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(风干基础)Table 1 Composition and nutrient levels of experimental diets (air-dry basis)% |
| 项目 Items | 第1~12天 Days 1 to 12 | 第13~55天 Days 13 to 55 | |||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| CON组 CON group | UD组 UD group | LUD组 LUD group | CON组 CON group | UD组 UD group | LUD组 LUD group | ||||||||||
| 原料 Ingredients | |||||||||||||||
| 玉米 Cron | 65.12 | 44.66 | 50.15 | 70.59 | 30.85 | 37.36 | |||||||||
| 小麦 Wheat grain | 6.00 | 6.00 | 12.00 | 12.00 | |||||||||||
| 大麦 Barley | 5.20 | 5.20 | 6.00 | 6.00 | |||||||||||
| 高粱 Sorghum grain | 5.00 | 5.00 | 10.00 | 10.00 | |||||||||||
| 糙米 Brain rice | 5.50 | 5.50 | 12.50 | 12.50 | |||||||||||
| 豆粕 Soybean meal | 26.00 | 20.06 | 13.57 | 20.53 | 11.20 | 4.82 | |||||||||
| 菜籽粕 Rapeseed meal | 1.50 | 1.50 | 2.50 | 2.50 | |||||||||||
| 棉籽粕 Cottonseed meal | 1.50 | 1.50 | 1.50 | 1.50 | |||||||||||
| 亚麻籽粕 Flaxseed meal | 1.50 | 1.50 | 2.50 | 2.50 | |||||||||||
| 玉米干酒糟及其可溶物 Corn DDGS | 1.50 | 1.50 | 3.00 | 3.00 | |||||||||||
| 小麦麸 Wheat bran | 2.00 | 1.00 | 2.00 | ||||||||||||
| 大豆油 Soybean oil | 0.60 | 1.20 | 0.80 | 0.95 | 1.90 | 1.35 | |||||||||
| 鱼粉 Fish meal | 1.50 | 1.50 | 1.50 | 1.50 | 1.50 | 1.50 | |||||||||
| 蔗糖 Sucrose | 1.50 | 1.50 | 1.50 | 1.50 | 1.50 | 1.50 | |||||||||
| 氯化钠 NaCl | 0.20 | 0.20 | 0.20 | 0.20 | 0.20 | 0.20 | |||||||||
| 磷酸氢钙 CaHPO4 | 1.00 | 0.95 | 1.00 | 0.90 | 0.80 | 0.85 | |||||||||
| 碳酸钙 CaCO3 | 0.87 | 0.87 | 0.90 | 0.90 | 0.90 | 0.94 | |||||||||
| L-赖氨酸盐酸盐 L-Lys·HCl | 0.44 | 0.56 | 0.75 | 0.30 | 0.47 | 0.67 | |||||||||
| DL-蛋氨酸 DL-Met | 0.08 | 0.09 | 0.11 | 0.02 | 0.03 | 0.06 | |||||||||
| L-苏氨酸 L-Thr | 0.15 | 0.17 | 0.26 | 0.08 | 0.12 | 0.20 | |||||||||
| L-色氨酸 L-Trp | 0.04 | 0.04 | 0.06 | 0.03 | 0.03 | 0.05 | |||||||||
| 氯化胆碱 Choline chloride | 0.15 | 0.15 | 0.15 | 0.15 | 0.15 | 0.15 | |||||||||
| 预混料 Premix1) | 0.35 | 0.35 | 0.35 | 0.35 | 0.35 | 0.35 | |||||||||
| 合计 Total | 100.00 | 100.00 | 100.00 | 100.00 | 100.00 | 100.00 | |||||||||
| 营养水平 Nutrient levels2) | |||||||||||||||
| 总能 GE/(MJ/kg) | 15.69 | 15.77 | 15.69 | 15.86 | 15.98 | 16.23 | |||||||||
