1 材料与方法
1.1 试验材料
1.2 试验设计
表1 试验饲粮组成及营养水平(干物质基础)Table 1 Composition and nutrient levels of experimental diets (DM basis) % |
| 项目 Items | 对照组 Control group | 0.2%当归组 0.2% Angelica sinensis group | |||
|---|---|---|---|---|---|
| 原料Ingredients | |||||
| 玉米Corn | 30.00 | 30.00 | |||
| 豆粕Soybean meal | 7.00 | 7.00 | |||
| 麸皮Wheat bran | 9.40 | 9.20 | |||
| 喷浆玉米皮Sprayed corn hulls | 21.00 | 21.00 | |||
| 玉米秸秆Corn straw | 30.00 | 30.00 | |||
| 当归Angelica sinensis | 0.20 | ||||
| 食盐NaCl | 0.50 | 0.50 | |||
| 石粉Limestone | 0.30 | 0.30 | |||
| 磷酸氢钙CaHPO4 | 0.30 | 0.30 | |||
| 碳酸氢钠NaHCO3 | 0.50 | 0.50 | |||
| 预混料Premix1) | 1.00 | 1.00 | |||
| 合计Total | 100.00 | 100.00 | |||
| 营养水平Nutrient levels2) | |||||
| 代谢能ME/(MJ/kg) | 9.78 | 9.78 | |||
| 粗蛋白质CP | 13.24 | 13.60 | |||
| 中性洗涤纤维NDF | 43.44 | 44.27 | |||
| 酸性洗涤纤维ADF | 16.80 | 16.29 | |||
| 粗脂肪EE | 3.65 | 3.41 | |||
| 粗灰分Ash | 9.20 | 9.79 | |||
| 钙Ca | 1.15 | 1.11 | |||
| 磷P | 0.49 | 0.53 |
1)每千克预混料含有 Each kilogram of premix contains:VA≥200 000 IU, VD3≥40 000 IU,VE≥1 000 mg,烟酰胺 nicotinamide 1 200 mg,生物素 biotin 1 200 mg,Co 11 mg,Mn 1 000 mg,Fe 1 300 mg,Zn 1 240 mg,Se 8.5 mg。 |
1.3 饲养管理
1.4 样品采集与指标测定
1.4.1 生长性能测定
1.4.2 养分表观消化率
1.4.3 瘤胃发酵参数
1.4.4 瘤胃组织形态
1.4.5 瘤胃微生物区系
1.5 数据处理与统计分析
2 结果
2.1 当归对滩羊生长性能的影响
表2 当归对滩羊生长性能的影响Table 2 Effects of Angelica sinensis on growth performance of Tan sheep |
| 项目 Items | 对照组 Control group | 0.2%当归组 0.2% Angelica sinensis group | P值 P-value |
|---|---|---|---|
| 初重IBW/kg | 32.44±3.47 | 33.01±3.27 | 0.710 |
| 末重FBW/kg | 50.29±3.77 | 52.72±4.87 | 0.229 |
| 总增重TG/kg | 17.85±1.65b | 19.71±2.06a | 0.039 |
| 平均日增重ADG/(g/d) | 198.33±18.34b | 218.94±22.87a | 0.039 |
| 平均日采食量ADFI/(g/d) | 1 776.43±0.14b | 1 778.18±2.13a | 0.018 |
| 料重比F/G | 9.03±0.88 | 8.20±0.88 | 0.050 |
同行数据肩标无字母表示差异不显著(P>0.05),不同大写字母和小写字母分别表示差异极显著(P<0.01)和显著(P<0.05)。下表同。 | |
In the same column, values with no superscript letter mean no significant difference (P>0.05), while with different uppercase and lowercase superscript letters mean extremely significant difference (P<0.01) and significant difference (P<0.05), respectively. The same as below. |
2.2 当归对滩羊养分表观消化率的影响
表3 当归对滩羊养分表观消化率的影响Table 3 Effects of Angelica sinensis on apparent digestibility of nutrients of Tan sheep % |
| 项目 Items | 对照组 Control group | 0.2%当归组 0.2% Angelica sinensis group | P值 P-value |
|---|---|---|---|
| 干物质DM | 81.69±0.66B | 85.93±0.21A | <0.001 |
| 粗蛋白质CP | 66.95±1.60b | 73.45±3.43a | 0.041 |
| 中性洗涤纤维NDF | 46.49±1.91B | 55.32±1.55A | 0.003 |
| 酸性洗涤纤维ADF | 16.11±4.84b | 33.35±6.67a | 0.022 |
| 粗脂肪EE | 81.64±3.96 | 87.28±1.49 | 0.082 |
| 粗灰分Ash | 30.70±0.06B | 41.09±0.06A | <0.001 |
2.3 当归对滩羊瘤胃发酵参数的影响
