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1 J. of Aquaculture Vol. 19(4) : , 2006 µ Journal of Aquaculture Korean Aquaculture Society ü paprika spirulina ƒƒ (Coreoleuciscus splendidus) tv k e w ½, 1, 1, * Ÿ w, 1 w wœw Effect of Dietary Supplementation of Paprika and Spirulina on Pigmentation of Swiri Coreoleuciscus splendidus Suk-Ree Kim, Choong-Ryul Lee 1 and Sang-Min Lee 1, * Busan Institute of Science Education, Busan , Korea 1 Faculty of Marine Bioscience & Technology, Kangnung National University, Gangneung , Korea This study was conducted to evaluate the effect of dietary supplementation of paprika and spirulina as carotenoid sources with three different levels on survival and skin color development of swiri. Fish (initial mean weight 4.4 g) were fed the diets containing different levels of paprika (2, 5 and 10%), spirulina (5, 10 and 20%) and mixture of 5% paprika and 10% spirulina and without supplementation (con) for 9 weeks. Survival of fish fed diets containing the various levels of spirulina was 100% and higher than that of fish fed the other diets. Total carotenoid contents of the skin and fin in fish fed the diets containing the various levels of paprika and spirulina were higher than those in fish fed the control diet. Total carotenoid contents of skin and fin in fish fed the diets containing 5% and 10% paprika tended to be higher than that in fish fed the diet containing 2% paprika. Total carotenoid contents of fish fed the diets containing 10% and 20% spirulina tended to be higher than that of fed the diet containing 5% spirulina. The apparent color definition of skin and fin in fish fed the diets containing paprika and spirulina was better than fish fed the control diet. The results of this study suggest that dietary supplementation with paprika and spirulina can improve skin coloration of swiri. Keywords: Pigmentation, Swiri Coreoleuciscus splendidus, Dietary carotenoid, Skin color ù š û, ve ûe, w w y û wš. w, ü y w w sw ƒ, š j» w v w. w w k w. ü w ƒƒeƒ w, ƒ rw d w w q. ü w w 200, xk, y pw *Corresponding author: smlee@kangnung.ac.kr yƒ w w ƒ ƒ. w, û y ywš p w š. Mori (1935) w» w p w ûw w d w t v (Kim and Park, 2002). tv w carotenoids, melanin, flavine purine. w y carotenoids sw, w w x w ƒ š. ù sww carotenoids ww mw l œ w(guillaume et al., 2001). t 261

2 262 ½,,» w carotenoids ww ƒw w» w ùpv(boonyaratpalin and Unprasert, 1989), (Ha et al., 1997; Baker et al., 2002), (Booth et al., 2004; Kalinowski et al., 2005) w š. w, ü û (Kim et al., 1999a; Kim et al., 1999b) carotenoids w ƒ š. ù ü ü ƒ w z w š. w w ƒw šƒ û ü j ƒ. ü w ƒeƒ w p ü paprika spirulina ƒ ƒƒ ƒ w tv k e w w. x Table 1 tw w w 48%, w w 7% zƒ x w. ƒw, paprika ƒ(2%, 5% 10%), spirulina ƒ(5%, 10% 20%) paprika (5%) spirulina (10%) Table 1. Ingredients and chemical composition of the experimental diets yw ƒw 8 x w. x yww z, 100 g 40 g ƒw x q xw z þ(-25 o C) w œ w. carotenoids w û ùkû, w ƒw ƒ w ƒw carotenoids w ƒw. x q w Ÿ w w. ³» w» w 3% w z, x 2 w.» 1 2z œ w. x s³ 4.4 g z x w 8 x( cm ƒ ) ƒƒ 15 w z, x œ w 9 w. œ 1 2z(10:00, 17:00) œ w. ü 1 1z(20% y) siphonw w. w wš,»» w 17±1 o C ü w. x 30 d w, x ƒ x 3 w ƒ sƒ w le w GGGGGDiets Con P1 P2 P3 S1 S2 S3 P2+S2 IngredientsG(%) White fish meal Soybean meal Wheat flour Squid liver oil Soya oil Vitamin premix Mineral premix Cellulose Paprika powder Spirulina powder Chemical analysis (% of dry matter basis) Crude protein Crude lipid Crude ash Total carotenoids (mg/100g) Vitamin mix. contained the following diluted in cellulose (g/kg mix): ascorbic acid, 92.7; α-tocopheryl acetate, 14.5; thiamin, 2.1; riboflavin, 7.0; pyridoxine, 1.4; niacin, 27.8; Ca-D-pantothenate, 9.7; myo-inositol, 139.1; D-biotin, 4.2; folic acid, 0.5; p-amino benzoic acid, 13.9; K3, 1.4; A, 0.6; D 3, 0.002; choline chloride, 278.3; cyanocobalamin, Mineral mix. contained the following ingreients (g/kg mix): MgSO 4 7H 2O, 80; NaH 2PO 4 2H 2O, 370; KCl, 130; Ferric citrate, 40; ZnSO 4 7H 2O, 20; Ca-lactate, 356.5; CuCl, 0.2; AlCl 3 6H 2O, 0.15; KI, 0.15; Na 2Se 2O 3, 0.01; MnSO 4 H 2O, 2; CoCl 2 6H 2O, 1.

