1 材料与方法
1.1 试验设计和饲粮
表1 饲粮组成及营养水平(干物质基础)Table 1 Composition and nutrient levels of the diet (DM basis)% |
| 项目Items | 含量Content | ||
|---|---|---|---|
| 原料Ingredients | |||
| 玉米Corn | 50.68 | ||
| 豆粕Soybean meal | 24.50 | ||
| 面粉Flour | 10.00 | ||
| 次粉Wheat middling | 7.00 | ||
| 磷酸氢钙CaHPO4 | 1.30 | ||
| 食盐NaCl | 0.30 | ||
| L-赖氨酸硫酸盐L-lysine·H2SO4 | 0.27 | ||
| DL-蛋氨酸DL-methionine | 0.12 | ||
| 石粉Limestone | 1.20 | ||
| 膨润土Bentonite | 3.63 | ||
| 预混料Premix1) | 1.00 | ||
| 合计Total | 100.00 | ||
| 营养水平Nutrient levels2) | |||
| 代谢能Metabolic energy/(MJ/kg) | 11.30 | ||
| 干物质(风干基础) Dry matter (air-dry basis) | 87.30 | ||
| 粗蛋白质Crude protein | 17.00 | ||
| 粗纤维Crude fiber | 3.53 | ||
| 粗脂肪Ether extract | 2.40 | ||
| 钙Calcium | 0.90 | ||
| 总磷Total phosphorus | 0.56 | ||
| 有效磷Available phosphorus | 0.35 | ||
| 赖氨酸Lysine | 0.90 | ||
| 蛋氨酸Methionine | 0.40 | ||
| 蛋氨酸+胱氨酸Methionine+cystine | 0.79 | ||
| 异亮氨酸Isoleucine | 0.73 | ||
| 苏氨酸Threonine | 0.60 | ||
| 色氨酸Tryptophane | 0.26 |
1)预混料为每千克饲粮提供 The premix provided the following per kg of the diet:VA 12 000 IU,VD3 2 500 IU,VE 20 mg,VK3 3 mg,VB1 3 mg,VB2 8 mg,VB6 7 mg,VB12 0.03 mg,生物素 biotin 0.1 mg,泛酸 pantothenic acid 20 mg,烟酸 nicotinic acid 50 mg,叶酸 folic acid 1.5 mg,Cu (as copper sulfate) 9 mg,Zn (as zinc sulfate) 110 mg,Fe (as ferrous sulfate) 100 mg,Mn (as manganese sulfate) 100 mg,Se (as sodium selenite) 0.16 mg,I (as potassium iodide) 0.6 mg。 2)代谢能、有效磷和氨基酸为计算值,其余营养水平为实测值。Metabolic energy, available phosphorus and amino acids were calculated values, while the other nutrient levels were measured values. |
1.2 测定指标及方法
1.2.1 生长性能
1.2.2 屠宰性能
1.2.3 肌肉和盲肠食糜样品采集
1.2.4 胸肌抗氧化指标
1.2.5 肉品质
1.2.6 肌肉蛋白质组学分析
1.2.6.1 蛋白质提取和TMT标记
1.2.6.2 高效液相色谱(high performance liquid chromatograph,HPLC)分馏和液相色谱-串联质谱(liquid chromatography-tandem mass,LC-MS/MS)分析
1.2.6.3 平行反应监测(parallel reaction monitoring,PRM)验证DAPs
1.2.7 肠道菌群分析
1.2.7.1 微生物16S rDNA测序
1.2.7.2 微生物数据分析
1.3 数据统计分析
2 结果与分析
2.1 养殖模式对临武鸭生长性能和屠宰性能的影响
表2 养殖模式对临武鸭生长性能和屠宰性能的影响Table 2 Effects of farming system on growth performance and slaughter performance of Linwu ducks |
| 项目 Items | 组别Groups | 均值标准误 SEM | P值 P-value | |
|---|---|---|---|---|
| FRS | CFS | |||
| 初体重Initial body weight/g | 667.18 | 661.00 | 5.367 | 0.595 |
| 末体重Final body weight/g | 1 485.00 | 1 744.12 | 58.146 | 0.014 |
| 平均日增重Average daily gain/g | 19.47 | 25.79 | 1.437 | 0.016 |
| 平均日采食量Average daily feed intake/g | 268.34 | 268.72 | 0.018 | 0.882 |
| 料重比Feed to gain ratio | 13.79 | 10.63 | 0.621 | 0.002 |
| 屠宰率Carcass rate/% | 86.03 | 84.61 | 0.292 | 0.011 |
| 半净膛率Semi-eviscerated rate/% | 78.57 | 77.79 | 0.237 | 0.100 |
| 全净膛率Eviscerated rate/% | 70.63 | 70.47 | 0.246 | 0.758 |
| 胸肌率Breast muscle rate/% | 12.12 | 13.39 | 0.264 | 0.012 |
| 腿肌率Thigh muscle rate/% | 14.50 | 11.02 | 0.441 | <0.001 |
| 腹脂率Abdominal fat rate/% | 0.87 | 1.68 | 0.121 | <0.001 |
| 心脏指数Heart index/(mg/g) | 7.74 | 6.88 | 0.197 | 0.024 |
| 肝脏指数Liver index/(mg/g) | 20.87 | 16.33 | 0.661 | <0.001 |
| 肌胃指数Gizzard index/(mg/g) | 28.81 | 25.49 | 0.887 | 0.059 |
| 腺胃指数Proventriculus index/(mg/g) | 3.72 | 3.09 | 0.165 | 0.054 |
