1 材料与方法
1.1 伦理申明
1.2 试验材料
1.3 试验设计
1.4 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 | 11.25 | ||
| 小麦Wheat | 35.00 | ||
| 高粱Sorghum | 16.00 | ||
| 豆粕Soybean meal | 14.80 | ||
| 菜籽粕Rapeseed meal | 5.00 | ||
| 葵花仁粕Sunflower seed meal | 6.00 | ||
| 玉米蛋白粉Corn gluten meal | 3.40 | ||
| 豆油Soybean oil | 4.20 | ||
| 石粉Limestone | 1.03 | ||
| 磷酸氢钙CaHPO4 | 1.46 | ||
| 食盐NaCl | 0.30 | ||
| L-赖氨酸盐酸盐L-Lys·HCl (98.5%) | 0.38 | ||
| DL-蛋氨酸DL-Met (99%) | 0.10 | ||
| L-苏氨酸L-Thr (99%) | 0.06 | ||
| 色氨酸Trp (99%) | 0.02 | ||
| 预混料Premix1) | 1.00 | ||
| 合计Total | 100.00 | ||
| 营养水平Nutrient levels2) | |||
| 氮校正代谢能MEn/(MJ/kg) | 11.92 | ||
| 粗蛋白质CP | 20.05 | ||
| 钙Ca | 0.92 | ||
| 总磷TP | 0.65 | ||
| 非植酸磷NPP | 0.41 | ||
| 赖氨酸Lys | 1.10 | ||
| 蛋氨酸Met | 0.44 | ||
| 蛋氨酸+半胱氨酸Met+Cys | 0.79 | ||
| 苏氨酸Thr | 0.74 | ||
| 色氨酸Trp | 0.18 | ||
| 异亮氨酸Ile | 0.74 |
1)预混料为每千克饲粮提供The premix provides the following per kg of the diet:VA 12 000 IU,VD3 600 IU,VE 45 IU,VK3 2.5 mg,VB1 2.4 mg,VB2 5.0 mg,VB6 2.8 mg,VB12 0.01 mg,烟酸 nicotinic acid 42 mg,泛酸 pantothenic acid 12 mg,生物素 biotin 0.12 mg,叶酸 folic acid 1.0 mg,氯化胆碱 choline chloride 1 000 mg,Fe (as ferrous sulfate) 80 mg,Cu (as copper sulfate) 7 mg,Mn (as manganese sulfate) 60 mg,Zn (as zinc sulfate) 65 mg,I (as potassium iodide) 0.60 mg,Se (as sodium selenite) 0.15 mg。 2)粗蛋白质、钙和总磷为实测值,分别参照GB/T 6432—2018、GB/T 6436—2018和GB/T 6437—2018方法测定;氮校正代谢能、氨基酸及非植酸磷为计算值,依据《中国饲料成分及营养价值表》(2022年第33版)中各原料数据计算。CP, Ca and TP are measured values according to GB/T 6432—2018, GB/T 6436—2018 and GB/T 6437—2018, respectively. MEn, amino acids and NPP are calculated values based on the ingredient data in the Tables of Feed Composition and Nutritive Values in China (33rd edition, 2022). |
1.5 饲养管理
1.6 样品采集与制备
1.7 检测指标与方法
1.7.1 生长性能
1.7.2 血浆和空肠中抗氧化指标
1.7.3 血浆和空肠中免疫指标
1.7.4 盲肠菌群结构
1.8 统计分析
2 结果与分析
2.1 酸化剂、酶制剂和益生菌联用对1~30日龄黄羽肉鸡生长性能的影响
表2 酸化剂、酶制剂和益生菌联用对1~30日龄黄羽肉鸡生长性能的影响Table 2 Effects of combined supplementation of acidifiers, enzymes and probiotics on growth performance of yellow-feathered broilers aged 1 to 30 days |
| 项目 Items | 组别Groups | SEM | P值 P-value | ||||
|---|---|---|---|---|---|---|---|
| CON | AE | AP | EP | AEP | |||
| 初始体重IBW/g | 29.10 | 29.12 | 29.09 | 29.11 | 29.09 | 0.009 | 0.720 |
| 终末体重FBW/g | 618.94 | 625.52 | 615.67 | 631.67 | 612.19 | 5.474 | 0.083 |
| 平均日采食量ADFI/g | 38.16 | 38.26 | 38.66 | 38.71 | 38.30 | 0.383 | 0.769 |
| 平均日增重ADG/g | 19.66 | 19.82 | 19.55 | 20.09 | 19.44 | 0.249 | 0.083 |
| 料重比F/G | 1.94 | 1.95 | 1.98 | 1.93 | 1.97 | 0.023 | 0.366 |
