J. of Aquaculture Vol. 19(4) : 261-266, 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 611-089, Korea 1 Faculty of Marine Bioscience & Technology, Kangnung National University, Gangneung 210-702, 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
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(60 40 40 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 41 41 41 43 38 33 23 32 Soybean meal 28 28 28 28 28 28 28 28 Wheat flour 19 18 17 10 18 18 18 15.4 Squid liver oil 2 2 2 2 2.2 2.5 3.3 2.6 Soya oil 2 2 2 2 2 2 2 2 Vitamin premix 1 2 2 2 2 2 2 2 2 Mineral premix 2 3 3 3 3 3 3 3 3 Cellulose 3 2 1.8 1.5 0.7 Paprika powder 2 5 10 5 Spirulina powder 5 10 20 10 Chemical analysis (% of dry matter basis) Crude protein 48.6 48.3 48.1 48.1 49.5 49.1 48.5 47.9 Crude lipid 7.2 7.2 7.2 7.3 7.2 7.1 7.2 7.1 Crude ash 12.5 12.5 12.4 12.7 11.9 11.1 9.4 10.9 Total carotenoids (mg/100g) 0.2 17.0 41.5 56.9 19.5 50.9 132.9 84.0 1 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, 0.003. 2 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.
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+S2 1.61 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
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.
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+S2 1.61 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(RTI05-01-02) w.. šx ü paprika spirulina ƒ tv k 265 Amar, E. C., V. Kiron, S. Satoh and T. Watanabe, 2004. Enhancement of innate immunity in rainbow trout (Oncorhynchus mykiss Walbaum) associated with dietary intake of carotenoids from natural products. Fish & Shellfish Immunology, 16, 527 537. AOAC, 1990. Official Methods of Analysis. 15th edition. Association of Official Analytical Chemists. Arlington, Virginia. 1298 pp. Becker, E. W. and L. V. Venkataraman, 1984. Production and utilization of the blue green alga Spirulina in India. Biomass, 4, 105 125. Baker, R. T. M., A.-M. Pfeiffer, F.-J. Schoner and L.-S. Lemmon, 2002. Pigmenting efficacy in fresh-water reared Atlantic salmon, Salmo salar. Animal Feed Sci. Tech., 99, 97 106. Boonyaratpalin, M. and N. Unprasert, 1989. Effects of pigments from different sources on colour changes and growth of red Oreochromis niloticus. Aquaculture, 79, 375 380. Booth, M. A., R. J. Warner-Smith, G. L. Allan and B. D. Glencross, 2004. Effects of dietary astaxanthin source and light manipulation on the skin colour of Australian snapper Pagrus auratus (Bloch & Schneider, 1801). Aquacult. Res., 35, 458 464. Chien, Y.-H., C.-H. Pan and B. Hunter, 2003. The resistance to physical stresses by Penaeus monodon Juveniles fed diets supplemented with astaxanthin. Aquaculture, 216, 177 191. Choubert, G. and T. Storebakken, 1989. Dose response to astaxanthin and canthaxanthin pigmentation of rainbow trout fed various dietary carotenoid concentrations. Aquaculture, 81, 69 77. Ciferri, O., 1983. Spirulina, the edible microorganism. Microbiol Rev., 47, 551 578. Guillaume, J., S. Kaushik, P. Bergot and R. Metailler, 2001. Nutrition and feeding of fish and crustaceans. Springer Verlag, pp. 183 196. Ha, B.-S., M.-J. Kweon, M.-Y. Park, S.-H. Baek, S.-Y. Kim, I.-O. Baek and S.-J. Kang, 1997. 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, 270 284. Hu, C.-J., S.-M. Chen, C.-H. Pan and C.-H. Huang, 2006. Effects of dietary vitamin A or β-carotene concentrations on growth of juvenile hybrid tilapia, Oreochromis niloticus O. aureus. Aquaculture, 253, 602 607. Kalinowski, C. T., L. E. Robaina, H. Fernandez-Palacios, D. Schuchardt and M. S. Izquierdo, 2005. Effect of different carotenoid sources and their dietary levels on red porgy (Pagrus pagrus) growth and skin colour. Aquaculture, 244, 223 231. Kang, D.-S. and B.-S. Ha, 1994. Metabolism of dietary carotenoids and effects to intensify the body color of cultured sea bass. Bull. Korean Fish. Soc., 27, 272 281. Katsuyama, M. and T. Matsuno, 1988. 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, 276 279. 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,
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