| 脾脏指数Spleen index/(mg/g) | 0.82 | 0.69 | 0.069 | 0.366 |
2.2 养殖模式对临武鸭胸肌抗氧化指标的影响
表3 养殖模式对临武鸭胸肌抗氧化指标的影响Table 3 Effects of farming system on antioxidant indices in breast muscle of Linwu ducks |
| 项目 Items | 组别Groups | 均值标准误 SEM | P值 P-value | |
|---|---|---|---|---|
| FRS | CFS | |||
| 超氧化物歧化酶SOD/(ng/g) | 8.68 | 7.00 | 0.295 | 0.002 |
| 过氧化氢酶CAT/(pg/g) | 527.40 | 404.00 | 19.298 | <0.001 |
| 谷胱甘肽过氧化物酶GSH-Px/(ng/g) | 13.41 | 9.10 | 0.621 | <0.001 |
| 总抗氧化能力T-AOC/(mmol/g) | 0.37 | 0.31 | 0.010 | 0.001 |
| 丙二醛MDA/(nmol/g) | 7.20 | 7.99 | 0.280 | 0.159 |
2.3 养殖模式对临武鸭肉品质的影响
表4 养殖模式对临武鸭肉品质的影响Table 4 Effects of farming system on meat quality of Linwu ducks |
| 项目 Items | 组别Groups | 均值标准误 SEM | P值 P-value | ||
|---|---|---|---|---|---|
| FRS | CFS | ||||
肉色Meat color | 亮度L* | 40.26 | 53.14 | 1.842 | <0.001 |
| 红度a* | 9.41 | 7.59 | 0.441 | 0.034 | |
| 黄度b* | 2.99 | 5.72 | 0.421 | <0.001 | |
| 剪切力Shear force/N | 49.87 | 22.65 | 3.557 | <0.001 | |
| 滴水损失Drip loss/% | 3.38 | 5.65 | 0.507 | 0.021 | |
2.4 不同养殖模式下临武鸭肌肉蛋白质组学分析
2.4.1 胸肌DAPs及GO功能注释
图1 临武鸭胸肌蛋白质组学分析A:差异积累蛋白;B:DAPs在GO功能注释生物过程中的比例;C:DAPs在GO功能注释细胞组分中的比例;D:DAPs在GO功能注释分子功能中的比例;E:3种GO功能注释下排名前10的术语;F:PRM分析和TMT定量蛋白质组学分析结果比较。A: DAPs;B: proportion of DAPs in biological process of GO functional annotation; C: proportion of DAPs in cell component of GO functional annotation; D: proportion of DAPs in molecular function of GO functional annotation; E: top 10 terms in 3 kinds of GO functional annotation; F: comparison of PRM analysis and TMT quantitative proteomics analysis results. -Log10(p):-log10(P值) -log10(P-value);Log2(Fold Change):log2(差异倍数);Down:下调 down-regulated;NoSig:不显著 non-significant;Up:上调 up-regulated;High:高;Low:低;cellular process:细胞过程;cellular nitrogen compound metabolic process:细胞氮化合物代谢过程;organic substance biosynthetic process:有机物生物合成过程;biosynthetic process:生物合成过程;cellular biosynthetic process:细胞生物合成过程;gene expression:基因表达;cellular nitrogen compound biosynthetic process:细胞氮化合物生物合成过程;organonitrogen compound biosynthetic process:有机氮化合物生物合成过程;macromolecule biosynthetic process:大分子生物合成过程;cellular macromolecule biosynthetic process:细胞大分子生物合成过程;cellular amide metabolic process:细胞酰胺代谢过程;translation:翻译;peptide biosynthetic process:肽生物合成过程;peptide metabolic process:肽代谢过程;amide biosynthetic process:酰胺生物合成过程;positive regulation of cellular process:细胞过程的正调节;cell cycle process:细胞周期过程;protein targeting to mitochondrion:靶向线粒体的蛋白质;protein localization to mitochondrion:定位线粒体的蛋白质;mitochondrial transport:线粒体转运;intracellular anatomical structure:细胞内解剖结构;organelle:细胞器;intracellular organelle:细胞内细胞器;non-membrane-bounded organelle:无膜结合细胞器;intracellular non-membrane-bounded organelle:细胞内无膜结合细胞器;ribosome:核糖体;chromosome:染色体;chromatin:染色质;DNA packaging complex:DNA包装复合物;protein-DNA complex:蛋白质-DNA复合物;nucleosome:核小体;cytosol:胞质溶胶;microtubule:微管;heterocyclic compound binding:杂环化合物结合;organic cyclic compound binding:有机环化合物结合;nucleic acid binding:核酸结合;structural molecule activity:结构分子活性;RNA binding:RNA结合;structural constituent of ribosome:核糖体结构成分;guanyl ribonucleotide binding:脒基核糖核苷酸结合;GTP binding:GTP结合;DNA