1 材料与方法
1.1 试验饲料
表1 饲料组成及营养水平(风干基础)Table 1 Composition and nutrient levels of diets (air-dry basis) |
| 项目 Items | 组别Groups | ||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| FM | FR | ES | SP | CP | |||||||
| 原料Ingredients | |||||||||||
| 鱼粉Fish meal | 50.00 | 45.00 | 45.00 | 45.00 | 45.00 | ||||||
| 发酵菜籽粕Fermented rapeseed meal | 9.00 | ||||||||||
| 酶解豆粕Enzymolytic soybean meal | 2.00 | 2.00 | 9.00 | 2.00 | 2.00 | ||||||
| 螺旋藻粉Spirulina powder | 5.00 | ||||||||||
| 小球藻粉Chlorella powder | 5.00 | ||||||||||
| 虾粉Shrimp meal | 5.00 | 5.00 | 5.00 | 5.00 | 5.00 | ||||||
| 小麦粉Wheat flour | 18.00 | 18.00 | 18.00 | 18.00 | 18.00 | ||||||
| α-淀粉α-starch | 9.80 | 5.60 | 7.80 | 9.60 | 9.70 | ||||||
| 谷朊粉Wheat gluten powder | 2.00 | 2.00 | 2.00 | 2.00 | 2.00 | ||||||
| 酵母粉Yeast powder | 5.00 | 5.00 | 5.00 | 5.00 | 5.00 | ||||||
| 鱼油Fish oil | 2.00 | 2.00 | 2.00 | 2.00 | 2.00 | ||||||
| 大豆油Soybean oil | 1.20 | 1.40 | 1.20 | 1.40 | 1.30 | ||||||
| 磷酸二氢钙Ca(H2PO4)2 | 2.00 | 2.00 | 2.00 | 2.00 | 2.00 | ||||||
| 卵磷脂Lecithin | 0.50 | 0.50 | 0.50 | 0.50 | 0.50 | ||||||
| 维生素预混料Vitamin premix1) | 1.00 | 1.00 | 1.00 | 1.00 | 1.00 | ||||||
| 矿物质预混料Mineral premix2) | 1.00 | 1.00 | 1.00 | 1.00 | 1.00 | ||||||
| 胆碱Choline | 0.50 | 0.50 | 0.50 | 0.50 | 0.50 | ||||||
| 合计Total | 100.00 | 100.00 | 100.00 | 100.00 | 100.00 | ||||||
| 营养水平Nutrient levels3) | |||||||||||
| 水分Moisture | 7.50 | 7.41 | 7.49 | 7.42 | 7.29 | ||||||
| 粗蛋白质CP | 41.74 | 42.82 | 41.25 | 41.27 | 42.50 | ||||||
| 粗脂肪EE | 6.42 | 6.65 | 6.77 | 6.09 | 6.58 | ||||||
| 粗灰分Ash | 11.55 | 11.77 | 11.45 | 11.88 | 11.46 | ||||||
1)维生素预混料为每千克饲料提供 The vitamin premix provided the following per kilogram of diets:VA 21 667 IU,VD 4 800 IU,VE 200 IU,VK3 25 mg,VB1 6 mg,VB2 5 mg,VB6 7.5 mg,VB12 0.03 mg,烟酰胺 nicotinamide 50mg,抗坏血酸 ascorbic acid 400 mg,泛酸钙 calcium pantothenate 25 mg,叶酸 folic acid 5 mg,生物素 biotin 1 mg,肌醇 inositol 100 mg。 3)营养水平为实测值。Nutrient levels were measured values. | |
2)矿物质预混料为每千克饲料提供 The mineral premix provided the following per kilogram of diets:MnSO4·H2O 60 mg,KI 0.1 mg,CoCl2 (1%) 3 mg,CuSO4·5H2O 15 mg,FeSO4·H2O 260 mg,ZnSO4·H2O 180 mg,Na2SeO3 (1%) 1.5 mg。 |
1.2 试验设计和饲养管理
1.3 样品采集
1.4 测定指标及方法
1.4.1 生长性能
1.4.2 肌肉常规营养成分含量
1.4.3 血清生化指标、肝脏抗氧化指标和肠道消化酶活性
1.4.4 肠道组织形态
1.4.5 肠道菌群
1.5 数据统计分析
2 结果与分析
2.1 不同植物蛋白质源部分替代鱼粉对中华鳖生长性能的影响
表2 不同植物蛋白质源部分替代鱼粉对中华鳖生长性能的影响Table 2 Effects of partial substitution of fish meal with different plant protein sources on growth performance of Chinese softshell turtle (Pelodiscus sinensis) |
