1 材料与方法
1.1 试验材料
1.2 试验设计
1.3 试验饲粮
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) % |
| 项目Items | 含量Content |
|---|---|
| 原料Ingredients | |
| 玉米Corn | 69.00 |
| 豆粕Soybean meal | 21.00 |
| 石粉Limestone | 2.20 |
| 磷酸氢钙CaHPO4 | 2.80 |
| 氯化钠NaCl | 0.30 |
| 预混料Premix1) | 4.70 |
| 合计Total | 100.00 |
| 营养水平Nutrient levels2) | |
| 代谢能ME/(MJ/kg) | 11.00 |
| 粗蛋白质CP | 16.10 |
| 钙Ca | 2.60 |
| 总磷TP | 0.42 |
| 有效磷AP | 0.22 |
| 蛋氨酸Met | 0.45 |
| 赖氨酸Lys | 0.62 |
1)预混料为每千克饲粮提供 The premix provided the following per kg of the diet:VA 8 000 IU,VD3 2 800 IU,VE 23 IU,VB2 4.5 mg,VB5 2 mg,VB6 0.2 mg,VB12 0.02 mg,VK3 2.6 mg,生物素 biotin 0.007 5 mg,叶酸 folic acid 0.05 mg,泛酸 pantothenic acid 15 mg,烟酸 nicotinic acid 30.5 mg,Cu (as copper sulfate) 17.5 mg,Fe (as ferrous sulfate) 60 mg,Mn (as manganese sulfate) 65 mg,Zn (as zinc sulfate) 70 mg,I (as potassium iodide) 0.75 mg,Se (as sodium selenite) 0.2 mg。 |
1.4 饲养管理
1.5 样品采集及处理
1.6 测定指标及方法
1.6.1 生长性能
1.6.2 血清生化指标
1.6.3 肠道形态结构
1.6.4 回肠免疫和抗氧化指标
1.6.5 盲肠菌群
1.7 数据统计与分析
2 结果与分析
2.1 复合植物提取物对萧山鸡生长性能的影响
表2 复合植物提取物对萧山鸡生长性能的影响Table 2 Effects of compound plant extract on growth performance of Xiaoshan chickens |
| 项目 Items | 时间 Time | BC组 BC group | CPE200组 CPE200 group | CPE400组 CPE400 group | CPE600组 CPE600 group | P值 P-value |
|---|---|---|---|---|---|---|
体重 BW/g | 100日龄 100 days of age | 1 086.19±98.10 | 1 087.80±77.12 | 1 096.92±91.25 | 1 086.42±75.54 | 0.797 |
| 105日龄 105 days of age | 1 145.96±100.33 | 1 145.18±79.32 | 1 159.19±93.46 | 1 142.05±77.94 | 0.547 | |
| 120日龄 120 days of age | 1 330.93±119.80b | 1 347.15±102.11ab | 1 375.80±116.76a | 1 335.16±95.11b | 0.020 | |
| 135日龄 135 days of age | 1 529.43±150.81b | 1 526.52±126.30b | 1 573.50±144.29a | 1 512.33±126.64b | 0.015 | |
| 150日龄 150 days of age | 1 733.74±181.03 | 1 733.23±152.77 | 1 786.44±175.33 | 1 717.86±159.20 | 0.059 | |
| 165日龄 165 days of age | 1 863.23±214.92 | 1 833.7±190.96 | 1 906.41±207.04 | 1 809.61±184.23 | 0.268 | |
平均日采食量 ADFI/(g/d) | 105~120日龄 105 to 120 days of age | 82.61±1.62b | 84.41±0.77a | 84.43±0.92a | 82.30±1.91b | 0.004 |
| 121~135日龄 121 to 135 days of age | 100.04±2.11b | 100.28±1.41b | 102.12±0.78a | 98.84±2.03b | 0.013 | |
| 136~150日龄 136 to 150 days of age | 107.16±1.34b | 107.69±0.79ab | 108.86±0.60a | 105.82±1.75c | 0.034 | |
| 151~165日龄 151 to 165 days of age | 110.10±1.94 | 109.81±1.18 | 109.64±2.64 | 108.05±1.92 | 0.185 | |
| 105~165日龄 105 to 165 days of age | 99.98±1.70 | 100.55±0.96 | 101.26±0.93 | 98.75±1.75 | 0.074 | |
平均日增重 ADG/(g/d) | 105~120日龄 105 to 120 days of age | 12.33±3.71b | 13.46±3.43ab | 14.44±3.40a | 12.87±3.30b | <0.001 |
| 121~135日龄 121 to 135 days of age | 13.23±4.15 | 11.96±3.49 | 13.18±3.94 | 11.81±3.70 | 0.548 | |
