Comparison and evaluation of nutrition composition in muscle of grass carp Ctenopharyngodon idellus fed with broad bean and common compound feed
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摘要:
对投喂蚕豆和普通配合饲料的2组草鱼(Ctenopharyngodon idellus)肌肉中的胶原蛋白、钙离子(Ca2+)、氨基酸和脂肪酸含量进行了比较分析，并对其营养价值与食用价值进行了评价。结果显示，投喂蚕豆草鱼肌肉中胶原蛋白和Ca2+的质量分数分别为(0.149±0.011)%和(0.036±0.011)%，比投喂普通配合饲料的草鱼分别提高了36.7%和154%。投喂蚕豆草鱼肌肉中氨基酸总量为(16.802±1.823)%，投喂普通配合饲料草鱼氨基酸总量为(18.444±0.850)%；投喂蚕豆草鱼肌肉中4种鲜味氨基酸[天冬氨酸(Asp)、谷氨酸(Glu)、甘氨酸(Gyl)和丙氨酸(Ala)]总质量分数为(6.613±0.771)%，低于投喂普通配合饲料草鱼。投喂蚕豆草鱼和投喂普通配合饲料草鱼肌肉中不饱和脂肪酸(UFA)总量分别占脂肪酸总量的73.857%和76.334%，其中单不饱和脂肪酸(MUFA)分别占脂肪总量的(55.598±2.403)%和(33.832±2.755)%。结果表明，投喂蚕豆草鱼肌肉中胶原蛋白和Ca2+含量比投喂普通配合饲料的草鱼有显著提高，但其鲜味程度、氨基酸以及不饱和脂肪酸含量比投喂普通配合饲料草鱼低。
Abstract:To study the nutrition and food value of grass carp (Ctenopharyngodon idellus), we fed the fish with broad bean and common compound feed to compare and analyze the contents of collagen, Ca2+, amino acids and fatty acids in their muscle. The results reveal that compared with group of common compound feed, contents of collagen and Ca2+ in group of broad bean are (0.149±0.011)% and (0.036±0.011)%, respectively, which increase 36.7% and 154%, respectively. The total content of amino acids in groups of broad bean and common compound feed are (16.802±1.823)% and (18.444±0.850)%, respectively. The total 4 kinds of delicious amino acids account for (6.613±0.771)% in group of broad bean, which is lower than those in group of common compound feed. The content of unsaturated fatty acids in groups of broad bean and common compound feed account for 73.857% and 76.334% of total fatty acids, respectively, and monunsaturated fatty acids account for(55.598±2.403)% and (33.832±2.755)% of total fat, respectively. It shows that the contents of collagen and Ca2+ of grass carp fed with broad bean are higher than those fed with common compound feed, while the flavor, contents of amino acids and unsaturated fatty acids of grass carp fed with broad bean are inferior to those fed with common compound feed.
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在饲料中添加增强鱼体免疫力和抵抗力的添加剂，逐渐成为控制鱼病和健康养殖的重要措施。壳聚糖是甲壳素脱乙酰基的产物，饲料中添加壳聚糖可通过提高动物的免疫功能和抗病力促进动物的生长，并且具有良好的生理活性和生物相容性，能被生物降解，且降解产物不会对环境造成污染。目前的壳聚糖生产大多以虾、蟹壳为材料，但用这些原料来生产壳聚糖存在着一定的局限性，如含量低(通常约占干重6%~20%)，虾、蟹壳中含有大量的石灰质，迫使在甲壳素生产过程中大量地使用盐酸而造成甲壳素酸降解，降低产品质量，并污染环境等。昆虫体壳甲壳素的含量最高，体壁灰分含量少，较虾、蟹壳易于提取甲壳素及减少甲壳素分子链酸降解，相对容易生产[1]。笔者以往的研究表明，昆虫源壳聚糖对蛋鸡、肉鸡的生产性能有促进作用，其生物活性明显优于虾、蟹壳源壳聚糖[2-3]。昆虫源壳聚糖在鱼类方面的研究还鲜见报道。此试验旨在鲫鱼(Carassius auratus)基础饲料中添加不同水平的昆虫源壳聚糖，探讨其在饲料中的适宜添加水平，为昆虫源壳聚糖的开发应用提供依据。
1. 材料与方法
1.1 试验设计和试验鱼
采用单因子随机区组设计。试验用鱼高背鲫购于市场，共计225尾，体重25.2±1.7 g。试验鱼在水族箱内驯养一周后随机分为5个处理组，每处理组3个重复，每组15尾。各个处理组的鲫鱼饲喂的饲料中分别添加0%、0.25%、0.50%、0.75%和1.00%的昆虫源壳聚糖，试验期为8周。
