Effects of lysophospholipid in diet containing astaxanthin on immune indices, antioxidant enzymes, and carotenoid content in koi carp (Cyprinus carpio var. koi)

Document Type : Research Paper

Authors

Department of Fisheries, Faculty of Animal Science and Fisheries, Sari Agricultural Sciences and Natural Resources University, Sari, Mazandaran, Iran

10.22124/janb.2026.33458.1306

Abstract

Introduction: Ornamental fish are aquatic animals primarily reared in aquariums and decorative ponds for aesthetic purposes rather than human consumption. Among freshwater ornamental species, koi carp (Cyprinus carpio var. koi) is one of the most popular due to its remarkable diversity of colors and patterns, which significantly determine its market value. Therefore, understanding pigments and their dietary sources is essential in ornamental aquaculture. Carotenoids are the main compounds responsible for coloration in ornamental fish, however, fish species such as koi cannot synthesize them independently and must obtain them through feed. Astaxanthin, an important carotenoid with strong antioxidant properties, supports health, immunity, and skin coloration. Lysophospholipids may enhance fat-soluble compound absorption, improving astaxanthin uptake. This study evaluated their effects on pigment absorption, immunity, and antioxidant indices in koi carp.
Materials and Methods: A total of 150 koi carp (with an initial mean weight 3.63 ± 0.05 g) were purchased from a local farm and transferred to a private rearing facility. After a two-week acclimation period on a commercial ornamental fish diet, the fish were randomly distributed into 15 glass aquaria (60 L; 60 × 45 × 20 cm) with 5 treatments, 3 replicates, and 10 fish per aquarium. Water quality was maintained by filtration, at temperature 26 ± 1°C, pH 7.5 ± 0.5, and dissolved oxygen 7 ± 0.5 mg/L. Synthetic astaxanthin (>98% purity) and lecithin-based lysophospholipid were added to the basal diet by spraying with canola oil. Treatments included: control, 0.01% astaxanthin, 0.02% astaxanthin, 0.01% astaxanthin + 0.1% lysophospholipid, and 0.02% astaxanthin + 0.1% lysophospholipid. Fish were fed at 5% body weight, three times daily, for 56 days. At the end of the feeding trial, serum lysozyme, alternative complement activity (ACH50), immunoglobulin, and antioxidant enzymes including superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) were measured using standard spectrophotometric methods. Serum, skin, and fillet carotenoids were also assessed. Data were analyzed using One-Way ANOVA followed by Duncan’s test in SPSS version 19, with significance set at P<0.05.
Results: According to the immune parameters, lysozyme activity was significantly lower in the control and diet containing 0.01% astaxanthin compared to other groups (P<0.05). Immunoglobulin and complement activities were also significantly influenced by the different diets, with the combined astaxanthin–lysophospholipid groups showing the highest values (P<0.05). The highest CAT activity was recorded in fish fed the diet containing 0.01% astaxanthin supplemented with 0.1% lysophospholipid (P<0.05). The lowest SOD activity was observed in the control and 0.01% astaxanthin groups, while the addition of lysophospholipid to diets containing astaxanthin significantly increased SOD activity (P<0.05). Likewise, fish receiving the combination of lysophospholipid and astaxanthin, as well as the diet containing 0.02% astaxanthin, showed higher GPx activity than the control group (P<0.05). Carotenoid accumulation in the skin and fillet was significantly higher in fish fed astaxanthin-supplemented diets, particularly when combined to lysophospholipid (P<0.05).
Discussion: Assessment of immune parameters in fish is an important indicator of health and immune efficiency, since these markers reflect both innate and acquired defense capacity. In the present study, adding astaxanthin to lysophospholipid synergistically improved lysozyme, complement, and immunoglobulin activities. This effect was probably due to better astaxanthin absorption caused by lysophospholipid, which enhances bioavailability through mixed micelle formation and facilitation of lipid transport. Antioxidant enzymes such as SOD, CAT, and GPx play crucial roles in mitigating oxidative stress and maintaining fish health. Here, astaxanthin-containing diets increased antioxidant enzyme activities compared to the control, and the best antioxidant performance was observed with combination of 0.01% astaxanthin and 0.1% lysophospholipid. This suggests a synergistic effect between the two additives. Similar improvements in antioxidant capacity have been reported in trout, tilapia, shrimp, and other aquatic species. Astaxanthin also increased carotenoid deposition in koi skin and fillet, improving pigmentation and likely market value. The inclusion of lysophospholipid further enhanced carotenoid accumulation in tissues, probably by improving emulsification and intestinal uptake of this fat-soluble pigment. In contrast, lower serum carotenoid levels in combined treatments may indicate faster transfer from blood to target tissues.
Conclusion: Based on the present research, it can be stated that the combination of lysophospholipid and astaxanthin, particularly at the 0.01% level, likely enhanced tissue deposition of astaxanthin, which may reflect improved bioavailability, and concurrently improved antioxidant capacity and immune function.

