1 材料与方法
1.1 试验设计
表1 基础饲粮组成及营养水平(风干基础)Table 1 Composition and nutrient levels of the basal diet (air-dry basis)% |
| 原料 Ingredients | 含量 Content | 营养水平 Nutrient levels2) | 含量 Content | |
|---|---|---|---|---|
| 玉米 Corn | 5.0 | 消化能 DE/(MJ/kg) | 10.23 | |
| 豆粕 Soybean meal | 8.0 | 干物质 DM | 89.82 | |
| 大麦 Barley | 6.0 | 粗蛋白质 CP | 16.12 | |
| 麸皮 Wheat bran | 15.0 | 粗脂肪 EE | 2.80 | |
| 玉米胚芽粕 Corn germ meal | 16.0 | 粗纤维 CF | 17.38 | |
| 玉米皮 Corn husk | 17.0 | 中性洗涤纤维 NDF | 38.74 | |
| 苜蓿草粉 Alfalfa meal | 15.0 | 酸性洗涤纤维 ADF | 23.08 | |
| 豆秸粉 Soybean straw powder | 7.0 | 酸性洗涤木质素 ADL | 6.29 | |
| 稻壳粉 Rice hull powder | 8.0 | 粗灰分 Ash | 9.03 | |
| 磷酸氢钙 CaHPO4 | 1.5 | 钙 Ca | 0.95 | |
| 氯化钠 NaCl | 0.5 | 总磷 TP | 0.45 | |
| 预混料 Premix1) | 1.0 | 赖氨酸 Lys | 0.60 | |
| 合计 Total | 100.0 | 蛋氨酸+半胱氨酸 Met+Cys | 0.65 |
1)预混料为每千克饲粮提供 Premix provided the following per kg of the diet: VA 8 000 IU,VD3 1 500 IU,VE 45 mg,VK3 2.0 mg,VB1 1.0 mg,VB2 3.0 mg,VB6 1.5 mg,烟酸 nicotinic acid 30 mg,泛酸 pantothenic acid 50 mg,叶酸 folic acid 0.5 mg,氯化胆碱 choline chloride 100 mg,Fe 50 mg,Cu 10 mg,Zn 50 mg,Mn 10 mg,I 0.5 mg,Se 0.05 mg,赖氨酸 Lys 1.5 g,蛋氨酸 Met 0.5 g,其余由麦饭石载体补足 the rest was Maifan stone carrier complement. 2)消化能为计算值,其余为实测值。DE was a calculated value, while the others were measured values. |
1.2 样品采集与制备
1.3 测定指标和方法
1.3.1 生长性能
1.3.2 屠宰性能
1.3.3 十二指肠抗氧化指标
1.3.4 空肠组织形态结构
1.3.5 回肠黏膜免疫指标
1.3.6 盲肠微生物区系
1.4 数据处理
2 结果
2.1 丁酸梭菌、葡萄糖氧化酶和迷迭香提取物组方对肉兔生长性能的影响
表2 丁酸梭菌、葡萄糖氧化酶和迷迭香提取物组方对肉兔生长性能的影响Table 2 Effects of Clostridium butyricum, glucose oxidase and rosemary extract formulas on growth performance of meat rabbits |
| 项目 Items | 组别 Groups | 均方根 误差 RMSE | P值 P-value | ||||
|---|---|---|---|---|---|---|---|
| Ⅰ (对照 Control) | Ⅱ | Ⅲ | Ⅳ | Ⅴ | |||
| 初始体重 IBW/g | 883.1 | 833.6 | 846.0 | 887.9 | 869.9 | 51.813 4 | 0.500 8 |
| 终末体重 FBW/g | 2 161.3b | 2 122.5b | 2 163.8b | 2 155.0b | 2 244.2a | 86.777 4 | 0.043 1 |
| 平均日增重 ADG/(g/d) | 42.60b | 42.96b | 43.93ab | 42.24b | 45.81a | 1.695 8 | 0.026 9 |
| 平均日采食量 ADFI/(g/d) | 167.08 | 168.92 | 168.33 | 170.50 | 169.00 | 6.320 4 | 0.820 4 |
| 料重比 F/G | 3.94ab | 3.94ab | 3.85ab | 4.05a | 3.70b | 0.274 1 | 0.049 8 |
