國立中山大學學位論文典藏

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1 (cobia, Rachycentron canadum) ( 1993) (Robins and Ray, 1986) 15 (, 1999) (, 1999) (Lee and Putnam, 1973 ; Reinitz and Hitzel, 1980 ; Gatlin and Stickney, 1982 ; Millikin, 1983) Tucker et al. (1997) % Casledine and Buckely (1980) 4 % 1

2 Takeuchi et al. (1991) 15 % (Murai et al., 1985) (Ellis and Reigh, 1991) gilthead seabream (Vergara et al., 1999) C18:3 EPA DHA (Yamada et al, 1980 Watanabe, 1989 Thongrod et al., 1989 Takeuchi et al., 1990 Sargent et al., 1999) n-3 (Kanazawa et al., 1980) (Yone et al., 1975) n-3 Bruce et al. (1993) n-3 EPA DHA (Furuita et al., 1998) DHA EPA EPA/DHA (Watanabe, 1993 ) (Lee and Putnam,

3 Garling and Wilson, 1977 Bromley, 1980 Millikin, 1983 Hillestrad and Johnsen, 1994) (Cowey et al.,1975) (Hemre and Sandnes,1999) (Ellis and Reigh, 1991) (Watanabe, 1982; Xu et al., 1993) Takeda et al.(1975) 8 % 17% 71% 55% (Lee and Putman, 1973) ( Bromley, 1980) (Borlongan, 1992) (high unsaturated fatty acid; HUFA) 3

4 ( Yu, et al., 1977) 4

5 ( ) ±3.80 g ±2.95g 5 8 FRP ( 1m cm ) ppt 5% ( 67%) Casein( 80.2%) 45% ( ) / (2:1) CMC 24 % 15 4mm ( ) % (L5) 10 % (L10) 15% (L15) 20% (L20) 25% (L25) 5

6 . Table 1. Ingredient composition (%) of the test diets in the experimental Composition L5 L10 L15 L20 L25 Fish meal Casein Alpha starch Corn starch Cod liver oil / Corn oil(2:1) Vitamin mix. (a) Mineral mix. (b) Cholin chloride K 2 HPO Ca(H 2 PO 4 ) CMC (C) a Contained (as g/kg):vitamin A, 600 IU/g; Vitamin D 3,100 IU/g; Vitamin E,50; Vitamin K 3,2.5; Thiamin,10; Riboflavin,15; Pyridoxine,7.5; Cyanocobalamin,0.025; Pantothenate,27.5; Biotin,0.5; Folic Acid, 2.5; Inositol, 100. b Contained(as g/kg):feso 4.7H 2 O,35; MgSO 4.7H 2 0,133; AlCl 3.6H 2 O,7; ZnSO 4.7H 2 0,20; CuSO 4.5H 2 O,10; NaCl,50; KI,0.05. c CMC: Carboxymethyl cellulose. 6

7 . Table 2. Proximate compositions of fish and casein used in the experiments. Protein Sources Crude Protein Crude lipid Moisture Ash Fish meal Casein

8 . Table 3. The suppliers of the dietary ingredients. Ingredients Fish meal Casein Vitamin-free Corn Starch Alpha Starch Canola oil Corn oil Soybean oil DHA Cod liver oil Source Biobio, Chile Sigma, St. Louis, MO. U.S.A. Eridania Co, Germany Taiflower Co, Tailand President Co, Taiwan President Co, Taiwam President Co, Taiwan Martek Biosciences Co,USA Taiaqua Co, Taiwan 8

9 . Table 4. Proximate analysis (mean±sd n=3) of the diets used in the experimental ( g / 100 g diet ) Dietary lipid(%) Crude protein Lipid Moisture Ash ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ±0.1 9

