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5 7 Vol. 5 No. 7 2014 7 Journal of Food Safety and Quality Jul., 2014 谢霖霖 1, 耿越 1*, 杨维仁 2, 胥保华 (1., 250014; 2., 271018) 2 摘要 : 目的 方法 210 540, 4, 0%(A ) 3%(B ) 5%(C ) 8%(D ), 1, 50 d, 70 d 结果 (0~50 d) ω-3pufa α-ala DHA, D (P 0.01)ω-6/ω-3PUFA, D 5d 30d (P 0.01) (50~70d) ω-3 PUFA α-ala DHA ω-6/ω-3pufa (P 0.05); ω-3pufa, C (P 0.01), D (P 0.05); ω-3pufa 1.96% 1.89% 3.11% 3.28%, D (P 0.01), ω-3pufa 1.61% 1.84% 1.86% 2.27%, D (P 0.05) 结论 ω-3pufa, 3% 5% 8% 8% 关键词 : ; ; ω-3 ; Effects of different levels of rape pollen in dietary on fatty acid composition in egg yolk and tissues of laying hens XIE Lin-Lin 1, GENG Yue 1*, YANG Wei-Ren 2, XU Bao-Hua 2 (1. Provincial Key Lab of Animal Resistant Biology, College of Life Science, Shandong Normal University, Jinan 250014, China; 2. College of Animal Science and Technology, Shandong Agriculture University, Taian 281018, China) ABSTRACT: Objective To investigate the effects of dietary different levels of rape pollens on the fatty acid composition in egg yolk and tissue of laying hens. Methods 210 days old, five hundred and forty healthy Hy-Line variety brown hens which were in egg-laying peak were randomly allotted to four treatments, supplying with 0%(group A), 3%(group B), 5%(group C) and 8%(group D) rape pollens in basal diet respectively. The experiment lasted for 70 d included two phases, the first phase fed with basal diets and rape pollens (0~50 d) and the second phase fed with basal diets only (51~70 d) respectively. Results In the first stage (0~50 d), the 基金项目 : ()(200903006) (CARS-45) Fund: Supported by Special Fund for Agro-scientific Research in the Public Interest (200903006) and Earmarked Fund for Modern Agro-industry Technology Research System (CARS-45) * 通讯作者 :,, E-mail: gengy@sdnu.edu.cn *Corresponding author: GENG Yue, Professor, College of Life Science, Provincial Key Lab of Animal Resistant Biology, Shandong Normal University, Jinan 250014, China. E-mail: gengy@sdnu.edu.cn

2040 5 contents of ω-3pufa, ALA and DHA in egg yolk increased with the increasing of dietary rape pollens. But the ratio of ω-6/ω-3pufa was exact opposite and group D had significant difference from 5 d to 30 d (P<0.01). The content of ω-3pufa, ALA and DHA in egg yolk and the ratio of ω-6/ω-3pufa returned to its original level in the second stage(51~70 d). Dietary