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References

Barbosa, M., Morais, R., Choubert, G. 1999. Effect of carotenoid source and dietary lipid content on blood astaxanthin concentration in rainbow trout (Oncorhynchus mykiss). Aquaculture 176: 331-341.
 
Besharat, M., Islami, H.R., Soltani, M., Mousavi, S.A. 2024. Effects of dietary nanoliposome-coated astaxanthin on haematological parameters, immune responses and the antioxidant status of rainbow trout (Oncorhynchus mykiss). Veterinary Medicine and Science 10: 1461. DOI: 10.1002/vms3.1461
 
Bjerkeng, B., Følling, M., Lagocki, S., Storebakken, T., Olli, J.J., Alsted, N. 1997. Bioavailability of all-E-astaxanthin and Z-isomers of astaxanthin in rainbow trout (Oncorhynchus mykiss). Aquaculture 157: 63-82. DOI: 10.1016/S0044-8486(97)00146-4
 
Chen, H.C., Chang, W.C., Chuang, J.Y., Chang, K.Y., Liou, J.P., Hsu, T.I. 2023. The complex role of eicosanoids in the brain: Implications for brain tumor development and therapeutic opportunities. Biochimica et Biophysica Acta (BBA)-Reviews on Cancer 1878: 188957. DOI: 10.1016/j.bbcan.2023.188957
 
Choubert, G., Storebakken, T. 1989. Dose response to astaxanthin and canthaxanthin pigmentation of rainbow trout fed various dietary carotenoid concentrations. Aquaculture 81: 69-77. DOI: 10.1016/0044-8486(89)90231-7
 
Esteban, M.Á., Cuesta, A., Chaves-Pozo, E., Meseguer, J. 2015. Phagocytosis in teleosts. Implications of the new cells involved. Biology 4: 907-922. DOI: 10.3390/biology4040907
 
Huang, Y., Xu, J., Sheng, Z., Chen, N., Li, S. 2021. Integrated response of growth performance, fatty acid composition, antioxidant responses and lipid metabolism to dietary phospholipids in hybrid grouper (Epinephelus fuscoguttatus× E. lanceolatus) larvae. Aquaculture 541: 736728. DOI:  10.1016/j.aquaculture.2020.735813
 
Ibarz, A., Sánchez-Lozano, G., Bamba, J., Pérez-Sánchez, J., Kaushik, S. 2023. Physiological benefits of dietary lysophospholipid supplementation in a marine fish model: deep analyses of modes of action. Animals 13: 1381. DOI: 10.3390/ani13081381
 
Lalita, A., Lili, W., Pratiwy, F.M. 2022. The effect of differences in the addition of astaxanthin and several sources of natural ß-carotene in increasing color intensity of koi fish (Cyprinus carpio L.). Asian Journal of Fisheries and Aquatic Research 18: 38-47. DOI: 10.9734/ajfar/2022/v18i230438
 
