Dietary Supplementation of Ulva sp. Water Extract Improves Growth and Resistance to Hypoxia Stress in Zebrafish (Danio rerio)
DOI:
https://doi.org/10.61741/JMBI.2026.v4.p%25pKeywords:
hypoxia stress, polysaccharides, supplementation, Ulva sp., zebrafishAbstract
Dissolved oxygen depletion in intensive aquaculture systems poses a threat to fish immunity and survival. Ulva sp. contains bioactive polysaccharides, particularly ulvan, which have demonstrated antioxidant and immunostimulatory properties. This study investigated the effects of Ulva sp. water extract supplementation at various doses on growth performance and hypoxia stress resistance in zebrafish (Danio rerio). Before feeding trials, toxicity was assessed using Artemia sp. bioassay at concentrations of 125,000; 62,500; 31,250; 15,625; and 7,813 ppm to determine the LC50 value. A completely randomized design (CRD) consisting of five dietary treatments (0%, 2.5%, 5%, 10%, and 20% of the LC₅₀ value) with three replicates per treatment was employed. Fish were fed the experimental diets twice daily for 30 days and subsequently subjected to hypoxia stress tests in sealed airtight aquaria. The LC₅₀ value of Ulva sp. water extract was 44,977.92 ppm, indicating non-toxicity. Dietary supplementation significantly enhanced growth performance, with the highest absolute weight gain recorded in fish receiving 20% of the LC₅₀ supplementation level (167 ± 0.02 mg; p < 0.05). Supplementation also improved resistance to hypoxia stress, as evidenced by significantly higher survival rates in the 10% and 20% LC₅₀ treatments (57%) compared with the control group (p < 0.05). However, survival during the 30-day feeding period did not differ significantly among treatments (p > 0.05). Overall, the results suggest the potential of Ulva sp. water extract as a functional feed additive, while further research is required to determine optimal supplementation levels and feeding duration for enhanced aquaculture performance.
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Adhish, M., and Manjubala, I. 2023. Effectiveness of zebrafish models in understanding human diseases—A review of models. Heliyon, 9(3). https://doi.org/10.1016/j/heliyon.2023.314557.
Akbary, P., and Z. Aminikhoei. 2018. Effect of polysaccharides extracts of algae Ulva rigida on growth, antioxidant, immune response and resistance of shrimp, Litopenaeus vannamei against Photobacterium damselae. Aquaculture Research 49(2): 1-8. https://doi.org/10.1111/are.13710.
Aqiila, G. R., I. Taufiqurrahman, and E. Wydiamala. 2017. Uji Efektifitas Ekstrak Etanol Daun Ramania (Bouea macrophylla Griffith) terhadap Mortalitas Larva Artemia salina Leach. Jurnal Kedokteran Gigi 2(2): 170-176. http://dx.doi.org/10.20527/dentino.v2i2.3995.
Bourguiba, I., A. Zahlila, N. Bouaicha, M. Amri, and S. Mezghani. 2017. Antioxidant effect of the marine green alga Ulva rigida ethanolic precipitate in yeast cells and zebrafish embryos. South Africa Journal of Botany 113: 253-260. https://doi.org/10.1016/j.sajb.2017.09.001.
Charoeythornkhajhornchai, P., T. Kunjiek, S. Chaipayang and S. Phosri. 2023. Toxicity assessment of bioplastics on brine shrimp (Artemia franciscana) and cell lines. Emerging Contaminants 9(4): 1-14. https://doi.org/10.1016/j.emcon.2023.100253.
Chen, J., W. Zeng, J. Gan, Y. Li, Y. Pan, J. Li, and H. Chen. 2021. Physicochemical properties and anti-oxidation activities of ulvan from Ulva pertusa Kjellm. Alga Research 55: 1-8. https://doi.org/10.1016/j.algal.2021.102269.
Hariati, Y., A. Thaib, S. Muhazzir, Devri alvandi, and R. Maulidya. 2024. Efektifitas Penggunaan Ekstrak Labu Kuning (Cucurbita moscheta Durch) dalam Pakan untuk Mengingkatkan Pigmen Ikan Mas Koki Lion Head (Carassius auratus). Jurnal Agroristek 7(1): 11-13. https://doi.org/10.47647/jar.v7i1.2309.
Hasan, H., A. M. A. Suryadi, F. Hiola, D. R. P. Papeo, dan I. I. Salwa. 2023. “Uji Toksisitas Ranting Patah Tulang (Euphorbia tirucalli L.) Menggunakan Metode Brine Shrimp Lethality Test (BSLT).” Journal Syifa Sciences and Clinical Research 5(3): 382-391. https://doi.org/10.37311/jsscr.v5i3.23237.