10 ( ) : AOAC ( % ) g 1 + g 2 1 g : AOAC ( % ) ( ) S 1 + ( ) S ( ) 10

11 : AOAC ( 5 KSO HgO ) 15 ml ( Digestion System Digester ) ( 420 ) 75 ml 50 ml ( 50 ) ( Rapidstill, Labconco) 4 % ( Boric acid ) 5 Methyl red Bromocresol green, 0.5 N HCl : ( b - a ) ( % ) = a ml b ml S g Sample ( g ) 10 Folch (1957) (v/v=2:1) 7000rpm 5000rpm 5 (Beckman J2-MC) (v/v=2:1) (%) = ( - )/

12 ( ) (1) percent weight gain (2). food conversion ratio, FCR 1 FCR 100 (3). (protein efficiency ratio, PER) PER= FCR (4) - (hepatosomatic index, HSI) (g) - (%)= 100 (g) 12

13 ( ) 1ml 1.5ml microcentrifuge test tube (Hettich, universal 3RF,Japan)2500rpm 25 (Chemistry analyzer,point Scientific,Inc.,USA) Aspartate transaminase (AST) Alanine transaminase (ALT) Huamn ( ) SAS/PC analysis of variance ( Duncan's new multiple range test (SAS/PC program) 13

14 (weight gain percent) (FCR) (PER) 15% 20% 25% 5% 10% 15% 5% 20% 25% 5% 15% 25% 5 % ( ) 14.6% ( ) 2-4 5% 10% 20% 25% 25% 25% 35.1% 15% 20% 5% 10% 27.1% 14

15 500 Weight gain (%) % 10% 15% 20% 25% Weeks. Fig 1. Percent weight gain of cobia fed diets containing various levels of lipids. 15

16 Table 5. Weight gain, feed conversion rate and protein efficiency ratio of the cobia fed diets containing various levels of lipids 1. Dietary Initial weight Final weight Weight gain FCR PER Lipid, % ( g ) ( g ) ( % ) ± ± 3.80 c ± 1.60 c 1.36 ± 0.01 a 1.63 ± 0.02 b ± ± ab ± ab 1.25 ± 0.07 ab 1.79 ± 0.09 ab ± ± 6.90 a ± 4.90 a 1.17 ± 0.02 b 1.89 ± 0.03 a ± ± 4.90 bc ± 7.40 bc 1.26 ± 0.02 ab 1.76 ± 0.04 ab ± ± 6.30 bc ± bc 1.21 ± 0.03 b 1.83 ± 0.04 a 1 Means (± S.E.) with different superscript letters are significantly different (p<0.05). 16

17 max:14.6. Fig 2. Regression of dietary lipid levels and weight gain of Rachycentron canadum. 17

18 . 8 Table 6. Hepatosomatic index (mean +SD n=3) of the cobia fed diets containing various levels of lipids for 8 weeks. Dietary lipid(%) Hepatosomatic index (%) ± ± ± ± ±

19 % 10% 15% 20% 25% lipid level (%) Weeks Fig3. Lipid contents of ventral muscle of the cobia fed diet containing various levels of lipids.. 19

20 % 10 % 15 % 20 % 25 % lipid level (%) Weeks Fig 4. Lipid contents of dorsal muscle of the cobia fed diets containing various levels of lipids.. 20

21 % 10 % 15 % 20 % 25 % lipid level (%) Weeks Fig 5. Lipid contents of liver of the cobia fed diets containing various levels of lipids.. 21

22 . (%) Table 7. Liver proximate composition ( % ) of the the cobia fed diets containing various levels of lipids for 8 weeks 1. Dietary Lipid(%) Crude Protein Crude Lipid Moisture Ash 5 8.8± ±0.3 c 59.7±0.4 a 1.4± ± ±0.2 c 57.9±0.5 a 1.4± ± ±0.4 b 55.7±0.1 b 1.3± ± ±0.2 b 53.7±0.5 c 1.4± ± ±0.4 a 53.1±0.7 c 1.3±0.1 1 Data are expressed as mean + SD (n=3). Values in the same column with different superscripts are significantly different(p<0.05). 22

