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 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 | 组别 Groups | ||
|---|---|---|---|
| CON | LP | SRU | |
| 原料 Ingredients | |||
| 玉米 Corn | 39.96 | 40.98 | 43.20 |
| 麦麸 Wheat bran | 3.60 | 13.20 | 2.88 |
| 豆粕 Soybean meal | 3.00 | 0.60 | |
| 棉籽粕 Cottonseed meal | 3.84 | 0.84 | 3.66 |
| 菜籽粕 Rapeseed meal | 6.00 | 0.60 | 6.00 |
| 磷酸氢钙 CaHPO4 | 0.18 | 0.06 | |
| 缓释尿素 Slow-release urea | 0.60 | ||
| 大豆油 Soybean oil | 0.60 | 0.60 | 0.60 |
| 预混料 Premix1) | 3.00 | 3.00 | 3.00 |
| 皇竹草青贮Pennisetum sinese Roxb silage | 30.00 | 30.00 | 30.00 |
| 干稻草 Straw hay | 10.00 | 10.00 | 10.00 |
| 合计 Total | 100.00 | 100.00 | 100.00 |
| 营养水平 Nutrient levels2) | |||
| 综合净能 NEmf/(MJ/kg) | 5.98 | 5.97 | 5.97 |
| 粗蛋白质 CP | 12.54 | 9.52 | 12.51 |
| 粗脂肪 EE | 3.33 | 3.59 | 3.40 |
| 粗纤维 CF | 20.84 | 23.40 | 23.14 |
| 粗灰分 Ash | 4.77 | 4.51 | 4.57 |
| 中性洗涤纤维 NDF | 32.85 | 34.73 | 32.59 |
| 酸性洗涤纤维 ADF | 19.76 | 19.21 | 19.39 |
| 钙 Ca | 0.65 | 0.64 | 0.65 |
| 磷 P | 0.52 | 0.53 | 0.51 |
1)预混料为每千克饲粮提供The premix provided the following per kg of diets:VA 3 600 IU,VD 360 IU,VE 10 IU,Cu (as copper sulfate) 6 mg,Fe (as ferrous sulfate) 48 mg,Mn (as manganese sulfate) 36 mg,Zn (as zinc sulfate) 48 mg,I (as potassium iodide) 0.36 mg,Se (as sodium selenite) 0.12 mg,Co (as cobalt chloride) 0.2 mg。 | |
2)综合净能为计算值,其他营养水平为实测值。NEmf was a calculated value, while the other nutrient levels were measured values. |
1.4 饲养管理
1.5 测定指标及方法
1.5.1 饲粮营养水平
1.5.2 生长性能
1.5.3 血清生化指标
1.5.4 瘤胃发酵参数及微生物区系
1.6 数据分析
2 结果与分析
2.1 低蛋白质饲粮中添加缓释尿素对肉牛生长性能的影响
表2 低蛋白质饲粮中添加缓释尿素对肉牛生长性能的影响Table 2 Effects of supplementing slow-release urea in a low-protein diet on growth performance of beef cattle |
| 项目 Items | 组别 Groups | 均值标准误 SEM | P值 P-value | ||
|---|---|---|---|---|---|
| CON | LP | SRU | |||
| 初重 IBW/kg | 524.17 | 520.92 | 520.5 | 5.388 | 0.960 |
| 末重 FBW/kg | 574.58 | 569.83 | 572.08 | 6.011 | 0.955 |
| 平均日增重 ADG/(g/d) | 1 440.48 | 1 350.00 | 1 473.81 | 43.965 | 0.798 |
| 干物质采食量 DMI/(kg/d) | 12.12 | 12.05 | 12.44 | 0.109 | 0.381 |
| 料重比 F/G | 8.48 | 8.88 | 8.52 | 0.263 | 0.806 |
同行数据肩标无字母或相同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 no letter or the same 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 低蛋白质饲粮中添加缓释尿素对肉牛血清生化指标的影响
表3 低蛋白质饲粮中添加缓释尿素对肉牛血清生化指标的影响Table 3 Effects of supplementing slow-release urea in a low-protein diet on serum biochemical indexes of beef cattle |
| 项目 Items | 组别 Groups | 均值标准误 SEM | P值 P-value | ||