Khan, F. R., dan S. S. Alhewairini. 2018. Zebrafish (Danio rerio) as a Model Organism. Current Trends in Cancer Managemen, November 27, 1-12.
Lawrence, C., J. B. A. James, and K. Maloney. 2012. The effects of feeding frequency on growth and reproduction in zebrafish (Danio rerio). Aquaculture 368-369: 103-108. https://doi.org/10.1016/j.aquaculture.2012.09.022.
Lestari, D., R. Kartika, and E. Marliana. 2019. Uji Brine Shrimp Lethality Test (BSLT) Umbi Bawang Tiwai (Eleutherine bulbosa (Mill.) Urb) dan Uji Toksisitas Akut Fraksi Aktif. Jurnal Riset Kefarmasian Indonesia 1(1): 1-10. https://doi.org/10.33759/jrki.v1i1.43.
Mayasari, R. 2017. Pengaruh Limbah Cair Tahu terhadap Mortalitas dan Histopatologi Ginjal Ikan Mas (Cyprinus carpio) sebagai Alternatif Materi Biologi SMA Kelas X. Jurnal Pendidikan Biologi Indonesia 3(2): 123-132. https://doi.org/10.22219/jpbi.v3i2.3907.
Meyer, B. N., N. R. Ferrigni, J. E. Putnam, L. B. Jacobsen, D. E. Nichols, and J. L. McLaughlin. 1982. Brine Shrimp: A Convenient General Bioassay for Active Plant Constituents. Planta Medica 45(5): 31-34. https://doi.org/10.1055/s-2007-971236.
Mohan, K., A. M. Padmanaban, V. Uthayakumar, R. Chandirasekar, T. Muralisankar, and P. Santhanam. 2016. Effect of dietary Ganoderma lucidum polysaccharides on biological and physiological responses of the giant freshwater prawn Macrobrachium rosenbergii. Aquaculture 46: 42-49. https://doi.org/10.1016/j.aquaculture.2016.05.046.
Olmeda, J.F. dan F. J. S. Vazquez. 2011. Thermal biology of zebrafish (Danio rerio). Journal of Thermal Biology 36(2): 91-104. https://doi.org/10.1016/j.jtherbio.2010.12.005.
Pappou, S., Bakopoulos, V., Valsamidis, M. A., Krokida, M., and Batjakas, I. 2024. Supplementation of a commercial diet of European seabass by an algal ethanolic extract of Ulva lactuca. Bull. Univ. Agric. Sci. Vet. Med. Cluj-Napoca Food Sci. Technol, 80, 157-162. https://doi.org/10.15835/buasvmcn-fst:2023.0026.
Pawestri, S., & Arsyi, E. K. 2025. Kajian Pustaka: Antioksidan Alami dari Ulva spp. dan Implikasinya terhadap Perlindungan Seluler. Jurnal Kolaboratif Sains, 8(9), 5679-5694. https://doi.org/10.56338/jks.v8i9.7920.
Pezeshk, F., S. Babaei, A. A. Kenari, M. Hedayati, and M. Naseri. 2018. The effect of supplementing diets with extracts derived from three different species of macroalgae on growth, thermal stress resistance, antioxidant enzyme activities and skin colour of electric yellow cichlid (Labidochromis caeruleus). Aquaculture Nutrition 5(6): 1-8. https://doi.org/10.1111/anu.12869.
Prama, H., M. Nur, dan E. Ayuzar. Pengaruh penambahan bahan pengencer sperma terhadap fertilitas spermatozoa ikan lele dumbo (Clarias gariepinus). Aquatic Sciences Journal 1(1): 46-52. https://doi.org/10.29103/aa.v1i1.298.
Premarathna, A. D., T. A. E. Ahmed, V. Rjabovs, A. T. Critchley, M. T. Hincke, and R. Tuvikene. 2024. Green seaweed-derived polysaccharides: Insights into various bioactivities for biomedical applications. International Journal of Biological Macromolecules 282(4): 1-17. https://doi.org/10.1016/j.ijbiomac.2024.136858.
Purnamasari, E., E. Purwaningsih, D. Mukhtar, and A. Giantini. 2024. Pengaruh Stress Oksidatif pada Ginjal terhadap Kadar Klotho. Jurnal Penelitian Kedokteran dan kesehatan 6(2): 623-630. https://doi.org/10.31970/ma.v6i3.218.