| 代谢能 ME/(MJ/kg) | 13.09 | 12.97 | 12.89 | 13.30 | 13.01 | 12.93 | |||||||||
| 净能 NE/(MJ/kg) | 10.08 | 10.08 | 10.08 | 10.33 | 10.33 | 10.33 | |||||||||
| 粗蛋白质 CP | 18.94 | 18.88 | 16.58 | 15.75 | 16.28 | 14.15 | |||||||||
| 钙 Ca | 0.70 | 0.70 | 0.70 | 0.67 | 0.67 | 0.67 | |||||||||
| 有效磷 AP | 0.33 | 0.33 | 0.33 | 0.31 | 0.31 | 0.31 | |||||||||
| 粗纤维 CF | 2.13 | 2.17 | 2.95 | 2.25 | 2.66 | 2.30 | |||||||||
| 可溶性纤维 SDF | 1.66 | 2.01 | 1.98 | 1.61 | 2.06 | 2.01 | |||||||||
| 不可溶性纤维 IDF | 11.65 | 12.12 | 12.00 | 11.33 | 12.43 | 12.07 | |||||||||
| 总膳食纤维 TDF | 13.31 | 14.14 | 13.99 | 12.85 | 14.29 | 14.18 | |||||||||
| 标准回肠可消化赖氨酸 SID Lys | 1.23 | 1.23 | 1.23 | 0.98 | 0.98 | 0.98 | |||||||||
| 标准回肠可消化蛋氨酸 SID Met | 0.36 | 0.36 | 0.36 | 0.28 | 0.28 | 0.28 | |||||||||
| 标准回肠可消化苏氨酸 SID Thr | 0.73 | 0.73 | 0.73 | 0.59 | 0.59 | 0.59 | |||||||||
| 标准回肠可消化色氨酸 SID Trp | 0.20 | 0.20 | 0.20 | 0.17 | 0.17 | 0.17 | |||||||||
1)预混料为每千克饲粮提供 The premix provided the following per kg of diets:VA 12 000 U,VB1 2 mg,VB2 7.2 mg,VB5 25 mg,VB12 0.02 mg,VD3 2 300 U,VE 100 U,VK3 4.8 mg,生物素 biotin 0.48 mg,叶酸 folic acid 4.0 mg,烟酸 nicotinic acid 40 mg,Cu (as copper sulfate) 5 mg,Fe (as ferrous sulfate) 100 mg,Mn (as manganese sulfate) 3 mg,Zn (as zinc sulfate) 80 mg,I (as potassium iodide) 0.14 mg,Se (as sodium selenite) 0.25 mg。 | |
2)总能、粗蛋白质和粗纤维为实测值,其余为计算值,均根据《中国饲料成分及营养价值表(2022年第33版)》计算。GE, CP and CF were measured values, while the others were calculated values, all calculated with reference to the Tables of Feed Composition and Nutritive Values in China (33rd edition, 2022). |
1.3 样品采集
1.4 指标测定
1.4.1 生长性能
1.4.2 营养物质含量和表观消化率
无氮浸出物含量(%)=干物质含量-粗蛋白质
含量-粗脂肪含量-粗纤维含量-粗灰分含量;
有机物含量(%)=干物质含量-粗灰分含量;
某营养物质表观消化率(%)=[1-(饲粮中指示剂
含量/粪便中指示剂含量)×(粪便中
该营养物质含量/饲粮中
该营养物质含量)]×100。
1.4.3 血清生化指标以及血清和睾丸组织的抗氧化与激素指标
1.4.4 睾丸发育指标
睾丸体积(cm3)=0.523 6×长径×短径2;
睾丸指数(%)=睾丸重量/体重。
1.4.5 总RNA的提取和实时荧光定量PCR(qRT-PCR)
表2 引物序列Table 2 Primer sequences |
| 基因 Genes | 引物序列 Primer sequences (5'—3') | |
|---|---|---|
| 细胞周期蛋白B1 CCNB1 | F:TGGTGAATGGACACCAACTC R:TGCTTTGTAAGCCCTCGATT | |
| 细胞周期蛋白依赖性激酶4 CDK4 | F:GCATCCCAATGTTGTCCG R:GGGGTGCCTTGTCCAGATA | |