| 项目 Items | 组别Groups | P值 P-value | ||||
|---|---|---|---|---|---|---|
| FM | FR | ES | SP | CP | ||
| 初始体重IBW/g | 542.33±4.91 | 542.00±4.16 | 547.67±6.69 | 545.67±3.84 | 533.00±3.00 | 0.082 |
| 终末体重FBW/g | 737.50±7.22b | 845.36±7.21a | 744.81±5.14b | 746.46±5.64b | 750.45±4.20b | <0.001 |
| 增重率WGR/% | 35.19±0.39b | 55.97±0.38a | 36.02±1.21b | 36.80±0.11b | 35.72±1.22b | <0.001 |
| 特定生长率SGR/(%/d) | 0.68±0.01b | 0.99±0.01a | 0.68±0.02b | 0.70±0.00b | 0.68±0.02b | <0.001 |
| 摄食率FR/% | 1.19±0.04ab | 1.22±0.01a | 1.13±0.03b | 1.15±0.01ab | 1.08±0.02b | 0.003 |
| 饲料系数FCR | 1.69±0.02a | 1.26±0.01b | 1.67±0.08a | 1.66±0.01a | 1.61±0.07a | <0.001 |
| 肝体比HSI/% | 3.69±0.19 | 3.19±0.28 | 3.09±0.26 | 3.38±0.23 | 3.53±0.38 | 0.156 |
| 成活率SR/% | 100.00 | 100.00 | 100.00 | 100.00 | 100.00 | |
同行数据肩标不同字母表示差异显著(P<0.05),相同字母或无字母表示差异不显著(P>0.05)。下表同。 | |
In the same row, values with different letter superscripts mean significant difference (P<0.05), while with the same letter or no letter superscripts mean no significant difference (P>0.05). The same as below. |
2.2 不同植物蛋白质源部分替代鱼粉对中华鳖肌肉常规营养成分含量的影响
表3 不同植物蛋白质源部分替代鱼粉对中华鳖肌肉常规营养成分含量的影响Table 3 Effects of partial substitution of fish meal with different plant protein sources on conventional nutrient contents in muscle of Chinese softshell turtle (Pelodiscus sinensis) |
| 项目 Items | 组别Groups | P值 P-value | ||||
|---|---|---|---|---|---|---|
| FM | FR | ES | SP | CP | ||
| 水分Moisture | 77.34±0.22a | 76.08±0.18b | 76.34±0.21ab | 75.07±0.60b | 76.00±0.48b | <0.001 |
| 粗蛋白质CP | 19.51±0.25b | 20.79±0.32a | 19.55±0.13b | 21.16±0.25a | 20.80±0.42a | <0.001 |
| 粗脂肪EE | 0.26±0.01b | 0.26±0.01b | 0.31±0.02a | 0.29±0.01a | 0.27±0.01ab | <0.001 |
| 粗灰分Ash | 1.03±0.01 | 0.99±0.06 | 1.01±0.05 | 0.99±0.03 | 1.00±0.05 | 0.625 |
2.3 不同植物蛋白质源部分替代鱼粉对中华鳖血清生化指标的影响
表4 不同植物蛋白质源部分替代鱼粉对中华鳖血清生化指标的影响Table 4 Effects of partial substitution of fish meal with different plant protein sources on serum biochemical indices of Chinese softshell turtle (Pelodiscus sinensis) |
| 项目 Items | 组别Groups | P值 P-value | ||||
|---|---|---|---|---|---|---|
| FM | FR | ES | SP | CP | ||
| 总蛋白TP/(g/L) | 17.79±0.34a | 16.71±0.79ab | 15.84±0.54b | 17.78±0.41a | 16.04±0.79ab | 0.012 |
| 白蛋白ALB/(g/L) | 13.92±1.28ab | 15.52±1.23a | 12.10±1.88b | 14.10±1.38ab | 15.01±0.86a | 0.028 |
| 总胆固醇TC/(mmol/L) | 5.63±0.05a | 6.24±0.93a | 3.79±0.56b | 5.32±0.04a | 5.22±0.07a | <0.001 |
| 甘油三酯TG/(mmol/L) | 4.44±0.40a | 1.59±0.07bc | 1.14±0.17c | 1.89±0.16b | 1.75±0.21bc | <0.001 |