Lawrence, R.A., Burk, R.F. 1976. Glutathione peroxidase activity in selenium-deficient rat liver. Biochemical and Biophysical Research Communications 71: 952-958. DOI: 10.1016/0006-291X(76)90728-6
 
Liu, Y., Wang, J., Chen, Q., Li, X. 2024. Dual-function analysis of astaxanthin on golden pompano (Trachinotus ovatus): Growth performance and antioxidative regulation. Aquaculture Research 55: 3489-3503. DOI: 10.1002/aqr.5678
 
Liu, Y., Chen, G., Li, X., Xu, C., Zhou, Q. 2018. Lysophospholipid enhances intestinal health and disease resistance in aquatic animals. Fish and Shellfish Immunology 77: 113-120. DOI: 10.1016/j.fsi.2018.03.039
 
Mahasri, G., Widyastuti, P., Sulmartiwi, L. 2011. Leukocyte profile of koi fish (Cyprinus carpio) which infested by Ichthyophthirius multifiliis on the different infestation degree with cohabitation methode. Jurnal Ilmiah Perikanan dan Kelautan 3: 91. DOI: 10.20473/jipk.v3i1.11629
 
Marklund, S., Marklund, G. 1974. Involvement of the superoxide anion radical in the autoxidation of pyrogallol and a convenient assay for superoxide dismutase. European Journal of Biochemistry 47: 469-474. DOI: 10.1111/j.1432-1033.1974.tb03714.x
 
Melo, N., de Souza, S.P., Konig, I., de Jesus Paula, D.A., Ferreira, I.S., Luz, R.K., Murgas, L.D.S. 2024. Sensitivity of different organs and tissues as biomarkers of oxidative stress in juvenile tambaqui (Colossoma macropomum) submitted to fasting. Comparative Biochemistry and Physiology Part A: Molecular & Integrative Physiology 291: 111595. DOI: 10.1016/j.cbpa.2024.111595
 
Mettelman, R.C., Souquette, A., Van de Velde, L.A., Vegesana, K., Allen, E.K., Kackos, C.M., Trifkovic, S., DeBeauchamp, J., Wilson, T.L., St. James, D.G., Menon, S.S. 2023. Baseline innate and T cell populations are correlates of protection against symptomatic influenza virus infection independent of serology. Nature Immunology 24: 1511-1526. DOI: 10.1038/s41590-023-01550-0
 
Omar, E., Chiodi, C. 2024. Fat digestion and absorption: Normal physiology and pathophysiology of malabsorption, including diagnostic testing. Nutrition in Clinical Practice 39: S6-S16. DOI: 10.1002/ncp.10841
 
Pei, J., Liu, W., Han, L. 2019. Research on evaluation index system of Chinese city safety resilience based on Delphi method and cloud model. International Journal of Environmental Research and Public Health 16: 3802. DOI: https://doi.org/10.3390/ijerph16203802
 
Perry, W.B. 2023. The environmental impact of keeping a tropical aquarium in Northern Europe. Journal of Fish Biology 103: 695-703. DOI: 10.1111/jfb.15258
 
Rahmati, R., Nayebi, B., Ayati, B. 2021. Investigating the effect of hydrogen peroxide as an electron acceptor in increasing the capability of slurry photocatalytic process in dye removal. Water Science and Technology 83: 2414-2423. DOI: 10.1111/jfb.15258
 
Sallam, A.E., Kotit, A.M., Almisherfi, H.M. 2024. Dietary lysophospholipid improves growth performance, antioxidant capacity and immunity response of red tilapia (Oreochromis niloticus× O. Mossambicus). Egyptian Journal of Aquatic Research 50: 424-429. DOI: 10.1016/j.ejar.2024.04.002
 
Sen, S., Sirobhushanam, S., Johnson, S.R., Song, Y., Tefft, R., Gatto, C., Wilkinson, B.J. 2016. Growth-environment dependent modulation of Staphylococcus aureus branched-chain to straight-chain fatty acid ratio and incorporation of unsaturated fatty acids. PloS One 11: e0165300. DOI: 10.1371/journal.pone.0165300
 
Siwicki, A.I. 1993. Nonspecific defense mechanisms assay in fish. II. Potential killing activity of neutrophils and monocytes, lysozyme activity in serum and organs and total immunoglobulin (Ig) level in serum. Fish Diseases Diagnosis and Prevention Methods. 105-111.
 
