1 材料与方法
1.1 SBPE和氧化大豆油的制备
1.2 SBPE活性成分的定性鉴定
1.2.1 SBPE前处理
1.2.2 超高效液相色谱-串联质谱(UPLC-MS/MS)
1.2.3 数据处理和化合物鉴定
1.3 试验动物、分组及饲粮
表1 基础饲粮组成及营养水平(风干基础)Table 1 Composition and nutrient levels of basal diets (air-dry basis) % |
| 项目 Items | 1~21日龄 1 to 21 days of age | 22~42日龄 22 to 42 days of age | 43~63日龄 43 to 63 days of age |
|---|---|---|---|
| 原料Ingredients | |||
| 玉米Corn | 53.88 | 59.81 | 69.40 |
| 麸皮Wheat bran | 2.20 | 2.47 | 1.83 |
| 豆粕Soybean meal | 22.53 | 22.28 | 15.66 |
| 全脂大豆Full-fat soybean | 6.00 | 3.80 | 2.97 |
| 大豆油Soybean oil | 2.00 | 2.50 | 3.00 |
| 棉籽粕Cottonseed meal | 9.00 | 5.10 | 3.37 |
| 磷酸氢钙CaHPO4 | 2.19 | 2.00 | 1.70 |
| 石粉Limestone | 1.23 | 1.10 | 1.05 |
| 氯化钠NaCl | 0.35 | 0.35 | 0.35 |
| 预混料Premix1) | 0.50 | 0.50 | 0.50 |
| DL-蛋氨酸DL-Met | 0.12 | 0.09 | 0.07 |
| L-赖氨酸L-Lys | 0.10 | ||
| 合计Total | 100.00 | 100.00 | 100.00 |
| 营养水平Nutrient levels2) | |||
| 代谢能ME/(MJ/kg) | 12.12 | 12.54 | 12.96 |
| 粗蛋白质CP | 20.45 | 18.46 | 16.03 |
| 钙Ca | 1.29 | 1.40 | 1.22 |
| 总磷TP | 0.81 | 0.82 | 0.72 |
| DL-蛋氨酸DL-Met | 0.46 | 0.38 | 0.34 |
| 蛋氨酸+胱氨酸Met+Cys | 0.85 | 0.72 | 0.65 |
| L-赖氨酸L-Lys | 1.05 | 0.98 | 0.85 |
1)预混料为每千克饲粮提供 The premix provided the following per kg of diets:VD3 1 250 IU,VA 7 500 IU,VK3 0.6 mg,DL-α-生育酚乙酸酯 DL-α-tocopherol acetate 30 mg,盐酸硫胺 thiamine hydrochloride 3 mg,核黄素 riboflavin 6 mg,叶酸 folic acid 0.75 mg,烟酸 niacin 35 mg,Zn 75 mg,Mn 100 mg,Fe 400 mg,Cu 2.25 mg。 | |
2)粗蛋白质(GB/T 6432—2018)、钙(GB/T 6436—2018)和总磷(GB/T 6437—2018)为实测值,其余为计算值[参考中国饲料成分及营养价值表(2019年第30版)]。CP (GB/T 6432—2018), Ca (GB/T 6436—2018) and TP (GB/T 6437—2018) were measured values, while the others were calculated values [referred to Tables of Feed Composition and Nutritive in China (30th edition, 2019)]. |
1.4 饲养管理
1.5 样品采集及指标测定
1.5.1 屠宰性能
1.5.2 肉品质
1.5.3 肝脏抗氧化指标
1.5.4 肠道黏膜形态结构
1.5.5 盲肠内容物VFA含量
1.5.6 回肠微生物16S rDNA扩增子测序
1.6 数据统计与分析
2 结果与分析
2.1 SBPE活性成分的定性鉴定
表2 二级质谱筛选中相对含量较大的物质Table 2 Substances with relatively large content in secondary mass spectrometry screening |
| 化合物名称 Compound name | 保留时间 RT/min | 质荷比 m/z | 加合物 Adducts | 相对含量 Relative content/% | 分类 Classification | 分子式 Formula |
|---|---|---|---|---|---|---|