| 酸性磷酸酶 ACP/(U/dL) | 5.48±0.06 | 5.52±0.43 | 5.17±0.12 | 5.71±0.14 | 5.45±0.13 | 0.215 |
| 碱性磷酸酶 AKP/(金氏单位/dL) | 16.29±1.41c | 25.55±2.17a | 20.81±1.23bc | 26.05±1.23a | 23.74±0.28ab | <0.001 |
| 谷草转氨酶AST/(U/L) | 7.45±1.18a | 6.00±0.61b | 4.99±0.75b | 5.71±0.29b | 5.31±0.78b | <0.001 |
| 谷丙转氨酶ALT/(U/L) | 1.97±0.33a | 0.69±0.09b | 0.99±0.07b | 0.68±0.07b | 0.95±0.15b | <0.001 |
2.4 不同植物蛋白质源部分替代鱼粉对中华鳖肝脏抗氧化指标的影响
表5 不同植物蛋白质源部分替代鱼粉对中华鳖肝脏抗氧化指标的影响Table 5 Effects of partial substitution of fish meal with different plant protein sources on liver antioxidant indices of Chinese softshell turtle (Pelodiscus sinensis) |
| 项目 Items | 组别Groups | P值 P-value | ||||
|---|---|---|---|---|---|---|
| FM | FR | ES | SP | CP | ||
| 总抗氧化能力 T-AOC/(U/g prot) | 1.28±0.22b | 2.03±0.22a | 0.98±0.12b | 2.06±0.20a | 1.21±0.14b | <0.001 |
| 过氧化氢酶 CAT/(U/g prot) | 817.58±57.99b | 844.08±85.96ab | 1 031.20±26.54a | 802.88±111.10b | 998.36±72.38ab | <0.001 |
| 超氧化物歧化酶 SOD/(U/mg prot) | 58.64±0.24b | 63.49±1.34ab | 66.52±2.53a | 67.41±1.48a | 51.84±2.71c | <0.001 |
| 丙二醛 MDA/(nmol/mg prot) | 2.98±0.53a | 1.64±0.03b | 2.98±0.32a | 2.57±0.42a | 2.74±0.22a | <0.001 |
2.5 不同植物蛋白质源部分替代鱼粉对中华鳖肠道消化酶活性的影响
表6 不同植物蛋白质源部分替代鱼粉对中华鳖肠道消化酶活性的影响Table 6 Effects of partial substitution of fish meal with different plant protein sources on intestinal digestive enzyme activity of Chinese softshell turtle (Pelodiscus sinensis) U/mg prot |
| 项目 Items | 组别Groups | P值 P-value | ||||
|---|---|---|---|---|---|---|
| FM | FR | ES | SP | CP | ||
| 淀粉酶Amylase | 0.11±0.01b | 0.12±0.01b | 0.04±0.01c | 0.11±0.01b | 0.19±0.04a | <0.001 |
| 脂肪酶Lipase | 17.12±1.32bc | 11.47±0.57c | 22.06±2.03b | 19.47±3.37bc | 30.71±3.72a | <0.001 |
| 胰蛋白酶Trypsin | 585.35±16.42b | 595.41±78.37b | 624.62±61.07b | 596.98±43.32b | 1 265.50±198.70a | <0.001 |
2.6 不同植物蛋白质源部分替代鱼粉对中华鳖肠道组织形态的影响
图1 中华鳖肠道组织切片VH:绒毛高度 villus height;CD:隐窝深度 crypt depth;MT:肌层厚度 muscular layer thickness。 Fig.1 Sections of intestinal tissues of Chinese softshell turtle (Pelodiscus sinensis) (200×) |
表7 不同植物蛋白质源部分替代鱼粉对中华鳖肠道组织形态的影响Table 7 Effects of partial substitution of fish meal with different plant protein sources on intestinal tissue morphology of Chinese softshell turtle (Pelodiscus sinensis) |
| 项目 Items | 组别Groups | P值 P-value | ||||
|---|---|---|---|---|---|---|
| FM | FR | ES | SP | CP | ||
| 绒毛高度Villus height | 856.82±33.25a | 579.49±14.75b | 643.05±23.93b | 819.06±27.47a | 668.66±42.55b | <0.001 |
| 隐窝深度Crypt depth | 76.91±1.01a | 61.55±1.36c | 69.49±4.01b | 68.97±1.98b | 74.31±2.70ab | <0.001 |