| 成活率 Survival rate/% | 86.00b | 92.00ab | 93.00ab | 92.00ab | 97.00a | 7.272 5 | 0.030 9 |
| 腹泻率 Diarrhea rate/% | 17.00a | 8.00b | 9.00b | 7.00b | 4.00b | 8.576 5 | 0.021 6 |
同行数据肩标不同小写字母表示差异显著(P<0.05),相同或无字母表示差异不显著(P>0.05)。下表同。 | |
In the same row, values with different small letter superscripts mean significant difference (P<0.05), while with the same or no letter superscripts mean no significant difference (P>0.05). The same as below. |
2.2 丁酸梭菌、葡萄糖氧化酶和迷迭香提取物组方对肉兔屠宰性能的影响
表3 丁酸梭菌、葡萄糖氧化酶和迷迭香提取物组方对肉兔屠宰性能的影响Table 3 Effects of Clostridium butyricum, glucose oxidase and rosemary extract formulas on slaughter performance of meat rabbits |
| 项目 Items | 组别 Groups | 均方根 误差 RMSE | P值 P-value | ||||
|---|---|---|---|---|---|---|---|
| Ⅰ (对照 Control) | Ⅱ | Ⅲ | Ⅳ | Ⅴ | |||
| 宰前活体重 Pre-slaughter body weight/g | 2 141.0b | 2 142.4b | 2 191.3b | 2 160.9b | 2 282.5a | 165.981 5 | 0.042 5 |
| 欧洲屠宰率 European slaughter ratio/% | 55.19 | 55.91 | 55.49 | 57.48 | 56.80 | 2.616 1 | 0.276 7 |
| 半净膛屠宰率 Semi clean slaughter ratio/% | 49.99 | 49.95 | 49.84 | 51.26 | 50.42 | 2.070 6 | 0.528 0 |
| 全净膛屠宰率 Full clean slaughter ratio/% | 44.86 | 44.83 | 44.68 | 46.04 | 45.36 | 2.169 8 | 0.621 8 |
| 心脏相对重量 Heart relative weight/(g/kg) | 3.53 | 3.00 | 3.56 | 3.41 | 3.55 | 0.563 8 | 0.153 3 |
| 肝脏相对重量 Liver relative weight/(g/kg) | 42.21 | 40.82 | 41.19 | 40.31 | 39.59 | 4.221 2 | 0.707 6 |
| 肾脏相对重量 Kidney relative weight/(g/kg) | 8.04 | 7.81 | 8.43 | 7.95 | 8.29 | 0.765 6 | 0.358 3 |
2.3 丁酸梭菌、葡萄糖氧化酶和迷迭香提取物组方对肉兔十二指肠抗氧化指标的影响
表4 丁酸梭菌、葡萄糖氧化酶和迷迭香提取物组方对肉兔十二指肠抗氧化指标的影响Table 4 Effects of Clostridium butyricum, glucose oxidase and rosemary extract formulas on duodenal antioxidant indices of meat rabbits |
| 项目 Items | 组别 Groups | 均方根 误差 RMSE | P值 P-value | ||||
|---|---|---|---|---|---|---|---|
| Ⅰ (对照 Control) | Ⅱ | Ⅲ | Ⅳ | Ⅴ | |||
| 谷胱甘肽过氧化物酶 GSH-Px/(U/mg prot) | 13.78a | 11.09bc | 13.48ab | 10.78c | 10.94bc | 2.434 4 | 0.032 6 |
| 超氧化物歧化酶 SOD/(U/mg prot) | 272.27 | 305.10 | 302.64 | 299.08 | 320.31 | 52.714 1 | 0.489 1 |
| 丙二醛 MDA/(nmol/mg prot) | 0.51c | 0.47c | 0.56bc | 0.79a | 0.72ab | 0.172 4 | 0.002 1 |