表4 当归对滩羊瘤胃发酵参数的影响Table 4 Effects of Angelica sinensis on rumen fermentation parameters of Tan sheep |
| 项目 Items | 对照组 Control group | 0.2%当归组 0.2% Angelica sinensis group | P值 P-value |
|---|---|---|---|
| pH | 6.62±0.09b | 6.77±0.09a | 0.030 |
| 氨态氮NH3-N/(mg/dL) | 19.68±0.90a | 13.54±5.40b | 0.036 |
| 乙酸Acetic acid/(mmol/L) | 20.28±2.33B | 28.75±3.77A | 0.003 |
| 丙酸Propionic acid/(mmol/L) | 10.02±0.93B | 16.59±1.19A | <0.001 |
| 乙酸/丙酸Acetic acid/propionic acid | 2.02±0.11A | 1.73±0.12B | 0.004 |
| 丁酸Butyric acid/(mmol/L) | 2.56±0.63 | 2.70±0.43 | 0.707 |
| 异丁酸Isobutyric acid/(mmol/L) | 1.18±0.39 | 1.10±0.54 | 0.787 |
| 戊酸Valeric acid/(mmol/L) | 0.54±0.08 | 0.53±0.11 | 0.913 |
| 异戊酸Isovaleric acid/(mmol/L) | 1.12±0.15 | 1.01±0.32 | 0.531 |
| 总挥发性脂肪酸TVFA/(mmol/L) | 35.71±3.94B | 50.68±5.69A | 0.001 |
2.4 当归对滩羊瘤胃组织形态的影响
表5 当归对滩羊瘤胃组织形态的影响Table 5 Effects of Angelica sinensis on rumen tissue morphology of Tan sheep |
| 项目 Items | 对照组 Control group | 0.2%当归组 0.2% Angelica sinensis group | P值 P-value |
|---|---|---|---|
| 肌层厚度Muscle layer thickness/μm | 1 308.61±129.55b | 2 322.16±402.27a | 0.014 |
| 角化层厚度Stratum corneum thickness/μm | 74.78±2.20 | 82.38±7.54 | 0.169 |
| 乳头高度Papillary height/μm | 1 811.45±373.30 | 2 385.52±314.21 | 0.111 |
| 乳头宽度Papillary width/μm | 515.71±49.58 | 513.35±88.58 | 0.970 |
| 单位长度上乳头密度 Papillary density per unit length/(个/mm) | 0.76±0.13 | 1.06±0.21 | 0.106 |
2.5 当归对滩羊瘤胃微生物区系的影响
2.5.1 操作分类单元(OTUs)聚类分析
2.5.2 Sobs指数与Shannon指数稀释曲线分析
2.5.3 Alpha多样性分析
表6 瘤胃菌群Alpha多样性指数Table 6 Alpha diversity indexes of rumen microbiota |
| 项目 Items | 对照组 Control group | 0.2%当归组 0.2% Angelica sinensis group | P值 P-value |
|---|---|---|---|
| Chao1指数Chao1 index | 2 756.29±342.80 | 2 428.42±438.98 | 0.119 |
| ACE指数ACE index | 2 884.98±370.99 | 2 532.73±476.47 | 0.123 |
| 覆盖度Good’s coverage | 0.99±0.00 | 0.99±0.01 | 0.449 |
| Shannon指数Shannon index | 7.66±0.30 | 7.37±0.42 | 0.142 |
| Simpson指数Simpson index | 0.98±0.00a | 0.97±0.01b | 0.036 |
2.5.4 Beta多样性分析
2.5.5 瘤胃菌群组成及结构分析
2.5.5.1 瘤胃菌群门水平组成分析
图5 瘤胃菌群门水平组成Bacteroidota:拟杆菌门;Bacillota:芽孢杆菌门;Pseudomonadota:假单胞菌门;Patescibacteria:帕提古菌门;Verrucomicrobiota:疣微菌门;Spirochaetota:螺旋体门;Planctomycetota:浮霉菌门;Thermodesulfobacteriota:热脱硫杆菌门;Methanobacteriota:产甲烷杆菌门;Actinomycetota:放线菌门;Other:其他菌门;Unclassified:未分类菌门。图7同 the same as Fig.7。 Fig.5 Composition of rumen microbiota at phylum level |
表7 瘤胃菌群中各菌门的相对丰度Table 7 Relative abundances of bacterial phyla in rumen microbiota % |
| 项目 Items | 对照组 Control group | 0.2%当归组 0.2% Angelica sinensis group | P值 P-value |
|---|---|---|---|
| 拟杆菌门Bacteroidota | 57.36±2.85A | 42.36±8.04B | <0.001 |
| 芽孢杆菌门Bacillota | 30.87±2.12B | 38.54±2.95A | <0.001 |