| 3-(3,5-dimethyl-7-oxo-7H-furo[3,2-g] chromen-6-yl)-N-(2-fluorophenyl)propanamide | 0.92 | 380.14 | [M+H]+ | 4.47 | 含氟芳香族 | C22H18FNO4 |
| 7-methoxy-6-(1,2,3-trihydroxy-3- methylbutyl)chromen-2-one | 7.42 | 317.10 | [M+Na]+ | 2.36 | 香豆素 | C15H18O6 |
| 5,7-dihydroxy-2-(3-hydroxy-4- methoxyphenyl)-2,3-dihydrochromen-4-one | 8.14 | 303.08 | [M+H]+ | 8.03 | 黄酮类 | C16H14O6 |
| 橙皮素Hesperetin | 12.08 | 303.08 | [M+H]+ | 3.65 | 黄酮类 | C16H14O6 |
| 倒捻子素Mangostin | 5.74 | 433.16 | [M+Na]+ | 2.02 | 黄酮类 | C24H26O6 |
| 枸橘苷Poncirin | 11.99 | 617.19 | [M+Na]+ | 1.91 | 黄酮类 | C28H34O14 |
| 苏木精Hematoxylin | 6.63 | 303.08 | [M+H]+ | 1.88 | 黄酮类 | C16H14O6 |
| 樱花素Sakuranetin | 9.07 | 287.08 | [M+H]+ | 1.49 | 黄酮类 | C16H14O5 |
| 辛夷脂素Fargesin | 20.64 | 371.15 | [M+H]+ | 6.10 | 木脂素 | C21H22O6 |
| 五味子酚Schisanhenol | 5.40 | 425.19 | [M+Na]+ | 2.44 | 木脂素 | C23H30O6 |
| 亮肽素Leupeptin | 14.20 | 427.31 | [M+H]+ | 2.98 | 肽醛类 | C20H38N6O4 1/2(H2SO4) |
| Tonantzitlolone A | 8.56 | 487.26 | [M+Na]+ | 2.24 | 萜类 | C26H40O7 |
| 盐酸吐根碱Emetine hydrochloride | 12.50 | 539.27 | [M+Na]+ | 1.59 | 生物碱类 | C29H41CIN2O4 |
| 血根碱Sanguinarine | 0.98 | 333.11 | [M+H]+ | 1.33 | 生物碱类 | C20H14NO4 |
| 胸腺嘧啶Thymine | 3.49 | 127.05 | [M+H]+ | 1.31 | 核苷类 | C5H6N2O2 |
| 柠檬酸Citric acid | 1.56 | 191.01 | [M-H]- | 8.43 | 三羧酸 | C6H8O7 |
| 儿茶酚Catechol | 3.55 | 109.02 | [M-H]- | 10.44 | 多酚 | C6H6O2 |
2.2 SBPE对黄羽肉鸡屠宰性能的影响
表3 SBPE对黄羽肉鸡屠宰性能的影响Table 3 Effects of SBPE on slaughter performance of yellow-feathered broilers |
| 项目 Items | BC组 BC group | OX组 OX group | SBPE添加量 Addition amount of SBPE/% | SEM | P值 P-value | ||
|---|---|---|---|---|---|---|---|
| 0.1 | 0.2 | 0.4 | |||||
| 活重Live weight/g | 1 966.23 | 1 876.08 | 1 908.88 | 1 982.50 | 1 918.70 | 29.92 | 0.807 |
| 屠宰率Dressed percentage/% | 92.50 | 93.13 | 93.08 | 94.03 | 93.33 | 0.30 | 0.621 |
| 半净膛率 Percentage of half-eviscerated yield/% | 83.84 | 84.66 | 84.17 | 85.03 | 83.15 | 0.27 | 0.210 |
| 全净膛率Percentage of eviscerated yield/% | 67.52 | 68.71 | 67.58 | 68.75 | 68.18 | 0.26 | 0.393 |
| 胸肌率Percentage of breast muscle/% | 15.82 | 15.11 | 15.86 | 15.82 | 15.94 | 0.22 | 0.756 |
