Chinese Journal of Animal Nutrition >

2025 , Vol. 37 >Issue 4: 2576 - 2586

DOI: https://doi.org/10.12418/CJAN2025.216

RESEARCH PAPER

Effects of Dietary Supplementation with Schizochytrium Meal on Growth Performance, Non-Specific Immunity and Antioxidant Capacity of Lateolabrax maculatus

  • LIU Xiaocheng , 1 ,
  • XIONG Yi 1 ,
  • ZHONG Huichang 2 ,
  • ZHANG Chunxiao 1 ,
  • WANG Ling 1 ,
  • SONG Kai 1 ,
  • MA Ruijuan 1 ,
  • LI Xueshan 1 ,
  • LU Kangle , 1, *
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  • 1 National Key Laboratory of Biological Breeding for Mariculture, College of Fisheries, Jimei University, Xiamen 361021, China
  • 2 Xiamen Huison Biotech Co., Ltd., Xiamen 361021, China
* professor, E-mail:

Received date: 2024-09-29

  Online published: 2025-04-15

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Abstract

The aim of this experiment was to investigate the effects of dietary supplementation with Schizochytrium meal on the growth performance, non-specific immunity and antioxidant capacity of Lateolabrax maculatus, and to provide a theoretical basis for the application of Schizochytrium meal in the diet of Lateolabrax japonicus. On the basis of a basal diet formula with 44% crude protein and 11% crude lipid, five experimental diets were formulated with 0 (control), 0.8%, 1.6%, 2.4% and 3.2% Schizochytrium meal to replace the same amount of wheat flour. Five experimental diets were fed to Lateolabrax japonicus with the same initial size [body weight (2.81±0.02) g] for 60 days. The results showed that the weight gain rate and specific growth rate of Lateolabrax japonicus increased at first and then decreased with the increase of Schizochytrium meal, and dietary supplementation with 1.6% and 3.2% Schizochytrium meal significantly increased the weight gain rate and specific growth rate compared with the control group (P<0.05). Compared with the control group, dietary supplementation with different levels of Schizochytrium meal significantly decreased the crude lipid content in the muscle of Lateolabrax japonicus (P<0.05). The serum triglyceride content, alanine transaminase and aspartate transaminase activities of Lateolabrax japonicus fed diets supplemented with 0.8%, 1.6% and 3.2% Schizochytrium meal were significantly lower than those of the control group (P<0.05). The serum acid phosphatase activity of Lateolabrax japonicus fed diets supplemented with 1.6%, 2.4% and 3.2% Schizochytrium meal was significantly higher than that of the control group (P<0.05), and dietary supplementation with 2.4% Schizochytrium meal also significantly increased the serum immunoglobulin M content of Lateolabrax japonicus (P<0.05). Compared with the control group, the activities of catalase, superoxide dismutase and glutathione peroxidase in the liver of Lateolabrax japonicus were significantly increased (P<0.05), while the content of malondialdehyde in the liver was significantly decreased by adding 1.6% Schizochytrium meal in the diet (P<0.05). In conclusion, dietary supplementation with 1.6% Schizochytrium meal can promote the growth of Lateolabrax japonicus, and at the same time improve the body non-specific immunity and liver antioxidant capacity of Lateolabrax japonicus.

Key words： Schizochytrium meal; Lateolabrax maculatus; growth performance; non-specific immunity; antioxidant capacity

Cite this article

LIU Xiaocheng , XIONG Yi , ZHONG Huichang , ZHANG Chunxiao , WANG Ling , SONG Kai , MA Ruijuan , LI Xueshan , LU Kangle . Effects of Dietary Supplementation with Schizochytrium Meal on Growth Performance, Non-Specific Immunity and Antioxidant Capacity of Lateolabrax maculatus[J]. Chinese Journal of Animal Nutrition, 2025 , 37(4) : 2576 -2586 . DOI: 10.12418/CJAN2025.216