23 . (%) Table 8. Dorsal muscle composition (% ) of the cobia fed diets containing various levels of lipids for 8 weeks 1. Dietary Lipid(%) Crude Protein Crude Lipid Moisture Ash ± ± 0.7 b 77.0±0.2 a 1.8± ± ±0.3 b 76.6±0.1 a 1.8± ± ±0.4 ab 74.7±0.3 b 1.9± ± ±0.2 a 73.6±0.3 c 1.7± ± ±0.2 a 73.5±0.4 c 1.6±0.1 1 Data are expressed as mean + SD (n=3). Values in the same column with different superscripts are significantly different(p<0.05). 23

24 . (%) Table 9. Ventral muscle composition (% ) of the cobia fed diets containing various levels of lipids for 8 weeks 1. Dietary Lipid(%) Crude protein Crude lipid Moisture Ash ± ±0.2 b 73.4±0.7 a 1.3± ± ±0.4 b 72.7±0.3 ab 1.3± ± ±0.3 b 71.7±0.4 bc 1.3± ± ±0.1 a 70.7±0.4 cd 1.4± ± ±0.2 a 69.5±0.2 d 1.4±0.9 1 Data are expressed as mean + SD (n=3). Values in the same column with different superscripts are significantly different(p<0.05). 24

25 27% 20% 25% 5% 10% 5% 10% 2.6% 20% 25% 6.0% 6.4% 5% 10 % 25% 20% 25 % 8 5% 10% aspartate transaminase (AST) 25% alanine transaminase (ALT), AST ALT ( ) 25

26 . 8 Table 10. Specific activities of aspartate transaminase (AST) and alanine transaminase (ALT) in the blood of the cobia fed diets containing various levels of lipids for 8 weeks 1. Dietary lipid (%) AST (IU/L) ALT (IU/L) ± 6.4 cd 6.8± 0.7 b ± 1.8 d 5.5± 1.0 b ± 1.6 bc 6.6 ± 1.2 b ± 9.2 ab 5.5± 1.6 b ± 7.0 a 11.9± 0.9 a 1 Data are expressed as Mean ± SD ( n=3). Values in the same column with different superscripts are significantly different(p<0.05). 26

27 / (2:1) 45% 1 15% 5% 25% 5% Jou et al.(2000) 40% (3%-18%) 3% Takeuchi et al.(1991) 10% 15% Hanley (1991) 5% Chou and Shiau (1996) 12.03% Williams (1988) 1.7% 4% 7.4% 11.2% 15% 18.8% 7.4% 11.2% 18.8% (Andrews et al.,1978) 5% n-3 27

28 1 % (Castell et al.,1994 Leu et al., 1994 Takeuchi et al.,1983 Thongrod et al., 1989 Watanabe, 1989, 1993 Yone et al.,1975) 5% 5% (FishBase, 1996) ( Shearer, 1994) (, 1987) (Oka and Ogata, 2000) (Hemre and Sandnes, 1999) (Catacutan and Coloso, 1997) (Oku and Ogata,2000) Ando and Mori (1993) (Jantrarotai et al., 1994) 25% alanine transaminase aspartate transaminase Shimeno et al.(1993) Shimeno et al. (1981) 28

29 (22%) alanine transaminase (74.7 unit/100g body weight) (Shimeno et al. 1981, 1996) 29

30 ( ) ±4.89g ±5.88g 5 8 ( 67%) Casein( 80.2%) ( ) 45 % 7.9 % 4 % 4%, 8% (Lard) (Canola) (Corn) (Soy) ( 7%) DHA(1%)(S+D) (Cod liver) 8% CMC 24% :1, 18:2n6, 18:3 20:5n3 22:6n3, ( ) 30