| 总抗氧化能力 T-AOC/(mmol/g prot) | 0.56c | 1.93b | 3.03ab | 2.73ab | 3.37a | 1.182 5 | 0.000 3 |
2.4 丁酸梭菌、葡萄糖氧化酶和迷迭香提取物组方对肉兔空肠组织形态结构的影响
表5 丁酸梭菌、葡萄糖氧化酶和迷迭香提取物组方对肉兔空肠组织形态结构的影响Table 5 Effects of Clostridium butyricum, glucose oxidase and rosemary extract formulas on jejunal histological structure of meat rabbits |
| 项目 Items | 组别 Groups | 均方根 误差 RMSE | P值 P-value | ||||
|---|---|---|---|---|---|---|---|
| Ⅰ (对照 Control) | Ⅱ | Ⅲ | Ⅳ | Ⅴ | |||
| 绒毛高度 Villus height/μm | 625.36b | 806.71ab | 825.50ab | 804.57ab | 914.70a | 187.245 9 | 0.048 2 |
| 隐窝深度 Crypt depth/μm | 119.65 | 113.98 | 124.95 | 114.91 | 124.12 | 20.184 7 | 0.733 4 |
| 绒毛宽度 Villus width/μm | 107.48 | 124.14 | 128.89 | 114.47 | 140.45 | 26.117 4 | 0.129 3 |
| 黏膜层厚度 Mucosal layer thickness/μm | 787.01 | 955.70 | 979.06 | 951.78 | 1 069.63 | 196.691 3 | 0.094 2 |
| 肌层厚度 Muscle layer thickness/μm | 169.92 | 142.93 | 174.27 | 169.93 | 140.13 | 41.424 6 | 0.302 2 |
| 绒毛高度/隐窝深度 Villus height/crypt depth | 5.23 | 7.29 | 6.68 | 7.06 | 7.50 | 1.652 7 | 0.070 4 |
2.5 丁酸梭菌、葡萄糖氧化酶和迷迭香提取物组方对肉兔回肠黏膜免疫指标的影响
表6 丁酸梭菌、葡萄糖氧化酶和迷迭香提取物组方对肉兔回肠黏膜免疫指标的影响Table 6 Effects of Clostridium butyricum, glucose oxidase and rosemary extract formulas on ileum mucosal immune indices of meat rabbits |
| 项目 Items | 组别 Groups | 均方根 误差 RMSE | P值 P-value | ||||
|---|---|---|---|---|---|---|---|
| Ⅰ (对照 Control) | Ⅱ | Ⅲ | Ⅳ | Ⅴ | |||
| 分泌型免疫球蛋白A sIgA/(μg/g) | 168.80b | 219.69a | 226.34a | 250.32a | 218.45a | 42.796 0 | 0.010 7 |
| 肿瘤坏死因子-α TNF-α/(ng/g) | 3.40b | 2.95b | 3.56b | 3.38b | 4.76a | 0.960 8 | 0.008 8 |
| 干扰素-γ IFN-γ/(ng/g) | 6.70a | 4.65b | 4.82b | 4.91b | 5.29b | 1.025 0 | 0.002 1 |
| 白细胞介素-2 IL-2/(ng/mg) | 44.33 | 46.38 | 46.85 | 38.48 | 42.68 | 8.988 2 | 0.356 8 |
| 白细胞介素-6 IL-6/(ng/mg) | 778.46 | 723.85 | 736.45 | 836.02 | 783.47 | 180.658 0 | 0.749 3 |
2.6 丁酸梭菌、葡萄糖氧化酶和迷迭香提取物组方对肉兔盲肠微生物区系的影响
2.6.1 扩增子序列变体(amplicon sequence variants,ASVs)分析
2.6.2 α多样性指数分析
表7 丁酸梭菌、葡萄糖氧化酶和迷迭香提取物组方对肉兔盲肠微生物α多样性指数的影响Table 7 Effects of Clostridium butyricum, glucose oxidase and rosemary extract formulas on cecum microorganisms α diversity indexes of meat rabbits |
| 项目 Items | 组别 Groups | 均方根 误差 RMSE | P值 P-value | ||||