裂壶藻是一种海洋微藻,其细胞内含有大量的油脂,其中二十二碳六烯酸(DHA)含量最多[1]。DHA是一种是重要的n-3多不饱和脂肪酸,能促进婴幼儿大脑和视网膜的发育及成熟,并且对降低血脂、预防心血管疾病等有明显作用,是现代营养学极为推崇的营养素[2-3]。此外,提取裂壶藻中的DHA作为食品添加剂添加到牛奶、米粉等婴幼儿食品和食用油等食品中,或作为保健食品也极为普遍[4]。由于裂壶藻生长速度快,可通过异养方式高效培养,并且细胞内的DHA含量高,使其成为微生物发酵生产DHA的优良菌种[5]。随着这项产业的迅速发展,提取DHA后产生的裂壶藻发酵副产物产量很大,据推测,我国每年有数万吨的产量。裂殖壶藻发酵培养提取DHA后的膏状物浓缩物,称之为裂壶藻粕,其含蛋白质、脂肪、碳水化合物、矿物质和维生素等营养成分,如何处理和有效利用这些裂壶藻粕已成为需要解决的问题。花鲈(Lateolabrax maculatus)又称海鲈、七星鲈,隶属于鲈形目,鮨科,花鲈属,近年来因其肉质鲜美、营养丰富、生长速度快等特点养殖产量不断上涨[6]。2022年我国花鲈的年产量达21.8万t,仅次于大黄鱼和石斑鱼,是一种重要的经济鱼类[7]。花鲈养殖主要集中在东南沿海地区,而裂壶藻发酵DHA产业也多集中于南方地区。因此,花鲈作为裂壶藻粕应用的研究对象较为适宜。本试验在测定裂壶藻粕营养组成的基础上,将其添加到花鲈饲料中,研究其对花鲈生长性能、非特异性免疫及抗氧化能力的影响,为了裂壶藻发酵副产物在水产养殖中的应用提供参考。

1 材料与方法

1.1 试验饲料

基于花鲈的营养需求特点,以鱼粉、豆粕、鸡肉粉和谷朊粉为主要蛋白质源,鱼油、豆油和卵磷脂为主要脂肪源,设计含有44%粗蛋白质和11%粗脂肪的基础饲料配方。分别以0(对照)、0.8%、1.6%、2.4%、3.2%(以干物质计)的裂壶藻粕(裂壶藻发酵生产DHA后的副产物,为膏状)等量替代基础饲料配方中的面粉,配制5种试验饲料,其组成及营养水平见表1。
表1 试验饲料组成及营养水平(干物质基础)

Table 1 Composition and nutrient levels of experimental diets (DM basis) %

项目
Items		 裂壶藻粕添加量Schizochytrium meal additive amount/%
0 0.8 1.6 2.4 3.2
原料Ingredients
鱼粉Fish meal 26.85 26.85 26.85 26.85 26.85
鸡肉粉Chicken meal 8.47 8.47 8.47 8.47 8.47
谷朊粉Gluten flour 7.30 7.30 7.30 7.30 7.30
豆粕Soybean meal 25.00 25.00 25.00 25.00 25.00
α-淀粉α-starch 3.00 3.00 3.00 3.00 3.00
面粉Wheat flour 20.03 19.23 18.43 17.63 16.83
鱼油Fish oil 2.00 2.00 2.00 2.00 2.00
豆油Soybean oil 2.00 2.00 2.00 2.00 2.00
卵磷脂Lecithin 2.00 2.00 2.00 2.00 2.00
维生素C Vitamin C 0.05 0.05 0.05 0.05 0.05
维生素预混料Vitamin premix1) 0.30 0.30 0.30 0.30 0.30
矿物质预混料Mineral premix2) 0.50 0.50 0.50 0.50 0.50
氯化胆碱Choline chloride 0.50 0.50 0.50 0.50 0.50
磷酸二氢钙Ca(H2PO4)2 2.00 2.00 2.00 2.00 2.00
裂壶藻粕Schizochytrium meal 0.80 1.60 2.40 3.20
合计Total 100.00 100.00 100.00 100.00 100.00
营养水平Nutrient levels3)
粗蛋白质Crude protein 44.93 44.91 44.85 44.86 44.82
粗脂肪Crude lipid 10.47 10.99 11.07 11.07 11.54

1)维生素预混料为每千克饲料提供Vitamin premix provided the following per kg of diets:硫胺素 thiamin 25 mg,核黄素 riboflavin 45 mg,盐酸吡哆醇 pyridoxine hydrochloride 20 mg,VB12 0.1 mg,VK3 10 mg,肌醇 inositol 800 mg,泛酸钙calcium pantothenate 60 mg,烟酰胺 nicotinamide 200 mg,叶酸 folic acid 20 mg,生物素 biotin 1.2 mg,维生素A乙酸酯 VA acetate 32 mg,VD3 5 mg,α-生育酚 α-tocopherol 120 mg,乙氧基喹啉 ethoxyquinoline 150 mg,玉米淀粉 corn starch 1 511.7 mg。

2)矿物质预混料为每千克饲料提供 Mineral premix provided the following per kg of diets:Ca 1 150 mg,K 180 mg,Mg 45 mg,Mn 9.5 mg,Cu 7.5 mg,Co 1.25 mg,I 0.16 mg,Se 0.25 mg。

3)营养水平均为实测值。Nutrient levels were measured values.