different levels of rape pollens had no effect on the fatty acid composition in brain(p 0.05). The contents of ω-3pufa in breast meat decreased with the increasing of dietary rape pollens. The contents of ω-3pufa in chicken liver were 1.96%, 1.89%, 3.11% and 3.28% respectively. Group D increased significantly(p 0.01). The contents of ω-3pufa in adipose tissue were 1.61%, 1.84%, 1.86% and 2.27% respectively. Group D increased significantly (P 0.05). Conclusion Dietary rape pollens can increase the content of ω-3pufa in egg yolk, liver and adipose tissue. The diet containing 8% of rape pollens showed the best effect among three dosages. KEY WORDS: rape pollens; egg yolk; ω-3 polyunsaturated fatty acid; diet (saturated fatty acid, SFA)(polyunsaturated fatty acid, PUFA) ω-6/ω-3pufa, [1] WHO FAO ω-6/ω-3pufa (5~10):1 [2] [3] [4], ω-3pufa ω-6/ω-3pufa, ω-3 -α- (α-linolenic acid, α-ala), 48.51% [5], SFA PUFA, V E,, ω-3pufa, 1 材料与方法 1.1 实验材料与试剂 1.1.1 实验材料 :, 95% 1.1.2 仪器与试剂 GC-2014 (); (47885-U, Sigma); ( ); ( ) 1.2 实验设计 210 540, 4, 5 27, 3, 9, 3 (A ), 3 (B ) 5 (C ) 8 (D ) 1, 50 d, 51 d, 70 d 1 1.3 饲养管理,,, 16 h (5:00 21:00),,, 1.4 样品采集 0 5 10 15 20 30 40 50 60 70 d 2, 10, 4, 24 h 41 d, 5,, -20 1.5 指标测定 1.5.1 油菜花粉和饲料中脂肪酸提取和甲酯化 0.5 g, 5 ml, 2 mol/l 0.25 ml,,, -20 [6] 1.5.2 蛋黄中脂肪酸的提取和甲酯化 1.0 g, 2 ml, 3 mol/l

7, : 2041 6 ml, 90 1 h 3 ml, 0.88% NaCl 8 ml, 4 5000 r/min 10 min, 20 表 1 实验饲粮组成及营养水平 (%, 风干基础 ) Table 1 Ingredients and composition of diets (%, air dry basis) 62.00 62.00 60.20 57.50 19.00 18.00 17.50 16.70 2.00 - - - - 3.00 5.00 8.00 3.00 3.00 3.30 3.80 9.00 9.00 9.00 9.00 1 5.00 5.00 5.00 5.00 100.00 100.00 100.00 100.00 2 (MJ/kg) 11.88 11.89 11.86 11.84 15.70 15.60 15.70 15.80 Ca 3.59 3.59 3.58 3.57 0.52 0.49 0.48 0.47 NaCl 0.38 0.38 0.37 0.37 0.80 0.76 0.74 0.71 0.35 0.35 0.34 0.33 : 1: : V A 245 000 IU, V K3 91 mg, V B1 45.5 mg, V D3 85 000 IU, V E 600 mg, V B2 175 mg, 620 mg, Cu 80 mg, Mn 1600 mg, Ca 3%~5%, NaCl 5%~9%, 6000 FTU, 10%, 160 mg Fe 740 mg Zn 800 mg P 1.5%, 28%, 2.6%,, BHT 2:, 1.5.3 蛋鸡不同组织预处理 1.5.1, 1.5.2 1.5.4 气相色谱测定条件 : KB-1701 50 m 0.32 mm 0.25 μm; : 50, 3 min, 20 /min 110, 4 /min 240, 17 min; : (N 2 ), 3 ml/min, : 50: 1; : FID ; : 250 ; : 280 ; : 1 μl 1.5.5 统计分析,, x ± SD SPSS 18.0 One-way ANOVA, LSD 2 实验结果 2.1 油菜花粉和饲料中脂肪酸分析 9.08%, α-ala 54.19%α-ALA 2.43% 2.98% 4.18% 5.55%, EPA(eicosapentaenoic acid, C20:5) DHA(docosahexaenoic