1.2 试验饲料
试验基础饲料的组成和主要营养指标见表 1。
表 1 基础饲料的组成及主要营养指标Table 1. Composition and nutrient levels of basal feed% 原料名称
feed stuff在饲料中的比例
percent in feed主要营养指标
nutrient parameters营养水平
nutrient level鱼粉 fish meal 12 粗蛋白 crude protein 30.25 豆粕 soybean meal 30 粗脂肪 crude fat 4.85 棉籽粕 cotton seed meal 11 粗灰分 crude ash 12.30 面粉 flour 15 赖氨酸 Lys 1.63 米糠 rice bran 20 蛋氨酸 Met 0.49 玉米粉 corn 10 磷酸氢钙 calcium phosphate dibasic 1 矿物质及维生素预混料 mineral and vitamin premix 1 试验饲料为基础饲料中添加0%、0.25%、0.50%、0.75%和1.00%的昆虫源壳聚糖，混合均匀后压制成2~3 mm颗粒饲料烘干备用。昆虫源壳聚糖以油葫芦(Cryllus testaceus)为原料，按照王敦等[4]的方法制备。
1.3 饲养管理
试验在水族箱中进行，其规格为120 cm×50 cm×60 cm，配有加热棒和充氧机，保持水温24~27℃，pH 7.2，溶氧5~8 mg · L-1，驯化饲养1周后饲喂试验饲料。试验鱼每天喂4次(8: 30，11: 30，14: 00，17: 00)，日投饵率2%~4%，根据鱼的摄食情况适当增减。每天记录摄食量和水温，并清除水箱内粪便和污物，定期换水。
1.4 测定指标及方法
在试验正式开始及结束前鱼饥饿24 h，给每组鱼称重。按下列公式计算相对增重率和饵料系数。
相对增重率(%) = (试验结束鱼体重－试验开始鱼体重) /试验开始鱼体重×100%；
饵料系数=TF/ (WF－WI)；
式中WF为试验终末每个处理组鱼总重；WI为初始每个处理组鱼总重；TF为每个重复摄食量。
1.5 统计分析
所有数据采用Excel软件进数据整理，用SPSS 11.5软件的One-Way ANOVA模块进行方差分析，用Turkey法进行多重比较，显著水平为P<0.05。
2. 结果与讨论
在鲫鱼饲料中添加不同水平的昆虫源壳聚糖后生长性能的变化情况见表 2。
表 2 昆虫源壳聚糖添加水平对鲫鱼生长性能的影响Table 2. Effect of chitosan from insect resource on growth performance of crucian carp壳聚糖添加水平/%
chitosan level饲料投喂量/g
feed consumption per fish相对增重率/%
relative growth rate饵料系数
feed coefficient0 24.92±1.02a 49.40±3.52a 2.01±0.10a 0.25 23.66±1.15a 55.78±2.44ab 1.69±0.13b 0.50 24.32±1.58a 63.49±3.75b 1.52±0.07b 0.75 23.70±1.32a 59.52±3.05b 1.58±0.12b 1.00 25.70±1.22a 57.71±3.25b 1.76±0.15b 注：同列数据标有相同字母者表示差异不显著(P>0.05)；标有不同字母者表示差异显著(P < 0.05)
Note：Means within row with same superscripts were not significant (P>0.05)；means within row with different superscripts were significant (P < 0.05)可以看出，添加昆虫源壳聚糖后各处理组的相对增重率均高于未添加组，其中0.50%、0.75%和1.00%处理组与对照组存在显著的差异(P < 0.05)，0.25%添加组与未添加组无显著差异(P>0.05)，0.50%处理组最高；添加昆虫源壳聚糖后的饵料系数均显著的降低(P < 0.05)，以0.50%的添加水平组最佳。其他学者在罗非鱼(Tilapia)[5]、草鱼(Ctenopharyngodon idellus)[6]、异育银鲫(Carassius auratus gibelio)[7]等鱼类饲料添加普通壳聚糖后对生长性能的影响也有相同的报道。
试验中，昆虫源壳聚糖的促生长作用并不随着壳聚糖添加量的增加而不断增强。于东祥等[8]对真鲷(Pagrosomus major)的研究有类似报道，饲料中添加0.5%和1.0%的甲壳胺制剂可以提高真鲷的相对增重率，并且0.5%的添加效果要比1.0%的效果更好。SHIAU和YU[9]的研究中也发现，罗非鱼的增重率随壳聚糖添加量(2%、5%、10%)的增加而降低，10%的壳聚糖会抑制罗非鱼的生长和饲料转化率。可见，过量添加壳聚糖对鱼的生长会有抑制作用。
通过对昆虫源壳聚糖的添加水平与相对增重率和饵料系数间的关系进行统计分析，发现存在着显著的二次曲线关系(P < 0.01)(图 1、图 2)。通过求导计算最佳相对增重率和饵料系数时的添加水平为0.63%和0.57%，平均为0.60%。这与其他学者在不同鱼类中添加虾蟹壳壳聚糖的研究结果接近，大多数学者认为虾蟹壳壳聚糖在罗非鱼[5]、真鲷幼鱼[8]、草鱼[6, 10]、花鲈(Lateolabrax japonicus)[11]、异育银鲫[12]、三角鲂(Megalobrama terminalis)[13]饲料中的适宜添加水平为0.5%，也有认为罗非鱼为0.75%[7]，甚至1.0%[14]。这可能是不同研究中的试验鱼种类、基础饲料组成、试验中壳聚糖添加梯度大小及数量等方面的差异导致的。文章通过分析壳聚糖添加水平与相对增重率和饵料系数间的关系得到的最佳添加水平，而其他学者一般是比较了不同处理组间的差异情况得出的结论，未研究壳聚糖水平与生长性能指标间的关系后通过计算确定最佳水平，这可能是与他人报道存在一定差异的主要原因。此外，昆虫源壳聚糖和虾蟹壳壳聚糖对鱼类的促生长作用是否存在差异以及机理还有待于进一步研究。