饲料原料粉碎后过60目筛,按表1准确称取各原料后逐级混合均匀,裂壶藻粕溶于鱼油后再与其他原料混合,确保均匀分布。加入适量水经搅拌机(BFJ-2018-15C,济南泰诺机械有限公司)混合均匀,用挤条机(F-76,广州华工光机电科技有限公司)挤压成条,最终经造粒机(GY-500,常州市倍成干燥设备工程有限公司)制成粒径为2 mm的颗粒饲料,45 ℃烘箱中烘干后,置于-20 ℃冰箱中储存备用。

1.2 试验鱼及饲养管理

本试验所用花鲈购于漳州某商业育苗场,在厦门市集美大学水产试验场的室内循环水养殖系统中进行养殖试验。花鲈鱼苗运至水产试验场后于养殖池中暂养14 d,暂养期间投喂商品饲料。暂养结束后,挑选300尾体质健康、规格一致的花鲈[体重(2.81±0.02) g],随机分配到15个循环水养殖桶中,每桶20尾,每组分配3个养殖桶(重复),养殖周期为60 d。试验期间每日定时(08:30、17:30)进行表观饱食投喂2次,记录摄食量并适量换水。养殖期间保持以下条件:光周期12 h/12 h,水温(27±2) ℃,溶解氧含量≥5 mg/L,氨氮含量<0.1 mg/L,pH 7.5~8.5。

1.3 样品采集

样品采集过程按照集美大学动物管理委员会的标准(No.2019-32)执行。养殖试验结束后,试验鱼禁食24 h,然后将所有试验鱼用MS-222(70 mg/L)麻醉后称重并测量体长。每桶随机取10尾试验鱼,使用1 mL无菌注射器从尾静脉抽血,血液样品在4 ℃环境静置6 h后以1 000×g离心10 min,分离血清,置于-80 ℃冰箱待用;试验鱼抽血后立即进行解剖,称量并记录鱼体内脏团、肝脏和腹部脂肪的重量,将肝脏于-80 ℃冰箱保存待测抗氧化指标,同时取3尾鱼的背部肌肉用于分析肌肉营养成分;此外,每桶再随机取2尾试验鱼,保存至-20 ℃冰箱,用于分析全鱼营养成分。

1.4 指标测定

1.4.1 生长性能和形体指标测定

生长性能和形体指标计算公式如下:
存活率(SR,%)=(末尾数/初尾数)×100;
增重率(WG,%)=[(末均重-初均重)/初均重]×100;
特定生长率(SGR,%/d)=[(ln末均重-ln初均重)/试验天数]×100;
摄食量(FI,g/尾)=每桶总摄食量/末尾数;
饲料系数(FCR)=摄食总量/(末总重-初总重);
蛋白质效率(PER)=(末总重-初总重)/蛋白质摄入量;
肝体比(HSI,%)=(肝脏重/鱼体重)×100;
脏体比(VSI,%)=(内脏团重/鱼体重)×100;
肥满度(CF,%)=(鱼体重/鱼体长3)×100;
腹脂率(IFR,%)=(腹部脂肪质量/鱼体重)×100。

1.4.2 裂壶藻粕、饲料、全鱼和肌肉营养成分分析

参考AOAC(1995)[8]的方法测定裂壶藻粕、饲料、全鱼和肌肉的水分、粗蛋白质和粗脂肪含量以及裂壶藻粕的粗灰分含量。参考宋泽等[1]的研究,采用高效液相色谱法测定裂壶藻粕中藻胶多糖含量;采用气相色谱法测定裂壶藻粕中DHA与脂肪酸含量;参照《饲料中挥发性盐基氮的测定》(GB/T 32141—2015)[9]测定裂壶藻粕中挥发性盐基氮含量。