|---|---|---|---|---|---|---|---|
| Ⅰ (对照 Control) | Ⅱ | Ⅲ | Ⅳ | Ⅴ | |||
| ASVs数量 ASVs number | 1 038.3a | 1 058.9a | 922.1ab | 1 017.3a | 758.3b | 206.929 7 | 0.037 5 |
| Chao1指数Chao1 index | 1 041.4a | 1 063.4a | 927.1ab | 1 022.0a | 760.6b | 208.391 9 | 0.037 9 |
| Shannon指数Shannon index | 8.302 5 | 8.528 1 | 8.263 9 | 8.531 6 | 7.983 3 | 0.409 9 | 0.250 7 |
| Simpson指数 Simpson index | 0.985 4 | 0.990 8 | 0.986 6 | 0.989 9 | 0.986 0 | 0.011 6 | 0.844 4 |
| 测序深度指数 Goods_coverage index | 0.999 5 | 0.999 4 | 0.999 4 | 0.999 3 | 0.999 9 | 0.000 6 | 0.276 4 |
2.6.3 盲肠微生物物种丰富度分析
2.6.3.1 门水平分析
表8 丁酸梭菌、葡萄糖氧化酶和迷迭香提取物组方对肉兔盲肠菌群门水平的影响(前10位)Table 8 Effects of Clostridium butyricum, glucose oxidase and rosemary extract formulas on cecal microflora at phylum level of meat rabbits (top 10) |
| 项目 Items | 组别 Groups | 均方根 误差 RMSE | P值 P-value | ||||
|---|---|---|---|---|---|---|---|
| Ⅰ (对照 Control) | Ⅱ | Ⅲ | Ⅳ | Ⅴ | |||
| 厚壁菌门 Firmicutes | 24 054 | 28 619 | 29 173 | 31 397 | 31 085 | 7 751.204 0 | 0.348 0 |
| 拟杆菌门 Bacteroidota | 10 337 | 8 201 | 8 737 | 8 749 | 8 529 | 4 854.347 0 | 0.918 1 |
| 变形菌门 Proteobacteria | 3 384 | 1 080 | 776 | 1 261 | 311 | 2 138.467 0 | 0.063 4 |
| 疣微菌门 Verrucomicrobiota | 301.9c | 1 054.0bc | 1 754.9ab | 795.8bc | 2 590.5a | 1 315.486 0 | 0.013 3 |
| 酸杆菌门 Acidobacteriota | 714.1a | 373.0ab | 130.1b | 90.9b | 3.5b | 498.656 7 | 0.049 1 |
| 放线菌门 Actinobacteriota | 387.4 | 427.9 | 307.9 | 613.3 | 302.4 | 363.348 6 | 0.435 5 |
| WPS-2 | 454.8 | 141.0 | 79.3 | 80.8 | 0.0 | 320.059 0 | 0.064 6 |
| 广古菌门 Euryarchaeota | 23.8 | 172.9 | 610.3 | 403.5 | 80.3 | 438.692 8 | 0.061 7 |
| 脱硫杆菌门 Desulfobacterota | 142.6 | 236.0 | 375.8 | 209.8 | 322.5 | 268.582 7 | 0.448 7 |
| 绿弯菌门 Chloroflexi | 184.4 | 83.5 | 36.3 | 35.3 | 0.0 | 132.723 9 | 0.075 5 |
2.6.3.2 属水平分析
表9 丁酸梭菌、葡萄糖氧化酶和迷迭香提取物组方对肉兔盲肠菌群属水平的影响(前20位)Table 9 Effects of Clostridium butyricum, glucose oxidase and rosemary extract formulas on cecal microflora at genus level of meat rabbits (top 20) |
| 项目 Items | 组别 Groups | 均方根 误差 RMSE | P值 P-value | ||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Ⅰ (对照 Control) | Ⅱ | Ⅲ | Ⅳ | Ⅴ | |||||||||||
