, W. Integrated Metabolomic and transcriptomic analyses reveal deoxycholic acid promotes transmissible gastroenteritis virus infection by inhibiting phosphorylation of NF-κB and STAT3. BMC Genomics 25, 239, 2024.