acid, C22:6), ω-6/ω-3pufa 15.78 12.36 10.36 6.71 2.2 蛋黄脂肪酸气相色谱分析 1 40 d D, (C16:0), (C18:0), (C18:2, n=6)(c18:0) 2.3 饲粮中添加油菜花粉对蛋黄中脂肪酸组成的影响 2.3.1 蛋黄中 α-ala 随时间的变化趋势 (0~50 d), α-ala, 10 d α-ala, B C D 51.61% 61.29% 103.23%, α-ala (50~70 d), α-ala, C D 60 d, α-ala, 2 2.3.2 蛋黄中 DHA 随时间的变化趋势 (0~50 d), DHA, D (P 0.01), B C, DHA, 15~20 d, B C D DHA 20 d 40 d 30 d 19.31% 24.30% 34.89% 50 d DHA, 0 d, DHA α-ala, 3 2.3.3 蛋黄中 ω-3pufa 随时间的变化趋势 (0~50 d), ω-3pufa

2042 5 / Fig. 1 1 40 d D Chromatogram of fatty acids of the egg yolk of the group D on the fortieth day 1.C14:0 2.C14:1 3.C16:1 4.C16:0 5.C17:0 6.C18:1 7.C18:2 8.C18:0 9.C18:3 10.C20:4 11.C20:1 12.C20:2 13.C20:0 14.C22:6 15.C22:1 16:C22:0 表 2 蛋黄中 α-ala 含量随时间的变化 (%*, n=10) Table 2 Change on the content of α-ala in egg yolk (%*, n=10) 0 0.31±0.07 0.31±0.07 0.31±0.07 0.31±0.07 5 0.47±0.18 0.44±0.07 0.43±0.08 0.48±0.09 10 0.40±0.04 Aa 0.47±0.04 ABab 0.50±0.09 Bb 0.63±0.08 Cc 15 0.38±0.05 Aa 0.42±0.04 ABab 0.47±0.07 Bb 0.56±0.07 Cc 20 0.34±0.03 Aa 0.42±0.05 Bb 0.44±0.07 BCbc 0.55±0.07 Dd 30 0.38±0.05 Aa 0.42±0.06 ABa 0.47±0.07 Bab 0.50±0.09 Bb 40 0.44±0.03 0.46±0.09 0.46±0.06 0.47±0.07 50 0.36±0.05 Aa 0.43±0.05 ABb 0.45±0.05 Bbc 0.51±0.11 Bc 60 0.45±0.06 ABab 0.42±0.03 Aa 0.51±0.10 ABbc 0.52±0.10 Bc 70 0.41±0.07 0.40±0.03 0.39±0.07 0.40±0.05 *, : (P 0.01) (P 0.05), (P 0.05) / 表 3 蛋黄中 DHA 含量随时间的变化 (%, n=10) Table 3 Changes on the content of DHA in egg yolk (%, n=10) 0 3.21±0.33 3.21±0.33 3.21±0.33 3.21±0.33 5 3.31±0.42 Aa 3.61±0.60 ABab 3.64±0.54 ABab 4.01±0.38 Bb 10 3.46±0.33 Aa 3.75±0.25 ABa 3.79±0.47 ABa 4.19±0.44 Bb 15 3.62±0.26 Aa 3.79±0.48 ABab 3.76±0.24 ABa 4.09±0.33 Bb 20 3.64±0.33 Aa 3.83±0.43 Aa 3.81±0.26 Aa 4.31±0.35 Bb 30 3.66±0.22 Aa 3.73±0.21 Aa 3.83±0.23 Aa 4.33±0.31 Bb 40 3.31±0.20 Aa 3.62±0.33 Ab 3.99±0.30 Bb 4.15±0.36 Bb 50 3.10±0.28 Aa 3.18±0.44 ACa 3.63±0.44 BCc 4.01±0.38 Bb 60 3.10±0.23 Aa 3.28±0.45 Aab 3.47±0.30 ABb 3.71±0.34 Bb 70 3.02±0.18 3.11±0.24 3.08±0.35 3.00±0.17

7, : 2043, D (P 0.01) ω-3pufa, 20 d, B C D 20.74% 21.02% 38.07% 50 d ω-3pufa 0 d, 4 2 2.3.4 蛋黄中 ω-6/ω-3pufa 比值随时间的变化趋势 (0~50 d), ω-6/ω-3pufa, D (P 0.01)ω-6/ω-3PUFA, 40 d, C, 6.36 4.82 40~50 d, ω-6/ω-3pufa, 50 d, ω-6/ω-3pufa, 0 d, 5 3 2.4 蛋鸡不同组织脂肪酸组成测定 5.75% 7.52% 4.41% 4.83%, C (P 0.01) PUFA, 8% DHA, 74.84%, ω-6/ω-3pufa 表 4 蛋黄中 ω-3pufa 含量随时间的变化 (%, n=10) Table 4 Changes of the content of ω-3pufa in egg