摘要
为研发优质的低蛋白饲料,以大口黑鲈(Micropterus salmoides)幼鱼为研究对象,设置46%(对照)、41%(LP1)和36%(LP2)3个饲料蛋白质梯度,在41%和36%蛋白饲料的基础上补充相应的晶体赖氨酸与蛋氨酸,使赖氨酸与蛋氨酸含量与对照组保持一致,分别记作LP1+AA组和LP2+AA组。进行周期为8周的饲养试验,试验结束后对大口黑鲈的生长性能和抗氧化能力进行测定。结果显示:LP1+AA组大口黑鲈的增重率、特定生长率、饲料系数和蛋白质保留率与对照组相比无显著性差异,其他各组与对照组相比发生显著性变化。随着蛋白水平降低,肌肉和全鱼中粗蛋白含量均显著降低,但LP1+AA组与对照组之间无显著性差异;粗脂肪和粗灰分含量均无显著性变化。低蛋白日粮组试验鱼的肝体比显著下降,但LP1+AA组与对照组无显著性差异。相比于对照组,LP2组和LP2+AA组试验鱼的肝脏活性氧水平和总抗氧化能力均发生显著性变化;LP1组试验鱼肝脏的总抗氧化能力显著性降低,但补充赖氨酸和蛋氨酸有明显的提高。降低大口黑鲈饲料中的蛋白水平(5%或10%)对肝脏中转氨酶的活性未产生显著影响;LP2组试验鱼血清中谷丙转氨酶活性发生显著变化,但在LP2试验组的基础上添加赖氨酸和蛋氨酸能起到改善作用,并且与对照组相比无显著性差异。结果表明,饲料粗蛋白含量从46%降低至41%并补充适量赖氨酸、蛋氨酸,对大口黑鲈幼鱼生长性能没有显著影响;但饲料粗蛋白含量降低至36%并补充赖氨酸和蛋氨酸,不能有效缓解饲料粗蛋白水平过低对生长性能造成的负面影响。
近年来,氮排放引发的水环境污染随水产养殖规模和集约化程度的不断扩大而日趋严重。水产养殖对象对饲料蛋白质的高效利用是充分发挥生长性能并降低氮排放的关键。降低日粮中蛋白水平不仅可以降低成本,还可以提高饲料中蛋白质利用效率以及降低氨氮排
大口黑鲈(Micropterus salmoides)属于典型的肉食性鱼类,对饲料中蛋白水平要求较高,其蛋白最适需求量的问题仍存在争议,提高饲料中蛋白水平能有效促进大口黑鲈生长,但会出现蛋白质利用效率下降的现
饲料原料由武汉澳华科技有限公司提供,试验饲料配方如
项目 Item | 对照 Control | LP1 | LP1+AA | LP2 | LP2+AA |
---|---|---|---|---|---|
原料/% Ingredients | |||||
鱼粉 White fish meal | 50.30 | 44.80 | 44.80 | 39.30 | 39.30 |
发酵豆粕 Fermented soybean meal | 27.60 | 24.60 | 24.60 | 21.60 | 21.60 |
豆油 Soybean oil | 7.50 | 8.00 | 8.00 | 8.50 | 8.50 |
高筋小麦粉 Wheat flour | 5.60 | 5.60 | 5.60 | 5.60 | 5.60 |
纤维素 Cellulose | — | 8.00 | 7.22 | 16.00 | 14.87 |
维生素预混料 Vitamin premix | 2.00 | 2.00 | 2.00 | 2.00 | 2.00 |
矿物质预混料 Mineral premix | 2.00 | 2.00 | 2.00 | 2.00 | 2.00 |
海藻酸钠 Sodium alginate | 1.50 | 1.50 | 1.50 | 1.50 | 1.50 |
甜菜碱 Betaine | 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 | 1.00 | 1.00 | 1.00 | 1.00 | 1.00 |
卵磷脂 Lecithin | 1.50 | 1.50 | 1.50 | 1.50 | 1.50 |
赖氨酸 Lys | — | — | 0.303 | — | 0.570 |
蛋氨酸 Met | — | — | 0.086 | — | 0.162 |
营养成分 Proximate analysis | |||||
水分/% Moisture | 8.89 | 8.88 | 8.85 | 8.52 | 8.41 |
粗蛋白/% Crude protein | 46.74 | 41.67 | 41.74 | 36.60 | 36.83 |
粗脂肪/% Crude lipid | 13.18 | 13.22 | 13.29 | 13.35 | 13.27 |
粗灰分/% Ash | 9.75 | 10.13 | 10.43 | 9.33 | 10.42 |
能量/(MJ/kg) Energy | 19.61 | 19.04 | 19.32 | 19.48 | 19.76 |
赖氨酸/% Lys | 3.153 | 2.809 | 3.112 | 2.465 | 3.035 |
蛋氨酸/% Met | 0.898 | 0.800 | 0.886 | 0.702 | 0.864 |