1.4.3 血清生化指标测定

血清谷丙转氨酶(ALT,C009-2-1)、谷草转氨酶(AST,C010-2-1)活性及甘油三酯(TG,A110-1-1)、葡萄糖(GLU,A154-1-1)和总胆固醇(T-CHO,A111-1-1)含量均采用相应试剂盒(南京建成生物工程研究所)测定,具体测定步骤参照试剂盒说明书。

1.4.4 血清免疫指标测定

血清酸性磷酸酶(ACP,A060-2)、碱性磷酸酶(AST,A059-2)、溶菌酶(LZM,A050-1-1)活性均采用相应试剂盒(南京建成生物工程研究所)测定,免疫球蛋白M(IgM,H109-1-2)和补体4(C4,H186-2-2)含量采用鱼类专用酶联免疫吸附测定(ELISA)试剂盒(南京建成生物工程研究所)测定,具体测定步骤参照试剂盒说明书。

1.4.5 肝脏抗氧化指标测定

肝脏过氧化氢酶(CAT,A007-1-1)、超氧化物歧化酶(SOD,A001-3-1)、谷胱甘肽过氧化物酶(GSH-Px,A005-1)活性和丙二醛(MDA,A003-1)含量均采用相应试剂盒(南京建成生物工程研究所)测定,具体测定步骤参照试剂盒说明书。

1.5 数据统计与分析

试验数据以平均值±标准误(mean±SE)表示。采用SPSS 27.0软件进行单因素方差分析(one-way ANOVA),并采用Duncan氏法进行多重比较,P<0.05为显著性水平。

2 结果

2.1 裂壶藻粕的营养组成和脂肪酸组成

本试验所用裂壶藻粕含有59.6%的水分,其粗蛋白质含量为18.8%,粗脂肪含量为12.5%,藻胶多糖含量为11.3%,DHA和挥发性盐基氮含量分别为5.63%和6.08%(表2);裂壶藻粕的脂肪酸中有25.9%的饱和脂肪酸、1.61%的单不饱和脂肪酸、71.43%的n-3多不饱和脂肪酸和0.93%的n-6多不饱和脂肪酸(表3)。
表2 裂壶藻粕的营养组成(干物质基础)

Table 2 Nutrition composition of Schizochytrium meal (DM basis)

项目Items 含量Content
粗蛋白质Crude protein/% 18.83
粗脂肪Crude lipid/% 12.55
粗灰分Ash/% 14.51
藻胶多糖Algin polysacchride/% 11.32
二十二碳六烯酸DHA/% 5.63
挥发性盐基氮
Volatile basic nitrogen/(mg/100 g)		 6.08
表3 裂壶藻粕的脂肪酸组成(占总脂肪酸的百分比)

Table 3 Fatty acid composition of Schizochytrium meal (percentage of total fatty acids) %

脂肪酸Fatty acids 含量Content
ΣSFA 25.90
ΣMUFA 1.61
Σn-3PUFA 71.43
Σn-6PUFA 0.93
Σn-3PUFA/Σn-6PUFA 76.81

ΣSFA:总饱和脂肪酸 total saturated fatty acid; ΣMUFA:总单不饱和脂肪酸 total monounsaturated fatty acid; Σn-3PUFA:总n-3多不饱和脂肪酸 total n-3 polyunsaturated fatty acids; Σn-6PUFA:总n-6多不饱和脂肪酸 total n-6 polyunsaturated fatty acids。

2.2 裂壶藻粕对花鲈生长性能和形体指标的影响

由表4可知,花鲈的增重率和特定生长率随着裂壶藻粕添加量的提高呈现先升高后降低的趋势,均在添加量为1.6%时达到最高,且与对照组相比,饲料中添加1.6%和3.2%的裂壶藻粕显著提高了花鲈的增重率和特定生长率(P<0.05);与对照组相比,饲料中添加不同水平裂壶藻粕对花鲈的脏体比无显著影响(P>0.05);与对照组相比,饲料中添加1.6%和2.4%的裂壶藻粕显著提高了花鲈的腹脂率(P<0.05);花鲈的存活率、摄食量、饲料系数、蛋白质效率、肝体比和肥满度各组间均无显著差异(P>0.05)。
表4 裂壶藻粕对花鲈生长性能和形体指标的影响

Table 4 Effects of Schizochytrium meal on growth performance and physical indices of Lateolabrax maculatus