31 Table 11. Ingredient composition of the experimental diets (%) Composition Lard Canola Soybean Corn Soy+DHA Cod liver Fish meal Casein Alpha starch Corn starch Lard oil Canola oil Corn oil Soyben oil Soyben oil DHA Cod liver oil Vitamin mix. (a) Mineral mix. (b) Cholin chloride K 2 HPO Ca(H 2 PO 4 ) CMC (C) Metabolizable (d) Energy a Contained (as g/kg):vitamin A,600 IU/g ; Vitamin D 3,100 IU/g ; Vitamin E,50; Vitamin K 3,2.5; Thiamin,10; Riboflavin,15; Pyridoxine,7.5; Cyanocobalamin,0.025; Pantothenate,27.5; Biotin,0.5; Folic Acid, 2.5; Inositol, 100. b Contained (as g/kg):feso 4.7H 2 O,35; MgSO 4.7H 2 0,133; AlCl 3.6H 2 O,7; ZnSO 4.7H 2 0,20; CuSO 4.5H 2 O,10; NaCl,50; KI,0.05. c CMC: Carboxymethyl cellulose d Metabolizable energy : protein, Kcal/g; lipid, 8.51 Kcal/g; carbohydrate, 3.49 Kcal/g (Shiau and Huang, 1989). 31

32 . (%) Table 12. Fatty acid composition (%) in total lipid of the six oils in the experiment. Fatty acid Lard Canola Corn Soy DHA Cod liver oil C14: C16: C16: C18: C18: C18:2n C18:3n C20: C20:4n C20:5n C22: C22:5n C22:6n Saturates Monoenocs n-3 HUFA ND ND ND ND

33 . Table 13. Proximate analysis (mean ± SD n=3) of the experimental diets ( g / 100 g diet ) Dietary lipid Crude protein Lipid Moisture Ash Lard oil 44.7± ± ± ±0.1 Canola oil 44.9± ± ± ±0.2 Corn oil 44.5± ± ± ±0.1 Soybean oil 45.0± ± ± ±0.3 Soy+DHA 43.9± ± ± ±0.3 Cod liver oil 45.2± ± ± ±0.2 33

34 . (%) Table14.. Fatty acids composition (%) of the experimental diets. Fatty acid Lard Canola Corn Soybean Soy+DHA Cod liver C 14: C 16: C 16: C 18: C 18: C 18:2n C:18:3n3 ND C 20:1 ND C 20:5n C 22: C 22:6n Saturates Monoenoics n-3 HUFA n-3 HUFA/ total lipid

35 ( ) ( ) Juaneda and Rocquelin (1985) Sep-pak silica cartridges (Waters, USA) 10ml / (49:1 v/v) 7.5 ml 15 ml 2ml 0.5N KOH- methnaol 10 2ml 0.7N HCL-methnaol 3ml BF methnaol 10 3ml 2ml GC (gas chromatography) GC(HP 5890A, Japan) supelco sp ml/min 260 (PUFA-1, Supelco) ( ) SAS/PC analysis of variance ( Duncan's new multiple range test 35

36 (weight gain percent) (FCR) (PER) 1% DHA ( ) ( ) 36.8% ( ) % % % % % % % % n-3 HUFA S+D Cod liver n-3 HUFA Lard n-3 HUFA 36

37 600 Weight gain (%) Lard oil Canola oil Corn oil Soybean oil Soy+ DHA Cod liver oil Weeks. Fig 6. Percent weight gain of cobia fed diets supplemented with various lipids. 37

38 Table15. Weight gain, food conversion rate and protein efficiency ratio of the cobia fed diets supplemented with various lipids 1. Dietary Initial weight Final weight Weight gain FCR PER lipid ( g ) ( g ) ( % ) Lard ± ± 3.50 ab ± 5.18 a 1.33 ± 0.01 ab 1.67 ± 0.02 b Canola ± ± 4.70 b ± 6.22 b 1.38 ± 0.02 a 1.62 ± 0.03 b Corn ± ± 4.90 ab ± 9.41 a 1.30 ± 0.02 b 1.71 ± 0.01 a Soybean ± ± 2.20 a ± 1.40 a 1.29 ± 0.01 b 1.72 ± 0.04 a Soy+DHA ± ± 7.90 a ± 8.10 a 1.30 ± 0.03 b 1.71 ± 0.04 a Cod liver ± ± 2.60 a ± 4.40 a 1.32 ± 0.01 ab 1.68 ± 0.01 ab 1. Means (± S.E.) with different superscript letters are significantly different (p<0.05). 38