| 死亡率Mortality/% | 0.00 | 1.67 | 1.11 | 0.56 | 0.56 | 0.577 | 0.334 |
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 mean no significant difference (P>0.05), while with no identical letter mean significant difference (P<0.05). The same as below. |
2.2 酸化剂、酶制剂和益生菌联用对30日龄黄羽肉鸡血浆和空肠中抗氧化指标的影响
表3 酸化剂、酶制剂和益生菌联用对30日龄黄羽肉鸡血浆和空肠中抗氧化指标的影响Table 3 Effects of combined supplementation of acidifiers, enzymes and probiotics on antioxidant indices in plasma and jejunum of yellow-feathered broilers aged 30 days |
| 项目 Items | 组别Groups | SEM | P值 P-value | ||||
|---|---|---|---|---|---|---|---|
| CON | AE | AP | EP | AEP | |||
| 血浆Plasma | |||||||
| 谷胱甘肽过氧化物酶GPx/(U/mL) | 5 154.4 | 5 270.0 | 4 683.0 | 4 970.6 | 5 254.5 | 189.16 | 0.185 |
| 总超氧化物歧化酶T-SOD/(U/mL) | 22.34 | 21.25 | 21.41 | 22.15 | 21.60 | 0.476 | 0.316 |
| 总抗氧化力T-AOC/(U/mL) | 9.18 | 10.30 | 8.85 | 10.69 | 11.13 | 0.836 | 0.272 |
| 丙二醛MDA/(nmol/mL) | 2.48 | 3.18 | 3.12 | 3.86 | 3.31 | 0.356 | 0.259 |
| 空肠Jejunum | |||||||
| 谷胱甘肽过氧化物酶GPx/(U/mg prot) | 51.57b | 47.58b | 63.65a | 68.33a | 64.24a | 3.652 | <0.001 |
| 总超氧化物歧化酶T-SOD/(U/mg prot) | 56.51b | 63.70ab | 73.86a | 69.38a | 74.91a | 4.189 | 0.039 |
| 总抗氧化力T-AOC/(U/mg prot) | 0.21b | 0.30a | 0.33a | 0.34a | 0.33a | 0.025 | 0.005 |
| 丙二醛MDA/(nmol/mg prot) | 2.12 | 1.83 | 1.78 | 1.74 | 1.85 | 0.119 | 0.189 |
2.3 酸化剂、酶制剂和益生菌联用对30日龄黄羽肉鸡血浆和空肠中免疫指标的影响
表4 酸化剂、酶制剂和益生菌联用对30日龄黄羽肉鸡血浆和空肠中免疫指标的影响Table 4 Effects of combined supplementation of acidifiers, enzymes and probiotics on immune indices in plasma and jejunum of yellow-feathered broilers aged 30 days |
| 项目 Items | 组别Groups | SEM | P值 P-value | ||||
|---|---|---|---|---|---|---|---|
| CON | AE | AP | EP | AEP | |||
| 血浆Plasma | |||||||
| 免疫球蛋白A IgA/(ng/mL) | 8 253.06 | 8 248.33 | 8 516.11 | 9 659.17 | 8 246.11 | 246.221 | 0.300 |
| 免疫球蛋白G IgG/(μg/mL) | 47.55b | 45.00b | 45.99b | 59.79a | 48.47b | 1.501 | 0.003 |
| 免疫球蛋白M IgM/(ng/mL) | 3 133.47 | 3 241.94 | 3 292.92 | 3 988.61 | 3 375.69 | 132.715 | 0.276 |
| 空肠Jejunum | |||||||
| 白细胞介素-1β IL-1β/(pg/mg prot) | 57.85 | 59.72 | 58.55 | 56.60 | 57.27 | 4.376 | 0.892 |
| 白细胞介素-10 IL-10/(pg/mg prot) | 30.23b | 35.57ab | 32.73b | 40.77a | 37.07ab | 2.896 | 0.026 |
| 肿瘤坏死因子-α TNF-α/(pg/mg prot) | 49.04 | 48.67 | 47.91 | 48.24 | 47.14 | 4.375 | 0.858 |
| 分泌型免疫球蛋白A sIgA/(μg/mg prot) | 428.51 | 480.67 | 474.17 | 486.21 | 498.49 | 32.828 | 0.485 |
| 免疫球蛋白G IgG/(μg/mg prot) | 7.59b | 10.13a | 10.27a | 9.64a | 8.54ab | 0.636 | 0.009 |
| 新城疫抗体滴度NDV-Ab/(pg/mg prot) | 266.53 | 300.93 | 305.23 | 329.46 | 300.38 | 18.220 | 0.089 |
