U. Yanuhar, Z. Waheed, H. Suryanto, V. Hasan, D.R. Machfuda
BACKGROUND AND OBJECTIVES: The cantang grouper (E. fuscoguttatus × E.lanceolatus) is an important marine cultured fish in Indonesia; however, its production has collapsed from approximately 10,208 tons in 2023 to 3,632 tons in 2024. This decrease is mainly attributed to repeated occurrences of viral nervous necrosis, leading to mortality rates approaching 100 percent. The objectives of this study are to investigate whether zinc oxide-chitosan hybrid nanoparticles can serve as an effective nanovaccine delivery system, utilizing recombinant Chlorella vulgaris protein to enhance protective immunity against viral nervous necrosis in cantang grouper. METHODS: An in vivo assessment was conducted using 100 juvenile hybrid groupers, which were divided into five experimental groups: untreated healthy controls, viral nervous necrosis challenged controls, and three vaccinated receiving escalating nanovaccine doses (33 microliters, 66 microliters, and 112 microliters). Each treatment group consisted of 20 fish. Sampling occurred at baseline (day 0) and post-treatment (day 56), with collection of blood and target tissues (kidney) for comprehensive immune and histological assessment via flow cytometry, hematological profiling, and antioxidant enzyme. The methodology of one-way analysis of variance was employed for statistical comparisons. FINDINGS: The lowest tested dose 33 microliters elicited the most pronounced immunoprotective response. Specifically, Interferon gamma levels increased by 11.90 percent, tumor necrosis factor-alpha levels rose by 5.34 percent, and Immunoglobulin M levels increased by 4.75 percent. Additionally, the activities of superoxide dismutase (3.213 unit per milliliter), catalase (109.7 unit per liter), and glutathione peroxidase (76.02 micromol) were significantly elevated, indicating a more robust defense against free radicals. Histopathological examination revealed negligible renal damage in T3, with edema (10.71 percent), necrosis (12.50 percent), and hemorrhage (13.45 percent), compared to more severe damage in the viral nervous necrosis challenged control group (T2). CONCLUSION: Zinc oxide–chitosan hybrid nanoparticles effectively function as nanovaccine, enhancing the immune response and protecting cantang grouper from viral nervous necrosis. The 33 microliters dosage delivered optimal protection, bolstering immune function as well as tissue integrity. This nanovaccine provides a cost-efficient and scalable approach to bolster disease resilience in grouper aquaculture, rendering it a promising instrument for enhancing the sustainability of the industry. © 2026 The author(s). This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third-party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit: http://creativecommons.org/licenses/by/4.0/
Resources Management, Faculty of Fisheries and Marine Sciences, Universitas Brawijaya, Jl. Veteran East Java, Malang, Indonesia; Borneo Marine Research Institute, Universiti Malaysia Sabah, Jalan UMS Sabah, Kota Kinabalu, 88400, Malaysia; Department of Mechanical and Industrial Engineering, Center Excellent for Cellulose Composite (CECCom), Faculty of Engineering, Universitas Negeri Malang, Jl. Semarang 5, Malang, 65145, Indonesia; Study Program of Aquaculture, Faculty of Fisheries and Marine Sciences, Universitas Airlangga, Jl. Mulyorejo, East Java, Surabaya, Indonesia; Master Program in Aquaculture, Faculty of Fisheries Marine Science, Universitas Brawijaya, Jl. Veteran East Java, Malang, Indonesia