项目
Items		 裂壶藻粕添加量Schizochytrium meal additive amount/%
0(对照Control) 0.8 1.6 2.4 3.2
初始均重IBW/g 2.81±0.02 2.81±0.02 2.81±0.02 2.81±0.02 2.81±0.02
终末均重FBW/g 52.07±3.47a 56.19±2.49ab 63.66±1.43c 54.82±1.88ab 56.88±1.59b
存活率SR/% 93.33±1.67 98.33±1.67 96.67±1.67 95.00±2.89 96.67±1.67
增重率WGR/% 1 753.12±71.27a 1 899.54±51.21ab 2 165.42±50.92c 1 851.03±38.70ab 1 924.13±32.61b
特定生长率SGR/(%/d) 5.21±0.07a 5.34±0.05ab 5.57±0.02c 5.30±0.04ab 5.37±0.03b
摄食量FI/(g/尾) 60.98±4.57 60.85±0.99 63.14±3.17 60.76±1.77 57.92±1.22
饲料系数FCR 1.11±0.01 1.08±0.03 1.07±0.04 1.07±0.03 1.04±0.02
蛋白质效率PER 2.00±0.02 2.06±0.05 2.08±0.09 2.10±0.06 2.14±0.08
肝体比HSI/% 1.16±0.05 1.15±0.06 1.10±0.03 1.07±0.01 1.05±0.02
脏体比VSI/% 10.54±0.15ab 11.16±0.25b 11.13±0.07b 11.17±0.14b 10.44±0.26a
腹脂率IFR/% 5.07±0.07a 5.47±0.10ab 5.85±0.08b 5.85±0.17b 5.44±0.36ab
肥满度CF/% 1.68±0.03 1.68±0.01 1.68±0.03 1.75±0.03 1.71±0.02

同行数据肩标不同小写字母表示差异显著(P<0.05)。下表同。

Values in the same row with different small letter superscripts mean significant difference (P<0.05). The same as below.

2.3 裂壶藻粕对花鲈全鱼和肌肉营养组成的影响

由表5可知,花鲈全鱼与肌肉的水分和粗蛋白质含量各组间无显著差异(P>0.05);饲料中裂壶藻粕添加量为2.4%时,花鲈全鱼粗脂肪含量显著高于对照组和其他添加裂壶藻粕组(P<0.05);此外,与对照组相比,饲料中添加不同水平裂壶藻粕均显著降低了花鲈肌肉的粗脂肪含量(P<0.05)。
表5 裂壶藻粕对花鲈全鱼和肌肉营养组成的影响

Table 5 Effects of Schizochytrium meal on nutrition composition in whole body and muscle of Lateolabrax maculatus

项目
Items		 裂壶藻粕添加量Schizochytrium meal additive amount/%
0(对照Control) 0.8 1.6 2.4 3.2
全鱼Whole body
水分Moisture 69.91±0.25 70.01±0.49 70.20±0.46 69.30±0.28 69.92±0.28
粗蛋白质Crude protein 17.24±0.19 17.42±0.49 17.31±0.19 17.45±0.27 17.37±0.44
粗脂肪Crude lipid 8.99±0.21a 8.70±0.07a 9.00±0.23a 9.78±0.16b 8.87±0.14a
肌肉Muscle
水分Moisture 77.39±0.40 77.24±0.08 77.54±0.22 77.69±0.08 77.47±0.13
粗蛋白质Crude protein 21.02±0.50 21.48±0.16 21.29±0.26 21.05±0.10 21.29±0.06
粗脂肪Crude lipid 0.99±0.03c 0.83±0.04b 0.67±0.02a 0.68±0.02a 0.65±0.02a

2.4 裂壶藻粕对花鲈血清生化指标的影响

由表6可知,与对照组相比,饲料中添加不同水平裂壶藻粕均显著降低了花鲈血清中甘油三酯含量(P<0.05);与对照组相比,0.8%、2.4%、3.2%裂壶藻粕添加组花鲈血清中葡萄糖含量显著降低(P<0.05);与对照组相比,各裂壶藻粕添加组花鲈血清中谷丙转氨酶活性均显著降低(P<0.05),同时0.8%、1.6%、3.2%裂壶藻粕添加组花鲈血清谷草转氨酶活性也显著降低(P<0.05);此外,花鲈血清中总胆固醇含量各组间均无显著差异(P>0.05)。
表6 裂壶藻粕对花鲈血清生化指标的影响

Table 6 Effects of Schizochytrium meal on serum biochemical indices of Lateolabrax maculatus