|---|---|---|---|---|---|
| CON | LP | SRU | |||
| 谷丙转氨酶 ALT/(U/L) | 32.50 | 38.67 | 34.33 | 2.232 | 0.168 |
| 谷草转氨酶 AST/(U/L) | 96.67 | 107.67 | 90.33 | 5.414 | 0.105 |
| 谷氨酰氨基转移酶 GGT/(U/L) | 21.50 | 23.50 | 23.17 | 1.673 | 0.671 |
| 碱性磷酸酶 ALP/(U/L) | 209.17 | 267.83 | 221.17 | 28.275 | 0.328 |
| 总蛋白 TP/(g/L) | 72.37 | 69.68 | 72.40 | 1.159 | 0.198 |
| 白蛋白 ALB/(g/L) | 35.73 | 35.82 | 35.67 | 0.985 | 0.994 |
| 尿素氮 UN/(mmol/L) | 4.72a | 3.66b | 4.64a | 0.147 | <0.001 |
| 高密度脂蛋白胆固醇 HDL-C/(mmol/L) | 2.01 | 2.30 | 2.08 | 0.105 | 0.158 |
| 低密度脂蛋白胆固醇 LDL-C/(mmol/L) | 0.75 | 0.84 | 0.68 | 0.089 | 0.471 |
| 葡萄糖 GLU/(mmol/L) | 4.06 | 4.08 | 4.12 | 0.151 | 0.965 |
2.3 低蛋白质饲粮中添加缓释尿素对肉牛瘤胃发酵参数的影响
表4 低蛋白质饲粮中添加缓释尿素对肉牛瘤胃发酵参数的影响Table 4 Effects of supplementing slow-release urea in a low-protein diet on ruminal fermentation parameters of beef cattle |
| 项目 Items | 组别 Groups | 均值标准误 SEM | P值 P-value | ||
|---|---|---|---|---|---|
| CON | LP | SRU | |||
| pH | 6.71 | 6.69 | 6.72 | 0.023 | 0.788 |
| 氨态氮 NH3-N/(μg/mL) | 26.37b | 23.33b | 33.95a | 1.435 | <0.001 |
| 微生物蛋白 MCP/(mg/mL) | 4.95a | 3.57b | 4.99a | 0.246 | 0.001 |
| 总挥发性脂肪酸 TVFA/(mmol/L) | 68.22 | 63.03 | 73.67 | 3.115 | 0.085 |
| 乙酸 Acetate/% | 70.63 | 70.62 | 69.68 | 0.372 | 0.150 |
| 丙酸 Propionate/% | 21.80 | 22.37 | 22.37 | 0.392 | 0.510 |
| 异丁酸 Isobutyrate/% | 1.33a | 1.17b | 1.29a | 0.038 | 0.028 |
| 丁酸 Butyrate/% | 4.56ab | 4.34b | 4.88a | 0.115 | 0.016 |
| 异戊酸 Isovalerate/% | 1.17 | 1.07 | 1.29 | 0.065 | 0.087 |
| 戊酸 Valerate/% | 0.51 | 0.43 | 0.50 | 0.035 | 0.192 |
| 乙丙比 Acetate/propionate | 3.24 | 3.16 | 3.13 | 0.069 | 0.490 |
2.4 低蛋白质饲粮中添加缓释尿素对肉牛瘤胃微生物区系的影响
2.4.1 Alpha多样性分析
表5 低蛋白质饲粮中添加缓释尿素对肉牛瘤胃微生物Alpha多样性的影响Table 5 Effects of supplementing slow-release urea in a low-protein diet on Alpha diversity of rumen microbiota of beef cattle |
| 项目 Items | 组别 Groups | 均值标准误 SEM | P值 P-value | ||
|---|---|---|---|---|---|
| CON | LP | SRU | |||
| ASVs数量 ASVs number | 1 836.67 | 1 756.50 | 1 819.17 | 96.921 | 0.830 |
| ACE指数 ACE index | 1 875.41 | 1 785.59 | 1 847.76 | 106.613 | 0.832 |
| Chao指数 Chao index | 1 855.64 | 1 771.88 | 1 830.47 | 101.489 | 0.838 |
| Shannon指数 Shannon index | 6.75 | 6.73 | 6.77 | 0.062 | 0.915 |
| Simpson指数 Simpson index | 0.002 | 0.003 | 0.002 | 0.000 1 | 0.756 |
| Good’s Coverage指数 Good’s Coverage index | 0.998 | 0.998 | 0.998 | 0.001 2 | 0.763 |
2.4.2 Venn图分析
2.4.3 PCoA图分析
2.4.4 瘤胃微生物组成分析