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图 1 昆虫源壳聚糖添加水平与鲫鱼相对增重率间的关系Figure 1. Relationship between levels of chitosan from insect resource and relative growth rate of crucian carp![]()
图 2 昆虫源壳聚糖添加水平与鲫鱼饵料系数间的关系Figure 2. Relationship between levels of chitosan from insect resource and feed coefficient of crucian carp3. 小结
从文中的试验结果来看，饲料中添加0.50%昆虫源壳聚糖组的生长性能最好，结合昆虫源壳聚糖水平与相对增重率和饵料系数的回归关系计算出最佳添加水平为0.60%的结果，再考虑其他学者在不同鱼类饲料中添加虾蟹壳壳聚糖的适宜水平为0.50%的情况，笔者认为，为了取得较好的养殖效果，在鲫鱼饲料中添加0.50%~0.60%昆虫源壳聚糖较为适宜。
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表 1 投喂蚕豆和普通配合饲料草鱼肌肉中胶原蛋白和Ca2+质量分数(X±SD)
Table 1 Contents of collagen and Ca2+ in muscle of grass carp fed with broad bean and common compound feed
% 组别
groupw(胶原蛋白)
collagen contentw(Ca2+)
Ca2+ contentw(水分)
moisture content脆肉鲩crisp grass carp 0.149±0.011* 0.036±0.011** 74.12±3.82 普通草鱼common grass carp 0.109±0.008 0.023±0.003 77.45±5.78 注： *. 差异显著(P < 0.05)；* *. 差异极显著(P < 0.01)，后表同此
Note： *. significant difference (P < 0.05)；* *. very significant difference (P < 0.01); the same case in the following tables.
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表 2 投喂蚕豆和普通配合饲料草鱼的氨基酸组成及含量(X±SD)
Table 2 Composition and content of amino acids in muscle of grass carp fed with broad bean and common compound feed
% 成分
ingredient脆肉鲩
crisp grass carp普通草鱼
common grass carp成分
ingredient脆肉鲩
crisp grass carp普通草鱼
common grass carp组氨酸①His 0.403±0.044 0.452±0.030* 谷氨酸④Glu 2.793±0.300 3.130±0.147* 精氨酸①Arg 1.007±0.122 1.126±0.066 甘氨酸④Gly 0.817±0.142 0.960±0.066* 胱氨酸①Cys 0.120±0.077 0.134±0.086 天冬氨酸④Asp 1.820±0.193 2.000±0.078 缬氨酸②Val 0.833±0.075 0.940±0.061* 丙氨酸④Ala 1.183±0.136 1.277±0.049 蛋氨酸②Met 0.397±0.110 0.437±0.012 总量TAA 16.802±1.823 18.444±0.850 苏氨酸②Thr 0.790±0.078 0.870±0.046* 必需氨基酸总量TEAA 6.709±0.732 7.564±0.329 苯丙氨酸②Phe 0.763±0.084 0.840±0.020* 半必需氨基酸总量THEAA 1.530±0.166 1.712±0.096 异亮氨酸②Ile 0.773±0.067 0.883±0.055* 非必需氨基酸量TNEAA 1.950±0.154 1.803±0.085 亮氨酸②Leu 1.440±0.139 1.607±0.059* 呈鲜味氨基酸总量TDAA 6.613±0.771 7.367±0.340 赖氨酸②Lys 1.713±0.179 1.987±0.076* TEAA/TAA 0.399±0.044 0.410±0.018 脯氨酸③Pro 0.607±0.091 0.350±0.026* THEAA/TAA 0.091±0.010 0.093±0.005 酪氨酸③Tyr 0.587±0.050 0.637±0.025* TEAA/TNEAA 0.663±0.072 0.695±0.030 丝氨酸③Ser 0.756±0.081 0.816±0.035* 注： ①. 半必需氨基酸；②. 必需氨基酸；③. 非必需氨基酸；④. 鲜味氨基酸
Note： ①. semi-essential amino acid；②. essential amino acid；③. non-essential amino acid；④. delicious amino acid
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表 3 投喂蚕豆草鱼与其他经济鱼类肌肉必需氨基酸模式、AAS、CS和EAAI
Table 3 Comparison of pattern of essential amino acids, AAS, CS and EAAI between grass carp and some other commercial fishes fed with broad bean