| 假单胞菌门Pseudomonadota | 6.72±2.42b | 14.09±8.18a | 0.040 |
| 帕提古菌门Patescibacteria | 1.54±0.62 | 1.38±0.55 | 0.581 |
| 疣微菌门Verrucomicrobiota | 1.06±0.47 | 0.74±0.31 | 0.134 |
| 螺旋体门Spirochaetota | 0.67±0.23 | 0.86±0.68 | 0.483 |
| 浮霉菌门Planctomycetota | 0.38±0.17 | 0.44±0.19 | 0.467 |
| 热脱硫杆菌门Thermodesulfobacteriota | 0.24±0.08 | 0.24±0.07 | 0.965 |
| 产甲烷杆菌门Methanobacteriota | 0.12±0.11 | 0.22±0.16 | 0.177 |
| 放线菌门Actinomycetota | 0.15±0.06 | 0.16±0.06 | 0.889 |
| 其他菌门Other | 0.46±0.11 | 0.56±0.31 | 0.399 |
| 未分类菌门Unclassified | 0.42±0.17 | 0.40±0.23 | 0.835 |
2.5.5.2 属水平结构分析
图6 瘤胃菌群属水平组成Xylanibacter:木聚糖杆菌属;Rikenellaceae_RC9_gut_group:理研菌科RC9肠道群;Succiniclasticum:琥珀酸杆菌属;Acinetobacter:不动杆菌属;Solibacillus:土壤芽孢杆菌属;Prevotellaceae_UCG-003:普雷沃氏菌科UCG-003;Escherichia-Shigella:埃希氏菌属-志贺氏菌属复合群;Lysinibacillus:赖氨酸芽孢杆菌属;Christensenellaceae_R-7_group:克里斯滕森菌科R-7群;Veillonellaceae_UCG-001:韦荣球菌科UCG-001;Other:其他菌属;Unclassified:未分类菌属。图7同 the same as Fig.7。 Fig.6 Composition of rumen microbiota at genus level |
表8 瘤胃菌群中各菌属的相对丰度Table 8 Relative abundances of bacterial genera in rumen microbiota % |
| 项目 Items | 对照组 Control group | 0.2%当归组 0.2% Angelica sinensis group | P值 P-value |
|---|---|---|---|
| 木聚糖杆菌属Xylanibacter | 26.57±4.50A | 12.36±4.99B | <0.001 |
| 理研菌科RC9肠道群Rikenellaceae_RC9_gut_group | 15.21±4.53a | 9.56±2.10b | 0.010 |
| 琥珀酸杆菌属Succiniclasticum | 6.17±1.38 | 7.62±2.63 | 0.194 |
| 不动杆菌属Acinetobacter | 2.15±2.31 | 8.44±7.84 | 0.060 |
| 土壤芽孢杆菌属Solibacillus | 1.39±1.72 | 5.58±5.45 | 0.071 |
| 普雷沃氏菌科UCG-003 Prevotellaceae_UCG-003 | 2.90±2.45 | 3.38±2.70 | 0.716 |
| 埃希氏菌属-志贺氏菌属复合群Escherichia-Shigella | 1.26±0.98 | 3.33±3.94 | 0.187 |
| 赖氨酸芽孢杆菌属Lysinibacillus | 2.19±2.64 | 2.37±2.40 | 0.891 |
| 克里斯滕森菌科R-7群 Christensenellaceae_R-7_group | 1.69±0.82 | 1.73±1.01 | 0.925 |
| 韦荣球菌科UCG-001 Veillonellaceae_UCG-001 | 1.35±0.70 | 1.71±1.05 | 0.444 |
| 其他菌属Other | 17.94±1.99 | 19.61±5.72 | 0.456 |
| 未分类菌属Unclassified | 21.74±3.86 | 23.13±7.05 | 0.633 |
2.5.6 瘤胃发酵参数与瘤胃菌群的相关性分析
图7 瘤胃发酵参数与瘤胃菌门(A)和菌属(B)相对丰度的相关性分析Ammonia nitrogen:氨态氮;Acetic acid:乙酸;Propionic acid:丙酸;Butyric acid:丁酸;Isobutyric acid:异丁酸;Valeric acid:戊酸;Isovaleric acid:异戊酸;Acetic acid/propionic acid:乙酸/丙酸。图中红色、蓝色分别表示正、负相关,其颜色强度与相关系数大小相对应。P<0.05时则用“*”表示,P<0.01时则用“**”表示,P≤0.001时则用“***”表示。 Fig.7 Analysis of correlations among rumen fermentation parameters and relative abundances of rumen bacterial phyla (A) and genera (B) Red and blue colors indicate positive and negative correlations, respectively, with color intensity corresponding to the correlation coefficient magnitude. When P<0.05, it is indicated by “*”; when P<0.01, it is indicated by “**”; and when P≤0.001, it is indicated by “***”. |