| Muribaculaceae | 8 300 | 6 476 | 6 280 | 6 763 | 7 539 | 4 995.129 0 | 0.922 8 | ||||||||
| 梭菌属_UCG-014 Clostridium_UCG-014 | 3 328.3 | 4 172.9 | 4 063.6 | 5 137.5 | 3 784.5 | 1 991.468 0 | 0.476 3 | ||||||||
| 克曼菌属 Akkermansia | 301.0c | 1 023.3bc | 1 754.9ab | 795.8bc | 2 590.5a | 1 314.927 0 | 0.012 9 | ||||||||
| 果胶单核菌属 Monoglobus | 509.6c | 753.8bc | 1 273.1bc | 1 761.3ab | 2 695.9a | 1 100.171 0 | 0.002 7 | ||||||||
| 梭菌属vadinBB60群 Clostridium_vadinBB60_group | 1 921.1 | 2 254.4 | 2 477.3 | 2 505.5 | 820.9 | 1 377.491 0 | 0.110 9 | ||||||||
| 马赛菌属Massilia | 1 269.5a | 111.1b | 66.1b | 71.9b | 0.0b | 794.456 3 | 0.012 7 | ||||||||
| V9D2013群 V9D2013_group | 968.3 | 1 370.3 | 1 265.6 | 1 774.1 | 1 445.1 | 962.882 3 | 0.573 8 | ||||||||
| 瘤胃球菌属 Ruminococcus | 1 944.1 | 1 964.1 | 1 773.1 | 2 533.8 | 1 971.0 | 809.376 9 | 0.409 6 | ||||||||
| NK4A214群 NK4A214_group | 853.0 | 1 106.9 | 1 202.0 | 1 296.1 | 1 615.5 | 689.683 7 | 0.287 9 | ||||||||
| 瘤胃球菌科Ruminococcaceae | 964.4 | 838.9 | 906.8 | 651.6 | 923.6 | 767.129 7 | 0.932 7 | ||||||||
| 优杆菌属西雷姆群 Eubacterium_siraeum_group | 1 142.9 | 1 815.5 | 1 597.6 | 1 649.4 | 1 436.8 | 571.375 0 | 0.199 7 | ||||||||
| 青枯菌属 Ralstonia | 10.5 | 3.6 | 166.9 | 326.1 | 4.4 | 425.219 0 | 0.478 4 | ||||||||
| 克里斯滕森菌科R-7群 Christensenellaceae_R-7_group | 649.4 | 692.6 | 691.8 | 760.4 | 632.0 | 475.502 2 | 0.985 8 | ||||||||
| 疣微菌属UCG-010 Verrucomicrobium_UCG-010 | 544.9b | 767.0ab | 1 024.0a | 1 030.0a | 1 011.9a | 373.533 2 | 0.048 8 | ||||||||
| WPS-2 | 454.8 | 141.0 | 79.3 | 80.8 | 0.0 | 320.059 0 | 0.064 6 | ||||||||
| 甲烷芽孢杆菌属 Methanobrevibacter | 23.8 | 168.8 | 607.8 | 398.5 | 78.8 | 434.465 3 | 0.059 3 | ||||||||
| 芽孢杆菌属 Bacillus | 443.4 | 107.9 | 192.8 | 262.4 | 0.0 | 413.886 3 | 0.287 7 | ||||||||
| 考拉杆菌属 Phascolarctobacterium | 106.8 | 354.1 | 41.0 | 121.1 | 31.9 | 288.964 7 | 0.186 6 | ||||||||
| 理研菌科RC9肠道群 Rikenellaceae_RC9_gut_group | 390.8 | 216.6 | 260.8 | 513.5 | 37.1 | 459.145 8 | 0.314 3 | ||||||||
| 鞘氨醇单胞菌属 Sphingomonas | 359.9 | 191.5 | 68.4 | 46.5 | 0.0 | 262.990 2 | 0.065 4 | ||||||||

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