项目
Items		 裂壶藻粕添加量Schizochytrium meal additive amount/%
0(对照Control) 0.8 1.6 2.4 3.2
甘油三酯TG/(mmol/L) 7.29±0.11a 5.63±0.06b 4.55±0.11bc 3.61±0.78c 3.23±0.52c
总胆固醇T-CHO/(mmol/L) 6.36±0.57 6.05±0.88 5.73±0.43 4.67±0.60 6.65±1.30
葡萄糖GLU/(mmol/L) 8.04±0.40a 6.21±0.31b 8.77±0.56a 5.54±0.31b 5.23±0.13b
谷丙转氨酶ALT/(U/L) 27.19±1.14a 20.81±1.54b 15.18±1.77c 9.27±0.42d 10.48±0.94d
谷草转氨酶AST/(U/L) 13.88±2.18a 9.25±0.38b 10.37±0.57b 15.07±0.48a 10.28±0.70b

2.5 裂壶藻粕对花鲈血清免疫指标的影响

由表7可知,与对照组相比,1.6%、2.4%、3.2%裂壶藻粕添加组花鲈血清中酸性磷酸酶活性显著升高(P<0.05),且2.4%裂壶藻粕添加组花鲈血清中免疫球蛋白M含量亦显著升高(P<0.05);饲料中添加不同水平裂壶藻粕对花鲈血清中碱性磷酸酶、溶菌酶活性和补体4含量均无显著影响(P>0.05)。
表7 裂壶藻粕对花鲈血清免疫指标的影响

Table 7 Effects of Schizochytrium meal on serum immune indices of Lateolabrax maculatus

项目
Items		 裂壶藻粕添加量Schizochytrium meal additive amount/%
0(对照Control) 0.8 1.6 2.4 3.2
酸性磷酸酶ACP/(U/dL) 6.55±0.22a 8.29±0.35ab 9.80±0.69b 9.08±1.17b 9.41±0.19b
碱性磷酸酶AKP/(U/dL) 4.13±0.42 4.12±0.18 5.55±0.40 5.26±0.84 5.62±0.82
免疫球蛋白M IgM/(μg/mL) 322.66±11.95a 354.13±27.71a 388.87±23.23a 570.65±63.48b 448.07±69.95ab
补体4 C4/(μg/mL) 129.74±26.97 160.37±32.00 183.62±3.25 171.04±8.14 184.35±9.24
溶菌酶LZM/(μg/mL) 19.66±1.24 20.02±1.83 18.04±0.99 18.85±0.97 18.70±1.62

2.6 裂壶藻粕对花鲈肝脏抗氧化指标的影响

由表8可知,与对照组相比,饲料中添加不同水平裂壶藻粕均显著提高了花鲈肝脏中超氧化物歧化酶活性(P<0.05);肝脏中过氧化氢酶和谷胱甘肽过氧化物酶活性随着裂壶藻粕添加量的增加呈先升后降的趋势,在裂壶藻粕添加量为1.6%时达到最高,并显著高于对照组(P<0.05);与对照组相比,1.6%、2.4%、3.2%裂壶藻粕添加组花鲈肝脏中丙二醛含量均显著降低(P<0.05)。
表8 裂壶藻粕对花鲈肝脏抗氧化指标的影响

Table 8 Effects of Schizochytrium meal on liver antioxidant indices of Lateolabrax maculatus

项目
Items		 裂壶藻粕添加量Schizochytrium meal additive amount/%
0(对照Control) 0.8 1.6 2.4 3.2
过氧化氢酶
CAT/(U/mg prot)		 23.89±1.15a 27.23±0.51ab 32.66±1.35c 31.08±0.92bc 28.62±2.00bc
超氧化物歧化酶
SOD/(U/mg prot)		 858.67±41.62a 966.52±31.48b 995.24±8.71b 1 044.92±38.52b 989.55±32.46b
谷胱甘肽过氧化物酶
GSH-Px/(U/mg prot)		 18.98±0.85a 22.67±0.47bc 24.29±1.59c 20.14±0.72ab 18.25±0.10a
丙二醛
MDA/(nmol/mg prot)		 0.68±0.08b 0.52±0.07ab 0.39±0.05a 0.39±0.02a 0.47±0.04a