Qi, H., T. Zhao, Q. Zhang, Z. li, Z. Zhao, and R. Xing. 2005. Antioxidant activity of different molecular weight sulfated polysaccharides from Ulva pertusa Kjellm (Chlorophyta). Journal of Applied Phycology 17(6): 527-534. https://doi.org/10.1007/s10811-005-9003-9.
Qualhato, G., F. C. Dias, and T. L. Rocha. 2024. Hazardous effects of plastic microfibres from facial masks to aquatic animal health: Insights from zebrafish model. Science of the Total Environment 951(369): 1-16. https://doi.org/10.1016/j.scitotenv.2024.175555.
Ramadhan, W., Uju, S. D. Hardiningtyas, R. F. Pari, Nurhayati, and D. Sevica. 2022. Ulvan Polysaccharide Extraction from Ulva sp. Seaweed Assisted by Ultrasonic Waves at Low Temperature. Indonesian Journal of Fishery Product Processing 25(1): 132-142. http://dx.doi.org/10.17844/jphpi.v25i1.40407.
Rama Nisha, P., Elezabeth Mary, A., Uthayasiva, M., and Arularasan, S. 2014. Seaweed Ulva reticulata a potential feed supplement for growth, colouration and disease resistance in fresh water ornamental gold fish, Carassius auratus. J Aquac Res Development, 5(254), 2. https://doi.org/10.4172/2155-9546.1000254.
Setyati, W. A., M. Zainuddin, and P. P. Renta. 2017. Pathogenic Assay of Probiotic Bacteria Producing Proteolytic Enzymes as Bioremediation Bacteria Against Vannamei Shrimp Larvae (Litopenaeus vannamei). Indonesian Journal of Marine Sciences 22(2): 93-98. https://doi.org/10.14710/ik.ijms.22.2.93-98.
Situmorang, M. L., P. Nurwidayanti, and G. Suantika. 2021. Synbiotic Containing Kappaphycus alvarezii, Spirulina sp., dan Halomonas alkaliphila Improves Survival, Growth and Vibriosis Resistance in Whiteleg Shrimp (Litopenaeus vannamei) Post-Larval Culture. Aquatic Living Resources 34(10): 1–8. https://doi.org/10.1051/alr/2021009.
Son, M. A. M., S. Elbahnaswy, M. A. Khormi, A. M. Aborasain, H. H. Abdelhaffez and E. Zahran. 2025. Harnessing the fish gut microbiome and immune system to enhance disease resistance in aquaculture. Fish and Shellfish Immunology 163(1): 1-23. https://doi.org/10.1016/j.fsi.2025.110394.
Souza, A. M., F., F. C. S. Junior, E. D. Dantas, M. C. G. Pereira, S. R. B. Medeiros, and A. C. Luchiari. 2025. Temperature effects on development and lifelong behavior in zebrafish. Science of the Total Environment 973: 1-13. https://doi.org/10.1016/j.scitotenv.2025.179172.
Sunaryo, S., A. O. Saputra, J. R. Hidayati, dan I. E. Susetya. 2024. Extraction of Sulfated Polysaccharides from Ulva sp. Using Acid and Toxicity Testing with the Brine Shrimp Lethality Test (BSLT). Journal of Marine Biotechnology and Immunologi 2(3): 36-41. https://doi.org/10.61741/rtj7qn57.
Suratno, S. and Putra, D. F. 2021. Ectoparasite Control in Sangkuriang Catfish (Clarias sp.) Using Dissolved Oxygen Concentration as a Limiting Factor. Journal of Vocational Fisheries Sciences. 2(2): 32-36. http://dx.doi.org/10.35726/jvip.v2i2.597.
Wang, Y. C., and Y. Chang Chen. 2016. “Extraction and Characterization of Fucoidan from Six Brown Macroalgae.” Journal of Marine Science and Technology 24(2): 319-328. https://doi.org/10.6119/JMST-015-0521-3.
Yousefi, S., S. H. Hoseinifar, H. Paknejad, and A. Hajimoradloo. 2018. The effects of dietary supplement of galactooligosaccharide on innate immunity, immune related genes expression and growth performance in zebrafish (Danio rerio). Fish and Shellfish Immunology 73: 192-196. https://doi.org/10.1016/j.fsi.2017.12.022.
Yudiati, E., Rustadi, F. I. Ginzel, J. R. Hidayati, M. S. Rizfa, N. Azhar, M. S. R. Djarod, E. Heriyati, and R. Alghazeer. 2020. “Oral Administration of Alginate Oligosaccharide from Padina sp. Enhances Tolerance of Oxygen Exposure Stress in Zebrafish (Danio rerio).” Indonesian Journal of Marine Sciences 25(1): 7-14. https://doi.org/10.14710/ik.ijms.25.1.7-14.
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