EFFECTS OF FETAL BOVINE SERUM CONCENTRATION AND VENTED FLASK CONDITIONS ON E-11 CELL GROWTH AND TILAPIA LAKE VIRUS PROPAGATION
DOI:
https://doi.org/10.24191/joa.v14i1.9648Keywords:
Aquaculture, Tilapia Lake Virus (TiLV), E-11 cells, Cytopathic effectAbstract
Tilapia Lake Virus (TiLV) is an emerging pathogen threatening global tilapia aquaculture, causing high mortality and significant economic losses. The present study aimed to optimize E-11 cell culture conditions for improved growth and TiLV propagation under different serum and flask ventilation conditions. E-11 cells were cultured in Leibovitz’s L-15 medium supplemented with 10%, 15%, and 20% fetal bovine serum (FBS) and maintained at 25°C in a CO₂-free incubator. Cell viability was assessed over four days using the Trypan Blue exclusion assay, while TiLV propagation was evaluated in both seal-cap and vent-cap flasks through microscopic observation of cytopathic effects (CPE). Results showed that E-11 cells supplemented with 15% FBS exhibited the highest viability (85–90%) and optimal proliferation compared to cells grown in 10% or 20% FBS. Morphologically, all cultures displayed similar epithelial-like characteristics with polygonal cell shapes and intact monolayers under the microscope. No significant differences in viability were observed between open- and closed-vented flasks; however, CPE were detected only in infected E-11 cells maintained in vent-cap flasks by 10 days post-infection (dpi), while cells in seal-cap flasks remained morphologically intact. These findings suggest that a 15% FBS concentration and a vented culture environment support both healthy E-11 cell growth and effective TiLV propagation. This optimized culture system enhances the reliability of in vitro TiLV studies and supports efforts toward sustainable aquaculture and food security.
References
Aarattuthodi, S., Dharan, V., & Koshy, M. (2022). Fish Cell Cultures - Uses and Prospects. Journal of Aquaculture Research and Development, 13(1), 667.
Aich, N., Paul, A., Choudhury, T. G., & Saha, H. (2022). Tilapia Lake Virus (TiLV) disease: Current status of understanding. Aquaculture and Fisheries, 7(1), 7–17. https://doi.org/10.1016/j.aaf.2021.04.007
Goswami, M., Yashwanth, B. S., Trudeau, V., & Lakra, W. S. (2022). Role and relevance of fish cell lines in advanced in vitro research. Molecular Biology Reports, 49(3), 2393–2411. https://doi.org/10.1007/s11033-021-06997-4
Lazaro, T. M., de Alcântara Rocha, N. R., Levy-Pereira, N., & de Sousa, R. L. M. (2024). Essential features of in vitro fish cell culture: an overview. Latin American Journal of Aquatic Research , 52(2), 196–208. https://doi.org/10.3856/vol52-issue2-fulltext-3131
Leland, D. S., & Ginocchio, C. C. (2007). Role of cell culture for virus detection in the age of technology. Clinical Microbiology Reviews, 20(1), 49–78. https://doi.org/10.1128/CMR.00002-06
Mugimba, K. K., Tal, S., Dubey, S., Mutoloki, S., Dishon, A., Evensen, A., & Munang’andu, H. M. (2019). Gray (Oreochromis niloticus X O. aureus) and red (Oreochromis spp.) tilapia show equal susceptibility and proinflammatory cytokine responses to experimental tilapia lake virus infection. Viruses, 11(10). https://doi.org/10.3390/v11100893
Ozawa, Y., Hashizume, T., & Ying, B. W. (2024). A data-driven approach for cell culture medium optimization. Biochemical Engineering Journal, 214, 109591. https://doi.org/10.1016/j.bej.2024.109591
Piamsomboon, P., & Wongtavatchai, J. (2021). Detection of tilapia lake virus (TiLV) in healthy fish from the pre-existing disease environment using different RT-PCR methods. Turkish Journal of Fisheries and Aquatic Sciences, 21(4), 205–209. https://doi.org/10.4194/1303-2712-v21_4_05
Pierezan, F., Yun, S., Piewbang, C., Surachetpong, W., & Soto, E. (2020). Pathogenesis and immune response of Nile tilapia (Oreochromis niloticus) exposed to Tilapia lake virus by intragastric route. Fish and Shellfish Immunology, 107(Pt A), 289–300. https://doi.org/10.1016/j.fsi.2020.10.019
Rogers, Z. J., Flood, D., & Taylor, C. T. (2024). Oxygen control in cell culture – Your cells may not be experiencing what you think!. Free Radical Biology and Medicine, 226, 279–287. https://doi.org/10.1016/j.freeradbiomed.2024.11.036
Segeritz, C., & Vallier, L. (2017). Cell culture : Growing cells as model systems in vitro. Basic Science Methods for Clinical Researchers, 151-172. Academic Press. https://doi.org/10.1016/B978-0-12-803077-6.00009-6
Surachetpong, W., Roy, S. R. K., & Nicholson, P. (2020). Tilapia lake virus: The story so far. Journal of Fish Diseases, 43(10), 1115–1132. https://doi.org/10.1111/jfd.13237
Tattiyapong, P., Dachavichitlead, W., & Surachetpong, W. (2017). Experimental infection of Tilapia Lake Virus (TiLV) in Nile tilapia (Oreochromis niloticus) and red tilapia (Oreochromis spp.). Veterinary Microbiology, 207, 170–177. https://doi.org/10.1016/j.vetmic.2017.06.014
Uysal, O., Sevimli, T., Sevimli, M., Gunes, S., & Sariboyaci, A. E. (2018). Cell and tissue culture: The base of biotechnology. Omics Technologies and Bio-engineering, (391-429). Academic Press. https://doi.org/10.1016/B978-0-12-804659-3.00017-8
Weiskirchen, S., Schröder, S. K., Buhl, E. M., & Weiskirchen, R. (2023). A beginner’s guide to cell culture: Practical advice for preventing needless problems. Cells, 12(5). https://doi.org/10.3390/cells12050682
Widziolek, M., Janik, K., Mojzesz, M., Pooranachandran, N., Adamek, M., Pecio, A., Surachetpong, W., Levraud, J. P., Boudinot, P., Chadzinska, M., & Rakus, K. (2021). Type I interferon-dependent response of zebrafish larvae during tilapia lake virus (TiLV) infection. Developmental and Comparative Immunology, 116, 103936. https://doi.org/10.1016/j.dci.2020.103936
Yamkasem, J., Tattiyapong, P., Kamlangdee, A., & Surachetpong, W. (2019). Evidence of potential vertical transmission of tilapia lake virus. Journal of Fish Diseases, 42(9), 1293–1300. https://doi.org/10.1111/jfd.13050
Yu, N. tong, Zeng, W. wei, Xiong, Z., & Liu, Z. xin. (2022). A high efficacy DNA vaccine against Tilapia lake virus in Nile tilapia (Oreochromis niloticus). Aquaculture Reports, 24, 101166. https://doi.org/10.1016/j.aqrep.2022.101166
Downloads
Published
Issue
Section
License
Copyright (c) 2026 Journal of Academia
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