binding:DNA结合;translation regulator activity:翻译调节活性;structural constituent of chromatin:染色质结构成分;antioxidant activity:抗氧化活性;transferase activity, transferring alkyl or aryl (other than methyl) groups:转移酶活性,转移烷基或芳香基(甲基除外)基团;translation elongation factor activity:翻译延长因子活性;protein-disulfide reductase activity:蛋白质-二硫化物还原酶活性;glutathione transferase activity:谷胱甘肽转移酶活性;disulfide oxidoreductase activity:二硫化物氧化还原酶活性;thioredoxin-disulfide reductase activity:硫氧还蛋白-二硫化物还原酶活性;nucleosomal DNA binding:核小体DNA结合;chromatin DNA binding:染色质DNA结合;Biological Process:生物过程;Cellular Component;细胞组分;Molecular Function:分子功能;POLR2B:RNA聚合酶Ⅱ亚基B RNA polymerase Ⅱ subunit B;PRUNE1:修剪外聚磷酸酶1 prune exopolyphosphatase 1;GTPBP6:GTP结合蛋白6 GTP binding protein 6;ZNF512:锌指蛋白512 zinc finger protein 512;PTMA:前胸腺素α prothymosin α;HMGB3:高迁移率族蛋白B3 high mobility group box 3;Col6a3:Ⅵ型胶原蛋白α3链 collagen type Ⅵ α3 chain;SF3A3:剪接因子3a亚基3 splicing factor 3a subunit 3;Psmc4:蛋白酶体26S亚基ATP酶4 proteasome 26S subunit ATPase 4;Count:计数;log2FC:log2(差异倍数) log2(fold change);TMT:串联质量标签 tandem mass tag;PRM:平行反应监测 parallel reaction monitoring。 Fig.1 Proteomic analysis of breast muscle of Linwu ducks |
2.4.2 DAPs蛋白质互作(protein-protein interaction,PPI)网络及其对肉品质的影响
图2 DAPs蛋白质互作网络限于篇幅,仅注释主要蛋白质名称。Due to space limitations, only names of main proteins were annotated. BPGM:二磷酸甘油酸变位酶 bisphosphoglycerate mutase;PHGDH:磷酸甘油酸脱氢酶 phosphoglycerate dehydrogenase;PSPH:磷酸丝氨酸磷酸酶 phosphoserine phosphatase;LDHA:乳酸脱氢酶A lactate dehydrogenase A;ENOPH1:烯醇酶磷酸酶1 enolase phosphatase 1;BCAT1:支链氨基酸转氨酶1 branched-chain amino acid transaminase 1。图3同 The same as Fig.3。 Fig.2 Protein-protein interaction network of DAPs |
图3 代谢相关DAPs与胸肌肉品质相关指标相关性热图*表示显著相关(P<0.05),**(P<0.01)和***(P<0.001)表示极显著相关。图5同。 Fig.3 Heat map of correlation between metabolism related DAPs and breast muscle quality related indices * indicated significant correlation (P<0.05), and ** (P<0.01) and *** (P<0.001) indicated extremely significant correlation. The same as Fig.5. |
2.5 不同养殖模式下临武鸭肠道菌群分析
2.5.1 肠道菌群结构分析
图4 临武鸭肠道菌群结构分析A:α多样性分析,***表示P<0.001;B:β多样性分析;C:在门水平上的组成;D:在属水平上的组成;E:LEfSe分析。A: α diversity analysis, *** indicated P<0.001; B: β diversity analysis; C: composition at phylum level; D: composition at genus level; E: LEfSe analysis. FRS:FRS组 FRS group;CFS:CFS组 CFS group;PC1:主成分1 principal component 1;PC2:主成分2 principal component 2;Relative Abundance on Phylum Level:门水平相对丰度;Relative Abundance on Genus Level:属水平相对丰度。 仅注释主要菌群的名称 Only names of main microbiota were annotated。Firmicutes:厚壁菌门;Bacteroidota:拟杆菌门;Actinobacteriota:放线菌门;Desulfobacterota:脱硫杆菌门;Deferribacterota:脱铁杆菌门;Proteobacteria:变形菌门;Spirochaetota:螺旋体门;Campilobacterota:弯曲杆菌门;Cyanobacteria:蓝细菌门;Patescibacteria:髌骨菌门;other:其他;Bacteroides:拟杆菌属;Faecalibacterium:粪杆菌属;Rikenellaceae_RC9_gut_group:理研菌科RC9肠道群;Clostridia_UCG-014:梭菌纲UCG-014;Ruminococcus_torques_group:瘤胃球菌属扭链群;Alistipes:另枝菌属;Eubacterium_coprostanoligenes_group:产粪甾醇真细菌群;Desulfovibrio:脱硫弧菌属;Intestinimonas:肠单胞菌属;Mucispirillum:穆齐螺菌属;Phascolarctobacterium:考拉杆菌属。 Fig.4 Analysis of gut microbiota structure of Linwu ducks |

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