| 136~150日龄 136 to 150 days of age | 13.62±3.80 | 13.78±4.44 | 14.20±4.61 | 13.70±4.47 | 0.801 | |
| 151~165日龄 151 to 165 days of age | 8.63±6.61 | 6.70±6.61 | 8.00±6.17 | 6.12±5.97 | 0.314 | |
| 105~165日龄 105 to 165 days of age | 11.96±2.81 | 12.45±2.74 | 11.48±2.58 | 11.13±2.45 | 0.367 | |
料重比 F/G | 105~120日龄 105 to 120 days of age | 6.76±0.67a | 6.30±0.45ab | 5.88±0.51b | 6.41±0.36ab | 0.002 |
| 121~135日龄 121 to 135 days of age | 7.72±1.07 | 8.47±0.86 | 7.83±0.76 | 8.54±1.20 | 0.239 | |
| 136~150日龄 136 to 150 days of age | 7.89±0.45 | 7.83±0.37 | 7.70±0.48 | 7.75±0.46 | 0.823 | |
| 151~165日龄 151 to 165 days of age | 13.25±3.01 | 22.29±20.00 | 13.89±1.70 | 19.44±7.58 | 0.292 | |
| 105~165日龄 105 to 165 days of age | 8.41±0.67 | 8.78±0.44 | 8.07±0.40 | 8.91±0.55 | 0.425 |
同行数据肩标相同字母或无字母表示差异不显著(P>0.05),不同字母表示差异显著(P<0.05)。下表同。 | |
In the same row, values with the same letter or no letter superscripts mean no significant difference (P>0.05), while with different letter superscripts mean significant difference (P<0.05). The same as below. |
2.2 复合植物提取物对萧山鸡血清生化指标的影响
表3 复合植物提取物对萧山鸡血清生化指标的影响Table 3 Effects of compound plant extract on serum biochemical indices of Xiaoshan chickens |
| 项目 Items | BC组 BC group | CPE200组 CPE200 group | CPE400组 CPE400 group | CPE600组 CPE600 group | P值 P-value |
|---|---|---|---|---|---|
| 总蛋白TP/(g/L) | 54.20±8.38 | 47.24±6.76 | 48.91±3.09 | 47.36±5.17 | 0.104 |
| 白蛋白ALB/(g/L) | 18.98±3.21 | 18.60±1.45 | 20.02±1.72 | 18.93±0.54 | 0.521 |
| 球蛋白GLB/(g/L) | 35.22±6.51a | 28.64±6.02b | 28.89±2.11b | 28.43±5.00b | 0.038 |
| 谷丙转氨酶ALT/(U/L) | 4.18±3.09b | 3.42±1.99b | 8.84±5.22a | 2.38±0.96b | 0.002 |
| 免疫球蛋白A IgA/(g/L) | 3.42±1.74 | 2.44±0.86 | 2.45±1.37 | 2.34±1.02 | 0.314 |
| 免疫球蛋白G IgG/(g/L) | 5.63±2.67 | 4.19±1.66 | 4.63±2.25 | 3.94±1.56 | 0.397 |
| 免疫球蛋白M IgM/(g/L) | 1.71±0.36 | 1.46±0.31 | 1.48±0.37 | 1.43±0.29 | 0.347 |
| 白细胞介素-1β IL-1β/(pg/mL) | 13.37±0.86d | 23.49±3.30b | 20.39±1.28c | 26.33±1.11a | <0.001 |
| 白细胞介素-2 IL-2/(pg/mL) | 196.71±9.43d | 292.92±31.43b | 244.98±22.12c | 321.72±30.58a | <0.001 |
| 白细胞介素-6 IL-6/(pg/mL) | 83.47±3.32d | 128.90±13.94b | 109.00±14.55c | 149.03±17.46a | <0.001 |
| 白细胞介素-10 IL-10/(pg/mL) | 18.81±0.76a | 13.38±1.35c | 14.69±0.65b | 12.32±1.03d | <0.001 |
| 干扰素-γ IFN-γ/(pg/mL) | 17.83±2.40d | 29.86±1.78b | 25.77±1.45c | 32.33±3.47a | <0.001 |
| 肿瘤坏死因子-α TNF-α/(pg/mL) | 35.52±3.42d | 48.23±3.24b | 43.36±2.52c | 54.42±3.26a | <0.001 |
2.3 复合植物提取物对萧山鸡肠道形态结构的影响
表4 复合植物提取物对萧山鸡肠道形态结构的影响Table 4 Effects of compound plant extract on intestinal morphological structure of Xiaoshan chickens |
| 项目 Items | BC组 BC group | CPE200组 CPE200 group | CPE400组 CPE400 group | CPE600组 CPE600 group | P值 P-value |