39 . 8 Table 16.. Hepatosomatic index (mean +SD n=3) of the cobia fed diets supplemented with various lipids for 8 weeks 1 Diet Hepatosomatic index (%) Lard oil 3.98± 0.13 Canola oil 4.01± 0.21 Corn oil 4.48± 0.19 Soybean oil 4.23± 0.07 Soybean oil + 1 %DHA 4.19± 0.12 Cod liver oil 4.39±

40 . (%) Table 23. Liver proximate composition (% ) of the cobia fed diets supplemented with various lipids for 8 weeks 1. Dietary lipid Crude protein Crude lipid Moisture Ash Lard oil 10.2± ±0.7 a 51.4±0.7 b 1.5±0.1 Canola oil 9.7± ±0.1 b 56.7±0.6 a 1.5±0.1 Corn oil 10.1± ±0.2 b 56.6±0.4 a 1.4±0.1 Soybean oil 9.4± ±0.4 b 55.2±0.8 a 1.7±0.2 Soy+ 1 %DHA 9.6± ±0.6 b 56.3±0.4 a 1.4±0.1 Cod liver oil 9.7± ±0.8 b 55.9±0.9 a 1.6±0.2 1 Data are expressed as Mean+SD (n=3). Values in the same column with different superscripts are significantly different(p<0.05). 40

41 . (%) Table 18. Dorsal muscle proximate composition (% ) of the cobia fed diets supplemented with various lipids for 8 weeks 1. Dietary Lipid Crude Protein Crude lipid Moisture Ash Lard oil 11.8± ± ± ±0.1 Canola oil 12.7± ± ± ±0.1 Corn oil 11.8± ± ± ±0.1 Soybean oil 12.7± ± ± ±0.2 Soy+ 1 %DHA 11.3± ± ± ±0.1 Cod liver oil 12.4± ± ± ±0.1 1 Data are expressed as Mean + SD (n=3). Values in the same column with different superscripts are significantly different(p<0.05). 41

42 . (%) Table 19. Ventral muscle proximate composition (% ) of the cobia fed diets supplemented with various lipids for 8 weeks 1. Dietary lipid Crude protein Crude lipid Moisture Ash Lard oil 12.2± ± ± ±0.1 Canola oil 12.2± ± ± ±0.2 Corn oil 12.4± ± ± ±0.1 Soybean oil 12.6± ± ± ±0.1 Soy+ 1 %DHA 11.6± ± ± ±0.1 Cod liver oil 11.2± ± ± ±0.1 1 Data are expressed as Mean + SD(n=3). Values in the same column with different superscripts are significantly different(p<0.05). 42

43 . 8 (%) Table 20. Fatty acids composition (%) of neutral lipid in the liver of the cobia fed diets supplemented with various lipids for 8 weeks. Fatty acid Lard Canola Corn Soybean Soy+DHA Cod liver C 14: C 16: C 16: C 18: C 18: C 18:2n C:18:3n C 20:1 ND ND C 20:5n C 22: C 22:6n Saturates Monoenoics n-3 HUFA

44 . 8 (%) Table 21. Fatty acids composition (%) of polar lipid in the liver of the cobia fed diets supplemented with various lipids for 8 weeks. Fatty acid Lard Canola Corn Soybean Soy+DHA Cod liver C 14: C 16: C 16: C 18: C 18: C 18:2n C:18:3n C 20:1 ND ND C 20:5n C 22: C 22:6n Saturates Monoenoics n-3 HUFA