| 腿肌率Percentage of leg muscle/% | 21.61 | 21.64 | 21.34 | 21.74 | 21.53 | 0.14 | 0.934 |
| 腹脂率Percentage of abdominal fat/% | 3.94 | 4.00 | 3.84 | 4.27 | 3.88 | 0.13 | 0.872 |
同行数据肩标不同小写字母表示差异显著(P<0.05),不同大写字母表示差异极显著(P<0.01),相同小写字母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), and with different capital letter superscripts mean extremely significant difference (P<0.01), while with the same small letter or no letter superscripts mean no significant difference (P>0.05). The same as below. |
2.3 SBPE对黄羽肉鸡肉品质的影响
表4 SBPE对黄羽肉鸡肉品质的影响Table 4 Effects of SBPE on meat quality of yellow-feathered broilers |
| 项目 Items | BC组 BC group | OX组 OX group | SBPE添加量 Addition amount of SBPE/% | SEM | P值 P-value | ||
|---|---|---|---|---|---|---|---|
| 0.1 | 0.2 | 0.4 | |||||
| 剪切力Shear force/N | 55.04ab | 43.38b | 53.67ab | 63.85a | 53.24ab | 2.03 | 0.034 |
| pH1 h | 5.73 | 5.85 | 5.78 | 5.61 | 5.79 | 0.04 | 0.305 |
| pH24 h | 5.41 | 5.38 | 5.32 | 5.34 | 5.35 | 0.02 | 0.807 |
| 1 h肉色Meat color at 1 h | 71.45 | 74.16 | 76.36 | 73.69 | 72.28 | 0.63 | 0.123 |
| 24 h肉色Meat color at 24 h | 61.20 | 60.90 | 61.36 | 61.41 | 62.33 | 0.49 | 0.927 |
| 滴水损失率Drip loss/% | 5.61 | 6.90 | 7.23 | 6.58 | 6.74 | 0.24 | 0.333 |
2.4 SBPE对黄羽肉鸡肝脏抗氧化指标的影响
表5 SBPE对黄羽肉鸡肝脏抗氧化指标的影响Table 5 Effects of SBPE on antioxidant indices in liver of yellow-feathered broilers |
| 项目 Items | BC组 BC group | OX组 OX group | SBPE添加量 Addition amount of SBPE/% | SEM | P值 P-value | ||
|---|---|---|---|---|---|---|---|
| 0.1 | 0.2 | 0.4 | |||||
| 丙二醛MDA/(nmol/mg prot) | 0.42 | 0.51 | 0.49 | 0.49 | 0.47 | 0.02 | 0.559 |
| 总抗氧化能力T-AOC/(μmol/g prot) | 35.53 | 35.58 | 49.82 | 39.17 | 36.41 | 1.97 | 0.123 |
| 过氧化氢酶CAT/(U/mg prot) | 12.46 | 13.47 | 11.90 | 12.42 | 11.70 | 0.43 | 0.744 |
| 超氧化物歧化酶SOD/(U/mg prot) | 41.67 | 46.61 | 42.54 | 38.40 | 41.83 | 1.98 | 0.800 |
| 谷胱甘肽过氧化物酶 GSH-Px/(U/mg prot) | 4.01 | 4.92 | 4.56 | 4.17 | 4.76 | 0.18 | 0.428 |
2.5 SBPE对黄羽肉鸡肠道黏膜形态结构的影响
表6 SBPE对黄羽肉鸡肠道黏膜形态结构的影响Table 6 Effects of SBPE on intestinal mucosal morphology and structure of yellow-feathered broilers |
| 项目 Items | BC组 BC group | OX组 OX group | SBPE添加量 Addition amount of SBPE/% | SEM | P值 P-value | |||