|---|---|---|---|---|---|
| 十二指肠Duodenum | |||||
| 绒毛高度Villus height/μm | 1 048.55±149.72 | 965.66±201.86 | 1 002.2±321.35 | 1 200.67±307.27 | 0.291 |
| 隐窝深度Crypt depth/μm | 166.17±57.07 | 172.31±39.65 | 150.14±47.30 | 208.03±47.13 | 0.129 |
| 绒隐比V/C | 6.74±1.72 | 5.85±1.69 | 7.79±5.09 | 6.19±2.59 | 0.622 |
| 空肠Jejunum | |||||
| 绒毛高度Villus height/μm | 897.26±144.69b | 874.30±248.79b | 1 153.29±149.93a | 867.46±181.07b | 0.012 |
| 隐窝深度Crypt depth/μm | 125.53±42.36b | 167.30±47.27ab | 159.11±49.56ab | 198.90±97.22a | 0.028 |
| 绒隐比V/C | 8.08±3.61 | 5.53±2.01 | 7.82±2.27 | 5.41±2.69 | 0.105 |
| 回肠Ileum | |||||
| 绒毛高度Villus height/μm | 832.26±141.27 | 884.01±135.95 | 763.43±129.98 | 887.61±233.64 | 0.416 |
| 隐窝深度Crypt depth/μm | 107.67±29.93 | 117.89±40.19 | 125.46±20.09 | 125.85±32.81 | 0.631 |
| 绒隐比V/C | 8.09±1.80 | 8.26±2.80 | 6.20±1.22 | 7.09±1.03 | 0.116 |
2.4 复合植物提取物对萧山鸡回肠免疫和抗氧化指标的影响
表5 复合植物提取物对萧山鸡回肠免疫指标的影响Table 5 Effects of compound plant extract on immune indices in ileum of Xiaoshan chickens |
| 项目 Items | BC组 BC group | CPE200组 CPE200 group | CPE400组 CPE400 group | CPE600组 CPE600 group | P值 P-value |
|---|---|---|---|---|---|
| 总蛋白TP/(g/L) | 7.71±6.53 | 5.19±1.17 | 5.00±1.44 | 13.64±12.17 | 0.067 |
| 白细胞介素-1β IL-1β/(pg/mL) | 1.84±0.14d | 3.08±0.25b | 2.53±0.25c | 3.47±0.61a | <0.001 |
| 白细胞介素-2 IL-2/(pg/mL) | 25.19±1.25c | 32.06±4.00b | 29.97±1.72b | 36.83±3.61a | <0.001 |
| 白细胞介素-6 IL-6/(pg/mL) | 12.41±0.41d | 20.12±2.73b | 16.61±1.49c | 22.89±1.87a | <0.001 |
| 白细胞介素-10 IL-10/(pg/mL) | 2.94±0.49a | 1.82±0.12c | 2.27±0.16b | 1.59±0.17c | <0.001 |
| 干扰素-γ IFN-γ/(pg/mL) | 4.14±0.44b | 5.68±0.55ab | 5.16±0.63b | 6.22±0.84a | <0.001 |
| 肿瘤坏死因子-α TNF-α/(pg/mL) | 3.47±0.60c | 4.16±0.38ab | 3.77±0.53b | 4.55±0.41a | <0.001 |
表6 复合植物提取物对萧山鸡回肠抗氧化指标的影响Table 6 Effects of compound plant extract on antioxidant indices in ileum of Xiaoshan chickens |
| 项目 Items | BC组 BC group | CPE200组 CPE200 group | CPE400组 CPE400 group | CPE600组 CPE600 group | P值 P-value |
|---|---|---|---|---|---|
| 总抗氧化能力T-AOC/(U/mL) | 2.82±0.21a | 2.59±0.23ab | 2.43±0.35bc | 2.12±0.24c | <0.001 |
| 丙二醛MDA/(nmol/mL) | 0.55±0.06d | 0.88±0.04b | 0.76±0.06c | 1.03±0.05a | <0.001 |
| 超氧化物歧化酶SOD/(U/mL) | 22.75±1.66a | 17.39±1.00b | 18.49±0.96b | 14.70±1.15c | <0.001 |
| 谷胱甘肽过氧化物酶GSH-Px/(U/mL) | 28.19±2.20a | 25.84±2.03b | 29.21±1.63a | 22.70±1.27c | <0.001 |
| 过氧化氢酶CAT/(U/mL) | 8.62±0.77a | 6.45±0.26c | 7.19±0.37b | 5.63±0.39d | <0.001 |
2.5 复合植物提取物对萧山鸡盲肠菌群的影响
2.5.1 盲肠宏基因组DNA测序数据质控
表7 DNA测序数据质控Table 7 Quality control of DNA sequencing data |
| 项目 Items | 样本名 Sample name | reads总数 Total number of reads/个 | 有效序列总数 Total number of effective sequences/个 | 有效序列 碱基总数 Total number of effective sequence bases/bp | 有效序列 碱基数占比 Proportion of effective sequence bases/% | GC含量 GC content/ % | Q20/ % | Q30/ % |