yolk (%, n=10) / 0 3.52±0.33 3.52±0.33 3.52±0.33 3.52±0.33 5 3.78±0.43 Aa 4.05±0.59 ABab 4.07±0.51 ABab 4.49±0.42 Bb 10 3.86±0.34 Aa 4.21±0.26 Aab 4.29±0.48 Ab 4.82±0.45 Bb 15 4.00±0.28 Aa 4.22±0.49 Aa 4.23±0.26 ABa 4.65±0.31 Bb 20 3.98±0.34 Aa 4.25±0.42 Aa 4.26±0.28 Aa 4.86±0.39 Bb 30 4.05±0.23 Aa 4.15±0.24 Aab 4.30±0.25 Ab 4.83±0.34 Bb 40 3.75±0.19 Aa 4.08±0.28 Ab 4.45±0.27 Bb 4.62±0.36 Bb 50 3.46±0.31 Aa 3.61±0.42 Aa 4.08±0.46 Ba 4.52±0.30 Bb 60 3.55±0.23 Aa 3.70±0.44 Aab 3.98±0.33 ABb 4.23±0.38 Bb 70 3.43±0.14 3.51±0.24 3.47±0.35 3.40±0.14 2 ω-3pufa Fig. 2 The increasing rate of ω-3pufa in egg yolk among rape pollens dietary groups : ω-3pufa (%)=( ω-3pufa ω-3pufa )/ ω-3pufa 100%

2044 5 表 5 蛋黄中 ω-6/ ω-3pufa 比值随时间的变化 (n=10) Table 5 Change on the ratio of ω-6/ ω-3pufa in egg yolk (n=10) / 0 6.36±0.67 6.36±0.67 6.36±0.67 6.36±0.67 5 6.36±0.60 Aa 5.88±0.81 ABab 5.64±0.73 ABb 5.36±0.51 Bb 10 6.00±0.54 Aa 5.51±0.21 ABb 5.42±0.62 ABbc 5.02±0.44 Bc 15 5.94±0.43 Aa 5.49±0.57 ABb 5.50±0.36 ABb 5.00±0.36 Bc 20 5.73±0.50 Aa 5.54±0.61 ABa 5.51±0.49 ABa 4.93±0.42 Bb 30 5.75±0.30 Aa 5.54±0.28 Aab 5.42±0.34 Ab 4.86±0.34 Bb 40 6.10±0.46 Aa 5.28±0.73 Bb 4.82±0.58 Bb 4.91±0.53 Bb 50 6.70±0.78 Aa 6.40±0.65 ACa 5.68±0.68 BCc 5.01±0.54 Bb 60 6.80±0.49 Aa 6.28±0.78 ABb 5.82±0.41 Bab 5.62±0.46 Ba 70 6.82±0.45 6.53±0.30 6.74±0.78 6.83±0.43 3 ω-6/ ω-3pufa Fig. 3 Changes on the ratios of ω-6/ω-3pufa in egg yolk, B (arachidonic acid, AA), C D 42.7% 51.2%, (P 0.05); (P 0.05);, ω-3pufa, ω-6pufa ω-6/ω-3pufa, C D (P 0.01); DHA, (P 0.05) D ω-3pufa 67.35% 40.99%, ω-6/ω-3pufa 7.50 10.76 6 3 讨论 3.1 饲粮中添加不同水平油菜花粉对蛋黄脂肪酸组成的影响 ω-3pufa α-ala EPA DHA EPA, [7] α-ala, EPA DHA, EPA (0~50 d), α-ala DHA, α-ala

7, : 2045 Table 6 表 6 蛋鸡组织中脂肪酸组成的变化 (%, n=5) Change on the fatty acid composition in tissue of laying hens (%, n=5) C18:3 α-ala 0.40±0.10 a 0.69 ±0.27 b 0.53±0.06 ab 0.57±0.13 ab C20:4 AA 3.89±0.82ABa 2.98 ±1.45Aa 5.55±1.04Bb 5.88±1.39Bb C22:6 DHA 1.55±0.34 Aa 1.21 ±0.56 Aa 2.58±0.55 Bb 2.71±0.26 Bb ω-3pufa 1.96±0.27 Aa 1.89 ±0.80 Aa 3.11±0.56 Bb 3.28±0.35 Bb ω-6pufa 19.44±0.61 ac 19.07 ±5.77 a 24.24±2.53 bc 24.58±3.44 b ω-6/ω-3pufa 10.11±1.61 ACa 10.48 ±1.39 Aa 7.91±0.92 BCb 7.50±0.80 Bb C18:3 α-ala 0.31±0.10 0.28±0.04 0.31±0.10 0.41±0.24 C20:4 AA 9.69±0.59 9.87±0.66 9.87±1.25 9.03±0.94 C22:6 DHA 9.36±0.63 10.11±0.95 9.41±1.12 9.32±1.23 ω-3pufa 9.67±0.59 10.39±0.96 