2.4 酸化剂、酶制剂和益生菌联用对30日龄黄羽肉鸡盲肠菌群结构的影响
2.4.1 盲肠菌群的α-多样性分析
表5 酸化剂、酶制剂和益生菌联用对30日龄黄羽肉鸡盲肠菌群α-多样性的影响Table 5 Effects of combined supplementation of acidifiers, enzymes and probiotics on α-diversity of cecal microbiota of yellow-feathered broilers aged 30 days |
| 项目 Items | 组别Groups | SEM | P值 P-value | ||||
|---|---|---|---|---|---|---|---|
| CON | AE | AP | EP | AEP | |||
| Good’s覆盖度Good’s coverage | 0.997 | 0.995 | 0.996 | 0.995 | 0.995 | 0.001 3 | 0.574 |
| Sobs指数Sobs index | 160.0 | 186.1 | 192.2 | 283.8 | 190.0 | 39.98 | 0.091 |
| Ace指数Ace index | 217.5 | 273.0 | 256.7 | 308.9 | 273.5 | 45.70 | 0.186 |
| Chao1指数Chao1 index | 209.9 | 253.9 | 253.1 | 308.5 | 258.8 | 47.76 | 0.180 |
| Shannon指数Shannon index | 2.14 | 2.15 | 2.39 | 2.70 | 2.36 | 0.368 | 0.573 |
| Simpson指数Simpson index | 0.31 | 0.33 | 0.28 | 0.19 | 0.25 | 0.071 | 0.347 |
2.4.2 盲肠菌群的β-多样性分析
2.4.3 盲肠菌群门水平和属水平的物种组成分析
图2 盲肠菌群在门水平(A)和属水平(B)上的物种组成与相对丰度Bacteroidota:拟杆菌门;Bacillota:厚壁菌门;Campylobacterota:弯曲杆菌门;Pseudomonadota:假单胞菌门;Thermodesulfobacteriota:热脱硫杆菌门;Lentisphaerota:黏胶球形菌门;Actinomycetota:放线菌门;Campylobacter:弯曲杆菌属;Escherichia:埃希氏菌属;Enterococcus:肠球菌属;Bacteroides:拟杆菌属;Phocaeicola:居海事城球杆菌属;Barnesiella:巴恩斯氏菌属;Alistipes:另枝菌属;Pseudoflavonifractor:假黄铜单胞菌属;Gemmiger:芽殖菌属;Faecalibacterium:粪杆菌属;Clostridium:梭菌属;Helicobacter:螺杆菌属;Anaerotignum:厌氧菌属;Butyricicoccus:丁酸球菌属;Blautia:布劳特氏菌属;Streptococcus:链球菌属;Phascolarctobacterium:考拉杆菌属;Others:其他。 Fig.2 Species composition and relative abundance of cecal microbiota at phylum level (A) and genus level (B) |
2.4.4 差异物种分析
图3 盲肠菌群差异物种Kruskal-Wallis秩和检验Fig.3 Kruskal-Wallis rank-sum test of different species in cecal microbiota *:P<0.05;**:P<0.01;***:P<0.001。 |
图4 盲肠菌群差异物种LEfSe分析(LDA得分>2.0)A:LEfSe分析进化分枝图 LEfSe analysis evolution cladogram;B:LDA得分分布柱状图LDA score distribution histogram。 图中各微生物名称前的字母代表其对应的分类学层级:p、c、o、f、g、s分别表示门、纲、目、科、属和种。限于篇幅,仅对各组中的关键差异物种的拉丁文进行中文注释。The letters preceding each microbial name in the figure represent their corresponding taxonomic levels: p, c, o, f, g and s denote phylum, class, order, family, genus and species, respectively. Due to space limitations, only the Latin names of key differential species in each group are annotated in Chinese. Verrucomicrobiota:疣微菌门;Streptococcus:链球菌属;Xanthomonas:黄单胞菌属;Staphylococcus_epidermidis:表皮葡萄球菌;Fusobacteriota:梭杆菌门;Fusobacterium:梭杆菌属;Weissella:魏斯氏菌属;Barnesiella:巴恩斯氏菌属;Barnesiellaceae:巴恩斯氏菌科;Morganellaceae:摩根菌科;Hyphomicrobiaceae:生丝微菌科。 Fig.4 LEfSe analysis of different species in cecal microbiota (LDA score>2.0) |

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