表6 低蛋白质饲粮中添加缓释尿素对肉牛瘤胃微生物组成的影响(门水平相对丰度)Table 6 Effects of supplementing slow-release urea in a low-protein diet on rumen microbial composition of beef cattle (relative abundance at phylum level) |
| 项目 Items | 组别 Groups | 均值标准误 SEM | P值 P-value | ||
|---|---|---|---|---|---|
| CON | LP | SRU | |||
| 芽孢杆菌门 Bacillota | 48.94 | 48.45 | 47.08 | 2.598 | 0.871 |
| 拟杆菌门 Bacteroidota | 44.28 | 43.67 | 46.38 | 2.446 | 0.718 |
| 髌骨菌门 Patescibacteria | 1.48b | 2.58a | 1.98b | 0.244 | 0.020 |
| 螺旋菌门 Spirochaetota | 1.71 | 1.43 | 1.57 | 0.285 | 0.779 |
| 纤维杆菌门 Fibrobacterota | 0.91 | 0.68 | 0.63 | 0.162 | 0.446 |
| 疣微菌门 Verrucomicrobiota | 0.61 | 0.78 | 0.65 | 0.099 | 0.465 |
| 假单胞菌门 Pseudomonadota | 0.45 | 1.03 | 0.35 | 0.370 | 0.393 |
| 放线菌门 Actinomycetota | 0.60 | 0.59 | 0.44 | 0.134 | 0.643 |
| 热脱硫杆菌门 Thermodesulfobacteriota | 0.42 | 0.34 | 0.41 | 0.054 | 0.541 |
| unclassified_k__norank_d__Bacteria | 0.42 | 0.23 | 0.32 | 0.084 | 0.333 |
| 其他 Others | 0.18 | 0.22 | 0.19 | 0.048 | 0.814 |
表7 低蛋白质饲粮中添加缓释尿素对肉牛瘤胃微生物组成的影响(属水平相对丰度)Table 7 Effects of supplementing slow-release urea in a low-protein diet on rumen microbial composition of beef cattle (relative abundance at genus level) |
| 项目 Items | 组别 Groups | 均值标准误 SEM | P值 P-value | ||
|---|---|---|---|---|---|
| CON | LP | SRU | |||
| 木聚糖杆菌属 Xylanibacter | 11.67 | 13.29 | 14.99 | 2.165 | 0.568 |
| 理研菌科RC9肠道群 Rikenellaceae_RC9_gut_group | 11.05 | 10.75 | 9.52 | 0.747 | 0.337 |
| norank_f__F082 | 7.07 | 6.29 | 6.23 | 0.665 | 0.616 |
| 瘤胃球菌属 Ruminococcus | 3.32 | 4.07 | 2.93 | 0.593 | 0.405 |
| NK4A214群 NK4A214_group | 3.92a | 2.77b | 3.03ab | 0.300 | 0.039 |
| 克里斯滕森菌科R7群 Christensenellaceae_R-7_group | 3.58 | 2.72 | 3.13 | 0.376 | 0.300 |
| 拟杆菌目RF16群 Bacteroidales_RF16_group | 3.00 | 3.07 | 2.46 | 0.336 | 0.393 |
| 普雷沃氏菌科UCG-003 Prevotellaceae_UCG-003 | 2.18 | 1.94 | 3.13 | 0.544 | 0.294 |
| norank_o__RF39 | 2.01 | 2.67 | 2.36 | 0.250 | 0.206 |
| 未分类的梭菌纲 unclassified_c__Clostridia | 2.38 | 2.48 | 2.15 | 0.262 | 0.668 |
| 梭菌纲UCG-014 Clostridia_UCG-014 | 1.65 | 2.29 | 2.72 | 0.436 | 0.251 |
| Saccharofermentans | 2.15 | 2.10 | 1.90 | 0.258 | 0.775 |
| 产粪甾醇真杆菌群 [Eubacterium]_coprostanoligenes_group | 1.73 | 2.29 | 1.99 | 0.207 | 0.199 |
| 毛螺菌科NK3A20群 Lachnospiraceae_NK3A20_group | 2.56a | 1.37b | 1.74b | 0.212 | 0.004 |
| norank_f__UCG-010 | 2.17 | 1.69 | 1.63 | 0.185 | 0.109 |
| 其他 Others | 39.57 | 40.20 | 40.10 | 1.111 | 0.912 |

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