3 讨论

3.1 裂壶藻粕对花鲈生长性能和常规成分的影响

裂壶藻是海洋微藻的一种,以往研究表明,饲料中适量添加裂壶藻可以改良饲料的适口性,促进水生生物的生长[10-11]。在大西洋鲑(Salmo salar)[12]、斑点叉尾鮰(Ictalurus punctatus)[13]和卵形鲳鲹(Trachinotus ovatus)[14]上的研究表明,饲料中添加适量的裂壶藻可以提高其生长性能。近年来,微藻的众多相关产物也广泛应用于水产饲料中,研究显示,饲料中添加裂壶藻发酵产物能够促进凡纳滨对虾(Litopenaeus vannamei)的生长且改善其肌肉品质[15],饲料中添加微藻渣同样可以显著提高凡纳滨对虾[16]和欧洲鲈(Dicentrarchus labrax L.)[17]的生长性能。裂壶藻粕作为裂壶藻的工业副产物,仍保留部分蛋白质、脂质和微量元素等,并含有丰富的DHA和藻胶多糖。本研究发现,饲料中裂壶藻粕添加量为1.6%时,显著提高了花鲈的增重率和特定生长率,这可能是因为裂壶藻粕中含有的多不饱和脂肪酸和藻胶多糖等物质改善了花鲈的生长性能;随着裂壶藻粕添加量的继续增加,花鲈的增重率和特定生长率呈现下降趋势。相似的研究结果发现,过量的微藻或其相关产物的添加会对某些水生生物的生长产生负面影响[18],饲料中裂壶藻渣添加过量会显著降低框鳞镜鲤(Cyprinus carpio var. specularis)[19]和多鳞白甲鱼(Onychostoma macrolepis)[20]的生长性能。其原因可能是微藻及相关产物中含有的单宁酸、植酸和凝集素等抗营养物质影响了鱼体的生长[21]。各裂壶藻粕添加组花鲈肌肉的粗脂肪含量均显著低于对照组,这与裂壶藻渣应用于多鳞白甲鱼所得研究结果[20]类似。这可能是因为裂壶藻副产物中含有丰富的DHA,通过促进脂质分解代谢和脂肪酸β氧化降低了鱼体肌肉中的粗脂肪含量。已有的研究发现,饲料中添加DHA或小球藻可通过调节脂质代谢的方式显著降低香鱼(Plecoglossus altivelis)[22]和黄鳝(Monopterus albus)[23]肌肉中的粗脂肪含量。

3.2 裂壶藻粕对花鲈血清生化指标的影响

甘油三酯主要在肝脏中合成,是脂质代谢中的关键组分,血清中甘油三酯的含量一定程度上可以反映机体的脂质代谢状态[24]。鱼类对糖类的利用能力较低,血清中葡萄糖含量过高会导致鱼类肝脏损伤、免疫力下降、细胞受损等[25]。以往研究发现,饲料中添加微藻可以改善尼罗罗非鱼(Oreochromis niloticus)[10]、框鳞镜鲤[19]和多鳞白甲鱼[20]的脂肪积累。本研究结果显示,饲料中添加不同水平裂壶藻粕均显著降低了花鲈血清中甘油三酯含量。这可能是由于裂壶藻粕中含有的DHA调控了花鲈的脂质代谢。已有研究表明,DHA可以通过3个途径缓解鱼体脂质沉积,一是DHA通过激活腺苷酸激活蛋白激酶(AMPK)信号通路,抑制肝脏脂肪的合成;二是DHA调节脂肪甘油三酯脂肪酶(ATGL)的活性,从而促进脂质分解;三是DHA可以诱导脂肪细胞的凋亡[26-27]。另外,藻胶多糖属于天然活性多糖,天然活性多糖有改善糖代谢、减少脂质积累的作用[28]。研究显示,饲料中添加活性多糖可以通过降低过氧化物酶体增殖物激活受体γ(PPARγ)、脂肪酸合成酶(FAS)和乙酰辅酶A羧化酶(ACC)等基因的表达,减少脂质生成,进而显著降低鱼体血清中甘油三酯和葡萄糖含量[29]。
谷丙转氨酶和谷草转氨酶是肝脏中2种重要的转氨酶,当肝脏受到损伤时,肝细胞内部的谷丙转氨酶和谷草转氨酶会释放到血液中,血清中谷丙转氨酶和谷草转氨酶的活性可以作为评估鱼类肝脏功能健康状况的重要指标[30-31]。本研究结果表明,饲料中添加0.8%、1.6%、3.2%裂壶藻粕均显著降低了花鲈血清中谷丙转氨酶和谷草转氨酶的活性,表明裂壶藻粕具有减轻肝脏损伤以及保护肝脏的作用。有研究表明,裂壶藻渣可以降低草鱼由脂肪过度积累导致的肝脏损伤[32-33]。裂壶藻粕可能是通过增强鱼体脂质代谢,减少肝脏脂质沉积,缓解肝脏损伤进而降低了血清中谷丙转氨酶和谷草转氨酶的活性。