Sletvold, N., Trunschke, J., Smit, M., Verbeek, J., Ågren, J. 2016. Strong pollinator-mediated selection for increased flower brightness and contrast in a deceptive orchid. Evolution 70: 716-724. DOI: 10.1111/evo.12881
 
Song, H., Liang, Y., Yang, Y., Liu, C., Liu, Y., Mu, X., Wang, X. 2025. Dietary supplementation with whole-fat or defatted Antarctic krill powder improves the growth performance, body coloration, and immune capability of red–white koi carp (Cyprinus carpio var. koi). Animals 15: 1561. DOI: 10.3390/ani15111561
 
Sukhovskaya, I.V., Lysenko, L.A., Fokina, N.N., Kantserova, N.P., Borvinskaya, E.V. 2023. Survival, growth performance, and hepatic antioxidant and lipid profiles in infected rainbow trout (Oncorhynchus mykiss) fed a diet supplemented with dihydroquercetin and arabinogalactan. Animals 13: 1345. DOI: 10.3390/ani13081345.
 
Taghavizadeh, M., Shekarabi, S.P.H., Mehrgan, M.S., Islami, H.R. 2020. Efficacy of dietary lysophospholipids (Lipidol™) on growth performance, serum immuno-biochemical parameters, and the expression of immune and antioxidant-related genes in rainbow trout (Oncorhynchus mykiss). Aquaculture 525: 1-11. DOI: 10.1016/j.aquaculture.2020.735315
 
Tan, K., Zhang, H., Zheng, H. 2024. Carotenoid content and composition: A special focus on commercially important fish and shellfish. Critical Reviews in Food Science and Nutrition 64: 544-561. DOI: 10.1080/10408398.2022.2101889.
 
Torrissen, O.J. 1989. Pigmentation of salmonids: Interactions of astaxanthin and canthaxanthin on pigment deposition in rainbow trout. Aquaculture 79: 363-374. DOI: .1016/0044-8486(89)90478-X
 
Tripathi, G., Dubey, P., Ahmad, S., Farooqui, A., Mishra, V. 2024. Role of algal-derived bioactive compounds in human health. Recent Patents on Biotechnology 18: 190-209. DOI: 10.2174/1872208317666230623141740
 
Weber, D., Grune, T. 2023. Carotenoid basics: from food to skin. Redox Experimental Medicine DOI: 10.20517/rem.2023.03
 
Yan, H., Wang, Y., Liang, H., Duan, Y., Wang, J., Zhou, C., Huang, Z. 2025. Effects of lysophospholipids on the antioxidant capacity, digestive performance, and intestinal microbiota of Litopenaeus vannamei. Biology 14: 90. DOI:  10.3390/biology14010090.
 
Yano, T. 1992. Assays of hemolytic complement activity. Techniques in fish Immunology 131-141.
 
Ytrestøyl, T., Bjerkeng, B. 2007. Dose response in uptake and deposition of intraperitoneally administered astaxanthin in Atlantic salmon (Salmo salar L.) and Atlantic cod (Gadus morhua L.). Aquaculture 263: 179-191. DOI: 10.1016/j.aquaculture.2006.10.021
 
Zhang, W., Dan, Z., Zhuang, Y., Zheng, J., Gong, Y., Liu, Y., Mai, K., Ai, Q. 2022. Effects of dietary lipid levels on growth, digestive enzyme activities, antioxidant capacity, and lipid metabolism in turbot (Scophthalmus maximus L.) at three different stages. Aquaculture Nutrition 104226. DOI:  10.1155/2022/104226.

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