规格相近、健康无病的大口黑鲈500尾选购自湖北黄优源渔业发展有限公司,于华中农业大学水产养殖基地塑料养殖桶(直径120 cm,高130 cm)中进行养殖,试验过程中养殖桶是静水模式,每次喂食前使用虹吸管清理粪便、残饵等污染物,换水40%~50%,维持良好的养殖水质。暂养2周后,从中随机选取450尾初始体质量(31.52±0.62) g的试验鱼,每个养殖桶放30尾,每组饲料设置3个重复。于每天 08:00和 18:00投喂幼鱼体质量3%~6%的试验饲料。试验过程保持曝气,水温维持在25~28 ℃,氨氮为(0.25±0.12) mg/L,pH为7.2±0.2,饲养试验为期8周。
养殖周期结束后停喂24 h,取出所有试验鱼,使用MS-222溶液麻醉,记录大口黑鲈的终末体质量、存活数量以及体长等数据。每个试验组随机选取3尾作为全鱼样品放入-80 ℃冰箱冷冻保存,每个养殖桶挑选3尾试验鱼无菌操作下尾静脉采血后分离内脏和肝脏并称质量计算形体学指标,其余试验鱼采取肝脏、肌肉组织及血样,血样经过离心,分离出血清,放入-80 ℃冰箱保存用于常规营养成分、抗氧化酶及转氨酶活性的检测。
1)测定指标。试验结束对大口黑鲈增重率(WGR)、特定生长率(SGR)、饲料系数(FCR)、存活率(SR)、脏体比(VSI)、肝体比(HIS)、肥满度(CF)、蛋白质保留率(PR)、摄食量(FI)等指标进行测定,具体的计算公式参考陈乃松
2)测定方法。全鱼和肌肉中的水分测定采用105 ℃烘干法;样品中粗蛋白含量测定采用凯氏定氮法;粗脂肪测定方式使用索氏抽提法,粗灰分测定采用马弗炉550 ℃高温灼烧法;饲料总能量的测定采用氧弹法;饲料中赖氨酸和蛋氨酸的含量用氨基酸自动分析仪(Hitachi,Model 835-50,Hitachi,Tokyo,Japan)分析。使用南京建成生物工程研究所的检测试剂盒对血清和肝脏中谷草转氨酶(GOT)活性(C010-2-1)、谷丙转氨酶(GPT)活性(C009-2-1)、超氧化物歧化酶(SOD)活性(A001-3)、过氧化氢酶(CAT)活性(A007-1-1)、总抗氧化能力(T-AOC)(A015-2-1)、丙二醛(MDA)含量(A003-1)、活性氧(ROS)水平(E004-1)进行测定。样品的前期处理以及测定方法参照试剂盒说明书,大口黑鲈肝脏和血清转氨酶和抗氧化指标测定之前选取一定量的肝脏组织,加入预冷生理盐水以体积1∶9制成匀浆,在4 ℃下3 000 r/min离心机进行离心10 min,取离心后上层清液,按一定比例稀释后用于后续转氨酶和抗氧化指标活性分析。
由
指标Index | 对照Control | LP1 | LP1+AA | LP2 | LP2+AA |
---|---|---|---|---|---|
初始体质量/g IBW | 31.45±0.12 | 31.37±0.45 | 31.59±0.49 | 31.45±0.13 | 31.60±0.13 |
终末体质量/g FBW | 66.30±1.18a | 59.30±3.43bc | 63.77±4.32ab | 55.52±1.15c | 58.77±2.45bc |
增重率/% WGR | 110.86±4.37a | 90.02±11.46bc | 101.85±12.43ab | 76.51±2.94c | 85.99±8.36bc |
特定增长率/(%/d) SGR | 1.33±0.04a | 1.14±0.11bc | 1.25±0.11ab | 1.01±0.03c | 1.11±0.08bc |
摄食量/(g/尾) FI | 53.12±3.23 | 46.58±5.81 | 51.40±7.20 | 44.48±4.78 | 45.36±4.11 |
饲料系数 FCR | 1.52±0.05c | 1.65±0.02ab | 1.60±0.07bc | 1.71±0.02a | 1.67±0.04ab |
蛋白质保留率/% PR | 24.54±1.14c | 26.56±4.08b | 25.04±0.41bc | 28.08±0.92ab | 30.62±3.61a |
成活率/% SR | 100.00±0.00 | 100.00±0.00 | 100.00±0.00 | 100.00±0.00 | 100.00±0.00 |
注: 同行数值后不同的字母表示差异显著(P<0.05),相同字母或无字母表示差异不显著(P>0.05)。下表同。Note:Values within the same line with different letters are significantly different (P<0.05),same letter or no letter indicates non-significant difference(P>0.05). The same as below.