45 . 8 (%) Table 22. Fatty acids composition (%) of neutral lipid in the ventral muscle of the cobia fed diets supplemented with various lipids for 8 weeks. Fatty acid Lard Canola Corn Soybean Soy+DHA Cod liver C 14: C 16: C 16: C 18: C 18: C 18:2n C:18:3n C 20:1 ND ND ND ND ND ND C 20:5n C 22: C 22:6n Saturates Monoenoics n-3 HUFA

46 . 8 (%) Table 23. Fatty acids composition (%) of polar lipid in the ventral muscle of the cobia fed diets supplemented with various lipids for 8 weeks. Fatty acid Lard Canola Corn Soybean Soy+DHA Cod liver C 14: C 16: C 16: C 18: C 18: C 18:2n C:18:3n C 20:1 ND ND ND ND ND 0.5 C 20:5n C 22: C 22:6n Saturates Monoenoics n-3 HUFA

47 . 8 (%) Table 24. Fatty acids composition (%) of neutral lipid in the dorsal muscle of the cobia fed diets supplemented with various lipids for 8 weeks. Fatty acid Lard Canola Corn Soybean Soy+DHA Cod liver C 14: C 16: C 16: C 18: C 18: C 18:2n C:18:3n C 20:1 ND ND ND ND ND ND C 20:5n C 22:5 ND ND ND ND ND ND C 22:6n Saturates Monoenoics n-3 HUFA

48 . 8 (%) Table 25. Fatty acids composition (%) of polar lipid in the dorsal muscle of the cobia fed diets supplemented with various lipids for 8 weeks. Fatty acid Lard Canola Corn Soybean Soy+DHA Cod liver C 14: C 16: C 16: C 18: C 18: C 18:2n C:18:3n C 20:1 ND ND ND ND ND ND C 20:5n C 22:5 ND ND ND ND ND ND C 22:6n Saturates Monoenoics n-3 HUFA

49 ( ) 1% DHA aspartate transaminase 1% DHA alanine transaminase 49

50 . 8 Table 26. Specific activities of aspartate transaminase(ast) and alanine transaminase(alt) in the blood of the cobia fed diets supplemented with various lipids for 8 weeks 1. Dietary lipid AST (IU/L) ALT (IU/L) Lard oil ±23.10 ab ±3.10 a Canola oil ±41.87 a ±1.70 b Corn oil ±24.84 bc ±2.30 bc Soybean oil ±9.20 c ±1.20 cd Soybean oil + 1% DHA 35.67±8.30 d 6.03±1.80 e Cod liver oil ±1.80 d 7.74 ±2.60 d 1. Data are expressed as Mean + SD (n=3). Values in the same column with different superscripts are significantly different (p<0.05). 50

51 14.6 % 16% 8% 8% 8 % n-3 HUFA 1.8% 1.9% 1.9% 1.9% 1%DHA 2.8% 3.8% (Owen et al., 1975 Yamada et al., 1980) n-3 HUFA 1% (Castell et al.,1994 Leu et al., 1994 Takeuchi et al., 1983 Thongrod et al., 1989 Watanabe, 1989, 1991, 1993 Yone et al.,1975) n-3hufa Greene and Selivonchick (1990) rapeseed rapeseed glucosinolates glucosinolates 51

52 (Watanabe, et al 1989 ; Tuncer and Reginal, 1992 ; Bell et al., 1994 ) aspartate transaminase n-3hufa aspartate transaminase 1% DHA aspartate transaminase DHA Ritskes-Hoitinga et al.(1998) n-3 HUFA aspartate transaminase alanine transaminase 20-30IU/L aspartate transaminase alanine transaminase alanine transaminase aspartate transaminase (Tietz, 1983) aspartate transaminase aspartate transaminase 52

53 1. 5% 20% 14.6% aspartate transaminase (AST) alanine transaminase (ALT) 4. 1% DHA 53

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