3 w. w, carotenoids w w tv w þ(-75 o C) w. x AOAC (1990) (N 6.25) Auto Kjeldahl System (Buchi B-324/435/ 412, Switzerland; Metrohm 8.719/806, Switzerland) w wš, ether, 105 o C dry oven 6 z dw. z 550 o C zy 4 zy z dw. Carotenoids carotenoids Kim et al., (2003) w w, þ w w. tv carotenoids (MeOH:EtOAc:Petroleum ether = 1:1:1 v/v/v) w 60 sonicationw w, carotenoids yw w z, diethyl ether resuspension g. z 3 ml saturated KOH solution w 16 saponification k z, diethyl ether w carotenoids diethyl ether k z, diethyl ether d yùp(na 2SO 4) kjš, 40 o C w w. 3 ml MeOH:Methyl tert-butyl Ether (1:1 v/v) yw UV-spectrometer w. carotenoids 450 nm Ÿ d w z, t w w. t(standard curve) astaxanthin (Extrasynthese, France) 10 mg 200 ml MeOH:Chloroform (1:1 v/v) yw ww MeOH w ƒ d w. š 9 x z, (Fig. 1) spirulinaƒ ƒ x S1, S2, S3 P2+S2 100% (80%) paprika ƒ (73~93%) ùkû. w, paprika ƒ ü paprika ƒw 2%, 5% 10% ƒw ƒƒ 93%, 80% 73% û. paprika ƒ Fig. 1. Survival of swiri fed the experimental diets for 9 weeks. ü paprika spirulina ƒ tv k 263 Fig. 2. Total carotenoids in the integument of swiri after feeding the experimental diets for 9 weeks. û y ³ w, paprika spirulina w y ƒ w. Spirulina û w vjm w š, k, v γ-linolenic acid ww peptidoglycan s y ƒ(becker and Venkataraman., 1984; Ra, 1991; Miihling and Harris, 2003). Paprika capsanthin capsorubin carotenoid w wš ù s cellulose y û q(park et al., 2005). x z tv carotenoids w (Fig. 2) ƒ 1.58 mg/100 g ƒ û, spirulina 10% ƒ S2 x 2.23 mg/100 g ƒ. Paprika spirulina ƒ w paprika 5% spirulina 10% ƒ tv carotenoids w ƒw. Paprika spirulinaƒ yw ƒ x P2+S mg/100 g paprika spirulinaƒ ƒƒ ƒ P2 (2.14 mg/100 g) S2 (2.23 mg/100 g) û carotenoids w. Choubert and Storebakken (1989) w ü carotenoids w ƒw ü ƒwù» û ƒs û šw. Ha et al. (1997) tv canthaxanthin ƒ ƒ, lutein ƒ ƒ k zƒ š šw. w, w tv astaxanthin monoester ƒw, lutein ƒw k zƒ š š. Kalinowski et al. (2005) tv k w shrimp meal ü astaxanthin zƒ š šw. w, Kim et al. (1999b) eû carotenoids ü astaxanthin ƒ ƒ carotenoids k ƒ zƒ š šw. w l x ù, x

4 264 ½,, ü carotenoids yw. tv carotenoids w P2 x ƒ P3, S2 x ƒ S3 w Choubert and Storebakken (1989) w. w, P2+S2 x ƒ P2 S2 ƒƒ x û carotenoids w ü carotenoids w» ƒ. w œ carotenoids ü y, y p carotenoid. Kim et al. (1999b) eû carotenoids ü x carotenoidsƒ zeaxanthin w zeaxanthin peroxide š šw. Katsuyama and Matsuno (1988) p v œ w canthaxanthin β-carotene, astaxanthin zeaxanthin y, w Kang and Ha (1994) astaxanthin tunaxanthin š w. w mw, x ww» w carotenoids ƒ w w q. w ü paprika spirulina ƒw x tv w yw yw (Fig. 3). p, paprika ƒ P3 ƒ w ùkû, spirulina ƒw x x w yw yw. spirulina w carotenoids phycocyanin chlorophyll w y q(ciferri, 1983). x x» ƒ y ƒ ƒ» z ƒ w. Choubert and Storebakken (1989) ü w ü carotenoidsƒ yš w û yƒ š šw. ü»»y» z l x w ùkù. ü w vƒ.,, pv, šƒ œ w ƒƒ d ww q