|---|---|---|---|---|---|---|---|---|
BC组 BC group | BC1 | 59 488 328 | 58 584 714 | 8 982 737 528 | 98.39 | 48.23 | 98.39 | 95.51 |
| BC2 | 54 794 878 | 54 071 390 | 8 274 026 578 | 98.60 | 48.12 | 98.43 | 95.50 | |
| BC3 | 66 823 296 | 65 805 488 | 10 090 317 696 | 98.38 | 49.18 | 98.37 | 95.44 | |
| BC4 | 60 073 394 | 59 231 784 | 9 071 082 494 | 98.43 | 48.56 | 98.47 | 95.69 | |
| BC5 | 57 265 984 | 56 367 980 | 8 647 163 584 | 98.31 | 49.46 | 98.34 | 95.36 | |
| BC6 | 58 290 292 | 57 366 444 | 8 801 834 092 | 98.20 | 48.34 | 98.39 | 95.54 | |
| BC7 | 63 130 978 | 62 243 372 | 9 532 777 678 | 98.31 | 47.36 | 98.47 | 95.75 | |
| BC8 | 66 249 882 | 65 446 394 | 10 003 732 182 | 98.02 | 49.59 | 98.56 | 96.00 | |
CPE400组 CPE400 group | CPE1 | 69 407 828 | 68 374 730 | 10 480 582 028 | 98.36 | 48.36 | 98.42 | 95.57 |
| CPE2 | 71 205 124 | 70 142 932 | 10 751 973 724 | 98.37 | 48.40 | 98.40 | 95.51 | |
| CPE3 | 59 368 004 | 58 514 720 | 8 964 568 604 | 98.46 | 48.58 | 98.45 | 95.64 | |
| CPE4 | 56 796 914 | 56 003 120 | 8 576 334 014 | 98.49 | 47.75 | 98.47 | 95.69 | |
| CPE5 | 76 812 922 | 75 656 234 | 11 598 751 222 | 98.35 | 48.83 | 98.37 | 95.45 | |
| CPE6 | 59 373 780 | 58 413 074 | 8 965 440 780 | 98.10 | 47.41 | 98.32 | 95.39 | |
| CPE7 | 57 531 426 | 56 716 076 | 8 687 245 326 | 98.53 | 47.86 | 98.44 | 95.59 | |
| CPE8 | 55 125 692 | 54 369 068 | 8 323 979 492 | 98.49 | 48.25 | 98.42 | 95.52 |
2.5.2 盲肠菌群多样性分析
2.5.2.1 alpha多样性分析
表8 复合植物提取物对萧山鸡盲肠菌群alpha多样性的影响Table 8 Effects of compound plant extract on alpha diversity of cecal microbiota of Xiaoshan chickens |
| 项目 Items | BC组 BC group | CPE400组 CPE400 group | P值 P-value |
|---|---|---|---|
| Chao1指数Chao1 index | 17 669.670±567.266 | 18 160.996±505.689 | 0.089 |
| ACE指数ACE index | 17 506.997±555.337 | 18 002.648±505.377 | 0.083 |
| Simpson指数Simpson index | 0.986±0.002 | 0.985±0.001 | 0.203 |
| Shannon指数Shannon index | 6.120±0.121 | 6.140±0.022 | 0.657 |
| Pielou’s_e指数Pielou’s_e index | 0.632±0.012 | 0.632±0.003 | 0.991 |
| Observer species指数Observer species index | 15 998.625±542.490 | 16 493.000±454.051 | 0.068 |
2.5.2.2 beta多样性分析
2.5.3 盲肠菌群组成分析
图2 复合植物提取物对萧山鸡盲肠菌群组成的影响A~C分别表示盲肠菌群在门、属和种水平上的OTU韦恩图,E~F分别表示盲肠菌群在门、属和种水平上的组成。 Fig.2 Effects of compound plant extract on cecal microbiota composition of Xiaoshan chickens A to C represented OTU Venn diagrams of cecal microbiota at phylum, genus and species levels, and E to F represented composition of cecal microbiota at phylum, genus and species levels, respectively. |

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