|---|---|---|---|---|---|---|---|---|
| 0.1 | 0.2 | 0.4 | ||||||
十二指肠 Duodenum | 绒毛高度Villus height/μm | 1 767.42 | 1 786.19 | 1 838.12 | 1 828.93 | 1 954.36 | 27.29 | 0.223 |
| 隐窝深度Crypt depth/μm | 261.29 | 219.15 | 229.29 | 250.41 | 240.39 | 5.88 | 0.176 | |
| 绒毛高度/隐窝深度V/C | 6.90 | 8.13 | 8.11 | 7.61 | 8.32 | 0.19 | 0.118 | |
空肠 Jejunum | 绒毛高度Villus height/μm | 1 324.46b | 1 303.82b | 1 372.75ab | 1 468.08a | 1 396.31ab | 18.92 | 0.042 |
| 隐窝深度Crypt depth/μm | 223.01 | 212.77 | 201.68 | 231.45 | 201.46 | 4.99 | 0.234 | |
| 绒毛高度/隐窝深度V/C | 6.00 | 6.28 | 6.83 | 6.48 | 6.86 | 0.14 | 0.283 | |
回肠 Ileum | 绒毛高度Villus height/μm | 887.08 | 905.33 | 934.85 | 969.56 | 911.80 | 15.67 | 0.527 |
| 隐窝深度Crypt depth/μm | 167.68 | 158.92 | 170.77 | 173.60 | 178.50 | 3.09 | 0.375 | |
| 绒毛高度/隐窝深度V/C | 5.36 | 5.82 | 5.53 | 5.60 | 5.16 | 0.11 | 0.422 | |
2.6 SBPE对黄羽肉鸡盲肠内容物VFA含量的影响
表7 SBPE对黄羽肉鸡盲肠内容物VFA含量的影响Table 7 Effects of SBPE on VFA content in cecal content of yellow-feathered broilersmg/g |
| 项目 Items | BC组 BC group | OX组 OX group | SBPE添加量 Addition amount of SBPE/% | SEM | P值 P-value | ||
|---|---|---|---|---|---|---|---|
| 0.1 | 0.2 | 0.4 | |||||
| 乙酸Acetate | 13.04 | 12.23 | 12.21 | 11.54 | 14.09 | 0.53 | 0.612 |
| 丙酸Propionate | 4.35 | 3.92 | 3.51 | 3.82 | 4.73 | 0.16 | 0.134 |
| 异丁酸Isobutyrate | 0.33 | 0.27 | 0.31 | 0.29 | 0.32 | 0.01 | 0.634 |
| 丁酸Butyrate | 4.14 | 3.77 | 3.78 | 3.86 | 5.39 | 0.27 | 0.248 |
| 异戊酸Isovalerate | 0.49 | 0.39 | 0.46 | 0.42 | 0.46 | 0.02 | 0.508 |
| 戊酸Valerate | 0.31 | 0.28 | 0.40 | 0.33 | 0.32 | 0.02 | 0.260 |
2.7 SBPE对黄羽肉鸡回肠微生物的影响
2.7.1 SBPE对黄羽肉鸡回肠微生物α多样性的影响
表8 SBPE对黄羽肉鸡回肠微生物α多样性的影响Table 8 Effects of SBPE on microbial α diversity in ileum of yellow-feather broilers |
| 项目 Items | BC组 BC group | OX组 OX group | SBPE添加量 Addition amount of SBPE/% | SEM | P值 P-value | ||
|---|---|---|---|---|---|---|---|
| 0.1 | 0.2 | 0.4 | |||||
| Chao1指数Chao1 index | 579.55ABa | 493.28BCb | 637.44Aa | 462.53Cb | 461.86Cb | 15.74 | <0.001 |
| 操作分类单元OTUs | 574.50ABa | 490.30BCb | 631.70Aa | 458.80Cb | 458.70Cb | 15.56 | <0.001 |
| 香农指数Shannon index | 4.94 | 5.12 | 5.00 | 4.53 | 4.83 | 0.09 | 0.252 |