3.3 裂壶藻粕对花鲈血清非特异性免疫力的影响

酸性磷酸酶和碱性磷酸酶是生物体免疫过程中重要的一环,酸性磷酸酶通过参与磷酸基团的代谢间接增强机体的免疫力[34],碱性磷酸酶是一种非特异性磷酸水解酶,对钙质吸收、骨骼生成具有重要的作用[35]。免疫球蛋白主要存在于血液、淋巴液、组织液等体液中,在适应性体液免疫中起重要作用,免疫球蛋白具体中和细菌毒素、激活补体、加强巨噬细胞的吞噬作用等[36]。在加州鲈(Micropterus salmoides)[37]、尼罗罗非鱼[10]和凡纳滨对虾[38]上的研究表明,饲料中添加裂壶藻均显著提高了机体的非特异性免疫力。本研究中,饲料中添加1.6%、2.4%、3.2%裂壶藻粕显著提高了花鲈血清中酸性磷酸酶的活性,且添加1.6%裂壶藻粕还显著提高了血清中免疫球蛋白M的含量,与以往研究结果类似,可能是裂壶藻粕中保留的多不饱和脂肪酸、多糖等营养物质增强了花鲈的非特异性免疫力。已有的研究表明,多不饱和脂肪酸对鱼类的免疫力有重要的调节作用[39]。另外,活性多糖能够调控鱼类免疫系统,刺激细胞因子的释放和补体的生成[40],本研究结果中各裂壶藻粕添加组花鲈血清中补体4的含量同样有上升的趋势,但并不显著。

3.4 裂壶藻粕对花鲈肝脏抗氧化能力的影响

过氧化氢酶、超氧化物歧化酶和谷胱甘肽过氧化物酶是生物抗氧化防御系统中的关键酶,抗氧化酶可以通过减少活性氧的产生,减轻氧化应激对机体的负面影响,抗氧化酶活性的变化可以一定程度上反映鱼体的抗氧化能力变化[41]。丙二醛是一种脂质过氧化产物,被认为是评价水生动物氧化应激和脂质过氧化的标志物[42]。研究显示,饲料中添加裂壶藻渣可以显著增强框鳞镜鲤[19]和多鳞白甲鱼[20]的肝脏抗氧化能力。本试验结果表明,在饲料中添加1.6%裂壶藻粕可以显著提高肝脏中过氧化氢酶、超氧化物歧化酶和谷胱甘肽过氧化物酶活性,并显著降低肝脏中丙二醛含量,该结果与在框鳞镜鲤[19]和多鳞白甲鱼[20]上的研究结果一致,可能是裂壶藻粕中含有的DHA通过调节脂质代谢的方式减少了肝脏脂质沉积,对肝脏有一定的保护作用,从而增强了花鲈肝脏的抗氧化能力[32]。当裂壶藻粕添加量超过2.4%时,花鲈肝脏中丙二醛的含量有明显升高的趋势,这表明裂壶藻粕的添加量可能过高。已有研究表明,过量的多不饱和脂肪酸摄入更容易受到氧自由基的攻击,进而发生脂质过氧化,引起鱼类肝脏中丙二醛含量的显著升高[43]。本研究中可能是过量添加裂壶藻粕引起了肝脏的抗氧化应激,体内多不饱和脂肪酸含量过多,遭受氧自由基攻击后生成大量脂质过氧化物,导致抗氧化酶活性及丙二醛含量升高。

4 结论

裂壶藻粕中含有多种营养物质,粗蛋白质含量为18.8%,粗脂肪含量为12.5%,藻胶多糖含量为11.3%,DHA含量为5.63%。饲料中添加适量裂壶藻粕能够提高花鲈的生长性能,降低肌肉粗脂肪含量,同时具有增强机体非特异性免疫力和肝脏抗氧化能力的作用,在添加量为1.6%(按干物质计)时效果最佳。

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