由
指标 Index | 对照 Control | LP1 | LP1+AA | LP2 | LP2+AA |
---|---|---|---|---|---|
肌肉 Muscle | |||||
水分 Moisture | 79.87±0.48 | 80.07±0.47 | 80.15±0.43 | 80.30±0.18 | 80.22±0.13 |
粗蛋白 Crude protein | 18.42±0.49a | 17.39±0.29bc | 17.91±0.39ab | 17.04±0.35c | 17.65±0.07bc |
粗脂肪 Crude lipid | 1.28±0.04 | 1.26±0.05 | 1.25±0.04 | 1.29±0.02 | 1.27±0.01 |
粗灰分 Ash | 1.26±0.01 | 1.27±0.02 | 1.23±0.04 | 1.29±0.04 | 1.25±0.02 |
全鱼 Whole fish | |||||
水分 Moisture | 74.62±0.45 | 74.80±0.44 | 74.93±0.65 | 74.58±0.83 | 74.35±0.82 |
粗蛋白 Crude protein | 17.01±0.11a | 16.16±0.40b | 16.38±0.42ab | 15.31±0.48c | 16.13±0.25b |
粗脂肪 Crude lipid | 5.12±0.05 | 5.32±0.23 | 5.30±0.06 | 5.48±0.47 | 5.56±0.12 |
粗灰分 Ash | 3.02±0.27 | 2.94±0.13 | 3.07±0.15 | 3.18±0.29 | 3.15±0.04 |
由
指标 Index | 对照 Control | LP1 | LP1+AA | LP2 | LP2+AA |
---|---|---|---|---|---|
肥满度/(g/c | 2.09±0.06 | 2.05±0.09 | 2.10±0.01 | 2.07±0.23 | 2.11±0.03 |
脏体比/% VSI | 6.41±0.18 | 6.02±0.84 | 6.27±0.07 | 6.50±0.44 | 6.32±0.03 |
肝体比/% HSI | 1.35±0.06a | 1.27±0.02b | 1.32±0.05ab | 1.28±0.01b | 1.26±0.01b |
由

图 1 低蛋白饲料中添加赖氨酸与蛋氨酸大口黑鲈幼鱼转氨酶活性的变化
Fig.1 Changes of transaminase activity of largemouth bass fed low protein diet supplemented with lysine and methionine
图中柱上字母不同表示存在显著性差异(P<0.05),相同字母或无字母表示差异不显著。Different letters on the column in the figure indicate significant difference (P<0.05),,same letter or no letter indicates non-significant difference(P>0.05).
由
指标 Index | 对照 Control | LP1 | LP1+AA | LP2 | LP2+AA |
---|---|---|---|---|---|
活性氧 ROS | 1.00±0.09c | 1.07±0.08c | 1.04±0.07c | 1.84±0.09a | 1.45±0.07b |
总抗氧化能力/(U/g) T-AOC | 0.38±0.01a | 0.32±0.02b | 0.36±0.02a | 0.30±0.01b | 0.31±0.02b |
过氧化氢酶/(U/g) CAT | 11.77±0.78a | 12.25±0.47a | 11.63±0.75a | 8.68±1.39b | 12.17±0.70a |
丙二醛/(U/g) MDA | 1.91±0.30b | 2.14±0.20ab | 1.99±0.05b | 2.41±0.11a | 2.22±0.21ab |
超氧化物歧化酶/(U/g) SOD | 32.03±1.72 | 33.32±1.88 | 30.60±2.43 | 33.21±2.15 | 31.96±3.04 |
由
指标 Index | 对照 Control | LP1 | LP1+AA | LP2 | LP2+AA |
---|---|---|---|---|---|
活性氧 ROS | 1.00±0.07c | 1.04±0.08c | 1.03±0.10c | 1.61±0.11a | 1.35±0.07b |
总抗氧化能力/(U/mL) T-AOC | 0.42±0.03 | 0.44±0.05 | 0.38±0.03 | 0.41±0.01 | 0.42±0.03 |
过氧化氢酶/(U/mL) CAT | 48.08±5.08a | 36.52±5.18b | 42.09±5.29ab | 37.22±2.12b | 40.02±7.41ab |
丙二醛/(U/mL) MDA | 34.69±5.12ab | 35.85±0.91ab | 32.39±3.05b | 39.63±3.30a | 36.75±1.60ab |
超氧化物歧化酶/(U/mL) SOD | 88.76±6.42 | 81.55±2.15 | 82.70±1.34 | 81.50±3.88 | 82.64±6.07 |
本研究结果显示降低饲料中蛋白质含量会导致大口黑鲈幼鱼生长性能和饲料利用效率显著下降,这与降低饲料蛋白质水平对宝石鲈(Scortum barcoo
本研究结果显示,饲料蛋白质含量的降低会导致全鱼粗蛋白含量下降,通过补充赖氨酸和蛋氨酸后有一定的改善作用。而程小飞
转氨酶是机体氨基酸代谢过程中的关键酶,正常生理状态下血清中的含量相对较低,但在肝、肾、心肌等组织中的含量会相对较
T-AOC的变化侧面反映机体受到外界刺激时,对自由基进行代谢以及代偿能力的高低。当机体脂质过氧化损伤时提高SOD和T-AOC的活性做出应激反
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