9.71±1.09 9.65±1.15 ω-6pufa 22.49±0.62 21.58±0.52 22.12±0.70 21.71±1.42 ω-6/ω-3pufa 2.33±0.14 2.10±0.25 2.30±0.31 2.29±0.38 C18:3 α-ala 0.75±0.14 0.68±0.05 NF NF C20:4 AA 9.77±0.99 8.73±0.19 8.9±1.08 10.46±2.24 C22:6 DHA 3.14±0.25 Aa 3.03±0.22 Aa 2.27±0.26 Bb 2.74±0.49 Aba ω-3pufa 3.89±0.39 Aa 3.71±0.27 Aa 2.27±0.26 Bb 2.74±0.49 Bb ω-6pufa 26.93±0.82 Aa 26.40±0.56 Aa 37.20±0.90 Bb 37.07±2.31 Bb ω-6/ω-3pufa 6.97±0.54 Aa 7.14±0.54 Aa 16.59±1.96 Ba 13.88±2.68 Bb C18:3 α-ala 1.24±0.21 Aa 1.46±0.37 ABab 1.34±0.13 ABa 1.77±0.24 Bb C20:4 AA 0.76±0.36 0.77±0.23 1.00±0.24 1.00±0.15 C22:6 DHA 0.37±0.19 0.38±0.10 0.52±0.14 0.50±0.10 ω-3pufa 1.61±0.17 Aa 1.84±0.37 ABa 1.86±0.19 ABa 2.27±0.30 Bb ω-6pufa 20.84±1.68 Aa 22.71±1.98 ABab 22.41±1.07 ABab 24.13±0.81 Bb ω-6/ω-3pufa 12.96±0.81 a 12.61±2.06 ab 12.15±0.95 ab 10.76±1.51 b : NF=no found 10 d, DHA 20 d α-ala, α-ala 0.12% 3.50%, α-ala, 0.50% 5.69%, 14 d DHA,, DHA [8] 10 d [9] ω-3pufa α-ala 2.43% 5.55%, 10 d D α-ala, 0.63% α-ala,, α-ala ω-6/ω-3pufa, D (P 0.01) WHO ω-6/ω-3pufa (5~10):1; ω-6/ω-3pufa 9.8:1 2.3:1 [10] ;

2046 5 ω-6/ω-3pufa (4~6):1 [11] C D ω-6/ω-3pufa (4~6):1, ω-6/ω-3pufa, 60 d, C D α-ala, DHA ω-3pufa 50 d (P 0.05), ω-6/ω-3pufa 50 d (P 0.01) 70 d, C D α-ala DHA ω-3pufa ω-6/ω-3pufa (P 0.05) α-ala DHA ω-3pufa ω-6/ω-3pufa, 2, 3.2 饲粮中添加不同水平油菜花粉对蛋鸡组织中脂肪酸组成的影响 α-ala DHA ω-3pufa, D 42.50% 74.84% 67.35%, [12], (AA)(P 0.05), α-ala, α-ala, AA (P 0.05) [8],,,, (P 0.05), [13], α-ala, α-ala α-ala (P 0.05), DHA, 10% DHA, [13], α-ala DHA, ω-6/ω-3pufa C 16.59%±1.96%, ω-3pufa ω-3pufa (P 0.05), ω-6/ω-3pufa (P 0.05) [14] α-ala, α-ala,, [13],,,, [15], [16], ω-3pufa ω-3pufa,,, ω-3pufa,, 4 结论, ω-3pufa α-ala DHA, ω-6/ω-3, 3% 5% 8% 8%, ω-3pufa ω-3pufa, ω-3pufa 参考文献 [1] Sioen I, Hacquebard M, Hick G, et al. Effect of ALA-enriched food supply on cardiovascular risk factors in males [J]. Lipids, 2009, 44(7): 603 611. [2] Sugano M, Hirahara F. Polyunsaturated fatty acids in the food chain in Japan [J]. Am J Clin Nutr, 2000, 71(1): 189 196. [3],,. [J]., 2002, (12): 8 9,11. Wang LH, Huo GC, Yang LJ. Deposition effect of α-linolenic acid from flaxseed in egg yolk [J]. China Feed 2002, (12): 8 9, 11. [4]. [D]. :, 2009. Ma HB. Effect of dietary oil sources on egg quality and the content of polyunsaturated fatty acids of yolk [D]. Beijing: Chinese Academy of Agricultural Sciences, 2009. [5],,,. [J]., 2005, 27(4): 351 352.