| 肌层厚度 Muscular layer thickness | 374.41±6.80a | 350.45±8.65b | 322.45±7.77c | 362.57±2.45ab | 335.00±9.08c | <0.001 |
2.7 不同植物蛋白质源部分替代鱼粉对中华鳖肠道菌群的影响
2.7.1 α多样性分析
表8 不同植物蛋白质源部分替代鱼粉对中华鳖肠道菌群α多样性的影响Table 8 Effects of partial substitution of fish meal with different plant protein sources on α diversity of intestinal microbiota of Chinese softshell turtle (Pelodiscus sinensis) |
| 项目 Items | 组别Groups | P值 P-value | ||||
|---|---|---|---|---|---|---|
| FM | FR | ES | SP | CP | ||
| Shannon指数 Shannon index | 4.58±2.01 | 3.55±1.67 | 3.79±1.72 | 3.87±0.75 | 2.28±1.82 | 0.105 |
| Simpson指数 Simpson index | 0.81±0.19 | 0.74±0.20 | 0.85±0.18 | 0.86±0.08 | 0.65±0.17 | 0.098 |
| Chao1指数Chao1 index | 485.40±83.63ab | 460.64±40.03ab | 359.71±6.60b | 500.38±40.60a | 441.95±103.79ab | 0.021 |
| Sobs指数Sobs index | 452.00±95.41a | 397.00±63.49abc | 307.67±15.53bc | 423.67±55.42ab | 302.33±48.81c | 0.006 |
| Ace指数Ace index | 489.99±80.31a | 460.32±31.77ab | 377.98±14.73bc | 484.24±47.61a | 359.05±49.08c | 0.002 |
| Goods覆盖度 Goods coverage | 0.99±0.00 | 0.99±0.00 | 0.99±0.00 | 0.99±0.00 | 0.99±0.00 | 0.987 |
2.7.2 β多样性分析
2.7.3 物种组成分析
表9 不同植物蛋白质源部分替代鱼粉对中华鳖肠道菌群相对丰度的影响Table 9 Effects of partial substitution of fish meal with different plant protein sources on relative abundance of intestinal microbiota of Chinese softshell turtle (Pelodiscus sinensis) |
| 项目 Items | 组别Groups | P值 P-value | |||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| FM | FR | ES | SP | CP | |||||||||
| 门水平Phylum level | |||||||||||||
| 芽孢杆菌门Bacillota | 31.71±12.15 | 13.89±8.93 | 21.64±7.27 | 10.53±5.52 | 32.82±17.94 | 0.065 | |||||||
| 弯曲杆菌门 Campylobacterota | 2.92±2.48b | 8.40±5.23b | 12.03±6.38b | 44.47±17.28a | 44.40±28.63a | <0.001 | |||||||
| 假单胞菌门 Pseudomonadota | 27.92±11.07 | 7.62±4.21 | 28.87±11.68 | 25.72±6.51 | 13.36±10.56 | 0.058 | |||||||
| 拟杆菌门Bacteroidota | 24.24±12.93a | 2.49±1.51b | 5.12±3.30b | 2.67±1.57b | 0.57±0.33b | <0.001 | |||||||
| 热脱硫杆菌门 Thermodesulfobacteriota | 0.92±0.39b | 0.28±0.14b | 45.49±21.39a | 6.18±2.72b | 0.14±0.02b | <0.001 | |||||||
| 属水平Genus | |||||||||||||
| 螺杆菌属Helicobacter | 19.57±3.17 | 29.30±4.07 | 10.36±8.87 | 39.82±13.72 | 36.75±14.64 | 0.072 | |||||||
| 劳森菌属Lawsonia | 0.43±0.63b | 0.13±0.06b | 36.27±4.27a | 2.63±5.25b | 0.05±0.02b | <0.001 | |||||||
| 芽孢杆菌属Bacillus | 1.90±1.99 | 0.12±0.14 | 0.11±0.14 | 0.52±0.78 | 30.60±20.48 | 0.171 | |||||||
| 软质体属Malacoplasma | 2.61±2.11 | 1.18±1.31 | 3.82±2.91 | 14.95±10.75 | 2.34±2.38 | 0.053 | |||||||

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