q» carotenoids w w œ w j w. w, w, w j w ƒw. carotenoids» w tš. Amar et al. (2004) carotenoids ww Dunaliella salina Phaffia rhodozyma ü ƒw œ w zƒ š šw, Waagbo et al. (2003) ü astaxanthin ƒw ü û š w. Tiger shrimp Supamattaya et al. (2005) ü Dunaliella ƒw ƒw white spot syndrome virus (WSSV) stress w w ƒw š w, Chien et al. (2003) Fig. 3. Body color of swiri fed the experimental diets for 9 weeks. A: control, B: P1, C: P2, D: P3, E: S1, F: S2, G: S3, H: P2+S2.

5 ü astaxanthin ƒƒ w stress w w z šw. w, Hu et al. (2006) hybrid tilapia vitamin A β-carotene vitamin A y šw. carotenoids w y» ƒ. carotenoids w» w šw w ƒ w ƒ w. ü w k w x spirulina ƒƒ paprika ƒ, ü ƒw paprika 5%, spirulina 10%ƒ q. (Coreoleuciscus splendidus) ü paprika spirulina ƒ ƒƒ ƒ w k e w w. x s³ 4.4 g w, 8 (Control, P1, P2, P3, S1, S2, S3 P2+S2) x w. P1, P2 P3 paprika ƒƒ 2%, 5% 10% ƒw, S1, S2 S3 spirulina ƒƒ 5%, 10% 20% ƒw w. w P2+S2 paprika 5% spirulina 10% yww w. x z, spirulinaƒ ƒ x S1, S2, S3 P2+S2 100% (80%) paprika ƒ (73~93%) ùkû. tv carotenoids w ƒ 1.58 mg/ 100 g ƒ û, spirulina 10% ƒ S2 x 2.23 mg/100 g ƒ. Paprika spirulinaƒ yw ƒ x P2+S mg/100 g paprika spirulinaƒ ƒƒ ƒ P2 (2.14 mg/100 g) S2 (2.23 mg/100 g) û carotenoids w. tv w ƒ x w y w. šw, ü paprika ƒ 5%, spirulina ƒ 10%ƒ k q. 2005»x(RTI ) w.. šx ü paprika spirulina ƒ tv k 265 Amar, E. C., V. Kiron, S. Satoh and T. Watanabe, Enhancement of innate immunity in rainbow trout (Oncorhynchus mykiss Walbaum) associated with dietary intake of carotenoids from natural products. Fish & Shellfish Immunology, 16, AOAC, Official Methods of Analysis. 15th edition. Association of Official Analytical Chemists. Arlington, Virginia pp. Becker, E. W. and L. V. Venkataraman, Production and utilization of the blue green alga Spirulina in India. Biomass, 4, Baker, R. T. M., A.-M. Pfeiffer, F.-J. Schoner and L.-S. Lemmon, Pigmenting efficacy in fresh-water reared Atlantic salmon, Salmo salar. Animal Feed Sci. Tech., 99, Boonyaratpalin, M. and N. Unprasert, Effects of pigments from different sources on colour changes and growth of red Oreochromis niloticus. Aquaculture, 79, Booth, M. A., R. J. Warner-Smith, G. L. Allan and B. D. Glencross, Effects of dietary astaxanthin source and light manipulation on the skin colour of Australian snapper Pagrus auratus (Bloch & Schneider, 1801). Aquacult. Res., 35, Chien, Y.-H., C.-H. Pan and B. Hunter, The resistance to physical stresses by Penaeus monodon Juveniles fed diets supplemented with astaxanthin. Aquaculture, 216, Choubert, G. and T. Storebakken, Dose response to astaxanthin and canthaxanthin pigmentation of rainbow trout fed various dietary carotenoid concentrations. Aquaculture, 81, Ciferri, O., Spirulina, the edible microorganism. Microbiol Rev., 47, Guillaume, J., S. Kaushik, P. Bergot and R. Metailler, Nutrition and feeding of fish and crustaceans. Springer Verlag, pp Ha, B.