| 辛普森指数Simpson index | 0.87 | 0.91 | 0.89 | 0.86 | 0.88 | 0.01 | 0.410 |
2.7.2 黄羽肉鸡回肠微生物OTUs分析
2.7.3 SBPE对黄羽肉鸡回肠微生物组成的影响
图2 SBPE对黄羽肉鸡回肠微生物组成的影响a:门水平 at phylum level;b:属水平 at genus level。 Fig.2 Effects of SBPE on ileal microbiota composition of yellow-feathered broilers |
表9 SBPE对黄羽肉鸡回肠微生物在门水平上相对丰度的影响(平均相对丰度>5%)Table 9 Effects of SBPE on relative abundance of ileal microbiota at phylum level of yellow-feather broilers (average relative abundance>5%) % |
| 项目 Items | BC组 BC group | OX组 OX group | SBPE添加量 Addition amount of SBPE/% | SEM | P值 P-value | ||
|---|---|---|---|---|---|---|---|
| 0.1 | 0.2 | 0.4 | |||||
| 厚壁菌门Firmicutes | 60.98b | 68.97ab | 69.94ab | 74.93a | 75.28a | 1.60 | 0.025 |
| 变形菌门Proteobacteria | 25.70a | 21.29ab | 15.83b | 14.11b | 13.25b | 1.36 | 0.011 |
| 蓝藻门Cyanobacteria | 6.22 | 1.76 | 4.35 | 3.10 | 3.12 | 0.63 | 0.224 |
| 拟杆菌门Bacteroidota | 5.14 | 6.81 | 7.60 | 6.87 | 7.70 | 0.45 | 0.399 |
表10 SBPE对黄羽肉鸡回肠微生物在属水平上相对丰度(>1%)的影响Table 10 Effects of SBPE on relative abundance of ileal microbiota at genus level of yellow-feather broilers (average relative abundance>1%) % |
| 项目 Items | BC组 BC group | OX组 OX group | SBPE添加量 Addition amount of SBPE/% | SEM | P值 P-value | ||
|---|---|---|---|---|---|---|---|
| 0.1 | 0.2 | 0.4 | |||||
| 乳杆菌属Lactobacillus | 27.79Bc | 37.97ABbc | 36.25ABc | 50.83Aa | 48.78Aab | 2.13 | 0.001 |
| 大肠杆菌-志贺菌属Escherichia-Shigella | 9.83 | 6.50 | 9.75 | 9.80 | 8.28 | 1.09 | 0.849 |
| Chloroplast | 5.91a | 0.88b | 2.04b | 0.91b | 1.33b | 0.61 | 0.040 |
| 罗姆布茨菌属Romboutsia | 13.94Aa | 4.96Bb | 5.84Bb | 3.58Bb | 4.02Bb | 0.83 | <0.001 |
| 鞘脂单胞菌属Sphingomonas | 6.66Aa | 7.19Aa | 2.11Bb | 1.79Bb | 2.19Bb | 0.52 | <0.001 |
| 狭义梭菌属1 Clostridium_sensu_stricto_1 | 0.31b | 2.56a | 0.10b | 0.04b | 0.29b | 0.32 | 0.049 |
| 梭菌纲vadinBB60群 Clostridia_vadinBB60_group | 1.56 | 1.66 | 3.20 | 1.60 | 1.95 | 0.25 | 0.202 |
| 拟杆菌属Bacteroides | 2.25 | 3.34 | 3.19 | 3.61 | 4.14 | 0.23 | 0.132 |
| 副拟杆菌属Parabacteroides | 1.10 | 1.33 | 2.58 | 1.55 | 1.45 | 0.47 | 0.211 |
| Gastranaerophilales | 0.28c | 0.88bc | 2.29a | 2.18ab | 1.76ab | 0.45 | 0.010 |