7, : 2047 Tu WL, Geng Y, Zhang JJ, et al. Analysis of fatty acid composition of eight kinds of pollens [J]. Acta Nutrimenta Sinica, 2005, 27(4): 351 352. [6] Wang Y, Sunwoo H, Cherian G, et al. Fatty acid determination in chicken egg yolk: a comparison of different methods [J]. Poultry Sci, 2000, 79(8): 1168 1171. [7],,,. ω-3 ω-6 [J]., 2003, 26(1): 65 68, 72. Zang SM, Li TZ, He WH, et al. The effects of Perilla frutescens seeds on fatty acid composition and ω-3, ω-6 of egg yolk [J]. J Agric Univ Hebei, 2003, 26(1): 65 68, 72. [8],,,. n-3 α- [J]., 2000, 5(1): 117 122. Yang CX, Ji C, Ding LM, et al. n-3 Fatty acid metabolism and effects of α-linolenic acid on enriching n-3 FA eggs [J]. J China Agric Univ, 2000, 5(1): 117 122. [9]. [J]., 2000, 13(4): 84 89. Xia ZS. Effect of feeding different oils to laying hens on the lipid content and fatty acid composition in egg yolk [J]. Southwest China J Agric Sci, 2000, 13(4): 84 89. [10] Kris-Etherton PM, Taylor DS, Yu-Poth S, et al. Polyunsaturated fatty acids in the food chain in the United States [J]. Am J Clin Nutr, 2000, 71(1): 179S 188S. [11],. [J]., 2003, 28(1): 31 34. Su YX, Guo Y. A review of dietary fatty acid composition and recommended optimal ratio [J]. China Oil Lipid, 2003, 28(1): 31 34. [12],,,. [J]., 2010, 42(6): 40 43. Chen JX, Lu J, Xia ZS, et al. Influences of high level linolenic acid on lipid content in egg yolk and lipid metabolism in laying hens [J]. Anim Husbandry Vet Med, 2010, 42(6): 40 43. [13]. n-3 [D]. :, 2000. Wang K. Enrichment of n-3 polyunsaturated fatty acid in egg yolk and tissues and its effect on lipid metabolism in laying hens [D]. Beijing: Chinese Academy of Agricultural Sciences, 2000. [14],,,. ω-6/ω-3 [J]., 2009, 21(4): 525 533. Qi KK, Zhao GP, Chen JL, et al. Effect of dietary ω-6/ω-3 on meat quality, serum lipid and fatty acid profiles of arbor acres briolers [J]. Chin J Anim Nutr, 2009, 21(4): 525 533. [15],,,. [J]., 2007, (4): 37 38. Liu WG, Yao YB, Zhang XY, et al. Influences of bee pollen on the production of laying hens [J]. Cereal Feed Ind, 2007, (4): 37 38. [16],,,. [J]., 2007, 21(6): 1 4. Wang Y, Zheng YM, Wu W, et al. Influences of bee pollen in diet on the egg quality [J]. J Anhui Sci Technol Univ, 2007, 21(6): 1 4. 作者简介 ( 责任编辑 : 杨翠娜 ) 谢霖霖, 硕士, 主要研究方向为食品营养与食品功能因子 E-mail: w3fattyacids@163.com 耿越, 教授, 主要研究方向为食品营养与功能因子 E-mail: gengy@sdnu.edu.cn