-S., M.-J. Kweon, M.-Y. Park, S.-H. Baek, S.-Y. Kim, I.-O. Baek and S.-J. Kang, Comparison of dietary carotenoids metabolism and effects to improve the body color of cultured fresh-water fishes and marine fishes. J. Korean Soc. Food Sci. Nutr., 26, Hu, C.-J., S.-M. Chen, C.-H. Pan and C.-H. Huang, Effects of dietary vitamin A or β-carotene concentrations on growth of juvenile hybrid tilapia, Oreochromis niloticus O. aureus. Aquaculture, 253, Kalinowski, C. T., L. E. Robaina, H. Fernandez-Palacios, D. Schuchardt and M. S. Izquierdo, Effect of different carotenoid sources and their dietary levels on red porgy (Pagrus pagrus) growth and skin colour. Aquaculture, 244, Kang, D.-S. and B.-S. Ha, Metabolism of dietary carotenoids and effects to intensify the body color of cultured sea bass. Bull. Korean Fish. Soc., 27, Katsuyama, M. and T. Matsuno, Carotenoid and vitamin A, and metabolism of carotenoids. astaxanthin, zeaxanthin lutein and tunaxanthin in tilapia Tilapia nilotica. Comp. Biochem. Physiol., 86B, 1 5. Kim, H.-S., Y.-h. Kim, S.-H. Cho and J.-Y. Jo, 1999a. Effects of dietary carotenoids on the nuptial color of the Bitterling (Rhodeus uyekii). J. Korean Fish. Soc., 32, Kim, H.-Y., S.-H. Baek, S.-Y. Kim, K.-I. Geong, M.-J. Kweon, J.- H. Kim and B.-S. Ha, 1999b. Metabolism of dietary carotenoids and effects to improve the body color of Oily Bittering,

6 266 ½,, Acheilognathus koreensis. J. Korean soc. Food Sci. Nutr., 28, Kim, I.-S and J.-Y. Park, Freshwater fishes of Korea. Published by Kyo-Hak Publishing Co., Seoul, Korea, pp Kim, J.-B., S.-H. Ha, J.-Y. Lee, H.-H. Kim, S.-H. Yoon and Y.-H. Kim, Biological activities and analysis of carotenoids in plants. Korean J. Crop Sci., 48(S), Miihling, M. and N. Harris, Reversal of helix orientation in the cyanobacterium Arthrospira. J. Phycol., 39, Mori, T., Studies on the geographical distribution of freshwater fish in Korea. Bull. Biogeogr. Soc. Jap., 7, Park, J.-H., C.-S. Kim and S.-K. Noh, The effect of fresh paprika and paprika powder dried by far-infrared ray on inhibition of lipid oxidation in Lard Model System. Korean J. Food Cookery Sci., 21, Ra, K., Microalgae as food and supplement. Crit Rev. Food Sci. Nutr., 30, Supamattaya, K., S. Kiriratnikom, M. Boonyaratpalin and L. Borowitzka, Effect of a Dunaliella extract on growth performance, health condition, immune response and disease resistance in black tiger shrimp (Penaeus monodon). Aquaculture, 248, Waagbo, R., K. Hamre, E. Bjerkas, R. Berge, E. Wathne, O. Lie and B. Torstensen, Cataract formation in Atlantic salmon, Salmo salar L., smolt relative to dietary pro- and antioxidants and lipid level. J. Fish Diseases, 26, š : :