必需氨基酸
essential amino acid异亮
Ile亮
Leu赖
Lys苏
Thr缬
Val蛋+胱
Met+Cys苯丙+酪
Phe+TyrEAAI 氨基酸质量分数/mg·g-1 content of amino acid 脆肉鲩crisp grass carp 172 320 380 175 185 88 300 50.82 普通草鱼common grass carp 196 357 441 193 209 97 328 56.94 FAO/WHO模式/mg·g-1 pro FAO/WHO pattern 250 440 340 250 310 220 380 鸡蛋蛋白模式/mg·g-1pro egg protein pattern 331 534 441 292 411 386 565 氨基酸评分AAS 脆肉鲩crisp grass carp 0.69 0.73 1.12 0.70 0.60 0.40 0.79 普通草鱼common grass carp 0.78 0.81 1.30 0.77 0.62 0.44 0.86 化学评分CS 脆肉鲩crisp grass carp 0.52 0.60 0.86 0.60 0.45 0.23 0.53 普通草鱼common grass carp 0.60 0.67 1.00 0.66 0.51 0.25 0.58
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表 4 投喂蚕豆草鱼与其他经济鱼类的A/E值比较
Table 4 Comparison of A/E ratio between grass carp and some other commercial fishes fed with broad bean
必需氨基酸
essential amino acidA/E比值 A/E ratio 脆肉鲩
crisp grass carp普通草鱼
common grass carp大西洋鲑[19]
S.salar日本鳗鲡[20]
A.japonica赖氨酸Lys 25.56 26.25 23.82 22.53 苏氨酸Thr 11.79 11.49 12.74 11.23 缬氨酸Val 12.43 12.42 13.16 13.37 亮氨酸Leu 21.49 21.23 20.91 19.96 异亮氨酸Ile 11.53 11.66 12.05 12.45 苯丙氨酸Phe 11.39 11.10 9.70 10.62 蛋氨酸Met 5.90 5.77 7.62 9.89
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表 5 投喂蚕豆和普通配合饲料草鱼肌肉的脂肪酸组成及含量(X±SD)
Table 5 Composition and content of fatty acids in muscle of grass carp fed with broad bean and common compound feed
% 脂肪酸
fatty acids脆肉鲩
crisp grass carp普通草鱼
common grass carpC12：0 0.037±0.014 0.031±0.004 C14：0 0.943±0.076 0.827±0.131* C15：0 0.173±0.021 0.355±0.021** C16：0 19.570±1.364 17.527±0.850* C17：0 0.273±0.032 0.623±0.012** C18：0 4.707±0.260 3.570±0.226* C19：0 0.253±0.038 0.510±0.087** C20：0 0.187±0.021 0.223±0.032 ΣSFA 26.143±2.015 23.666±2.726* C14：1 0.069±0.003 0.090±0.005 C16：1 8.271±0.367 4.132±0.784** C17：1 0.240±0.044 0.300±0.010 C18：1 45.250±1.723 28.430±1.699** C20：1 1.680±0.255 0.823±0.159** C22：1 0.088±0.011 0.059±0.008 ΣMUFA 55.598±2.403 33.832±2.755** C16：2 0.123±0.025 0.257±0.023* C18：2 n-6 11.837±0.664 25.583±3.121** C18：3 n-3 1.177±0.287 8.997±1.126** C18：4 n-3 0.098±0.013 0.092±0.018 C20：2 0.977±0.075 1.087±0.084 C20：3 n-3 1.350±0.075 1.910±0.079 C20：4 n-6 1.237±0.298 1.770±0.141 C20：5 n-3 0.170±0.069 0.473±0.021** C22：3 n-3 0.173±0.055 0.176±0.088 C22：4 n-3 0.460±0.090 0.337±0.075 C22：5 n-3 0.107±0.024 0.473±0.047** C22：6 n-3 0.550±0.331 1.347±0.104** ΣPUFA 18.259±2.006 42.502±4.927 EPA+DHA 0.720±0.355 1.820±0.151** Σ n-3 4.085±0.854 13.805±1.483** Σ n-6 13.074±1.052 27.353±3.337** n-3/n-6 0.312±0.063 0.505±0.054* ΣHUFA 2.622±0.942 4.492±0.555 总量total content 100.000±6.424 100.000±10.408
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