| 类固醇合成急性调节蛋白 STAR | F:GGTTCTCAGCTGGAAGACACT R:ACCTCGTCCCCATTCTCCTG | |
| 细胞色素P450 11A1 P450scc | F:TCGGCAACTTGGAATCTGTT R:AATGCTGGTGATAGGCAACC | |
| 细胞色素P450 17A1 CYP17A1 | F:GACTACCGTGGTAATTGGTGAC R:GCAATCCCCTTTTGGTTGTC | |
| 17β-羟基类固醇脱氢酶 17β-HSD | F:ACTTGGCTGTTCGCCTAGC R:GAGGGCATCCTTGAGTCCTG | |
| β-肌动蛋白 β-actin | F:TCTGGCACCACACCTTCT R:TGATCTGGGTCATCTTCTCAC | |
1.5 数据统计分析
2 结果
2.1 杂粮杂粕和低蛋白质杂粮杂粕对后备公猪生长性能的影响
表3 杂粮杂粕饲粮和低蛋白质杂粮杂粕饲粮对后备公猪生长性能的影响Table 3 Effects of miscellaneous grain and meal and low-protein miscellaneous grain and meal diets on growth performance of replacement boars |
| 项目 Items | CON组 CON group | UD组 UD group | LUD组 LUD group | P1值 P1-value | P2值 P2-value | |
|---|---|---|---|---|---|---|
| 前期 Early phase | ||||||
| 初始体重 Initial weight/kg | 17.61±0.40 | 17.88±0.51 | 17.78±0.41 | 0.680 | 0.993 | |
| 结束体重 Final weight/kg | 26.33±0.50 | 27.94±0.67 | 27.33±0.56 | 0.073 | 0.263 | |
| 平均日采食量 ADFI/(g/d) | 1 200.38±18.48 | 1 218.24±22.75 | 1 225.63±25.46 | 0.551 | 0.574 | |
| 平均日增重 ADG/(g/d) | 696.15±23.01 | 758.65±19.13 | 716.83±20.43 | 0.055 | 0.056 | |
| 料重比 F/G | 1.75±0.04 | 1.60±0.03 | 1.70±0.03 | 0.014 | 0.014 | |
| 中期 Middle phase | ||||||
| 初始体重 Initial weight/kg | 26.33±0.50 | 27.94±0.67 | 27.33±0.56 | 0.073 | 0.263 | |
| 结束体重 Final weight/kg | 47.50±1.09 | 48.10±0.99 | 45.72±1.34 | 0.690 | 0.172 | |
| 平均日采食量 ADFI/(g/d) | 2 037.19±57.25 | 1 968.05±55.45 | 1 841.55±91.12 | 0.398 | 0.257 | |
| 平均日增重 ADG/(g/d) | 962.12±32.58 | 916.16±23.86 | 835.86±43.65 | 0.272 | 0.132 | |
| 料重比 F/G | 2.12±0.02 | 2.15±0.05 | 2.21±0.04 | 0.572 | 0.379 | |
| 后期 Late phase | ||||||
| 初始体重 Initial weight/kg | 47.50±1.09 | 48.10±0.99 | 45.72±1.34 | 0.690 | 0.172 | |
| 结束体重 Final weight/kg | 74.00±1.43 | 73.39±1.47 | 69.06±2.21 | 0.769 | 0.123 | |
| 平均日采食量 ADFI/(g/d) | 2 695.06±91.12 | 2 651.35±96.12 | 2 473.79±147.09 | 0.746 | 0.327 | |
| 平均日增重 ADG/(g/d) | 1 204.55±29.83 | 1 149.49±39.15 | 1 060.61±51.10 | 0.280 | 0.186 | |
| 料重比 F/G | 2.24±0.04 | 2.31±0.05 | 2.32±0.05 | 0.274 | 0.848 | |
| 全期 Overall phase | ||||||
| 初始体重 Initial weight/kg | 17.61±0.40 | 17.88±0.51 | 17.78±0.41 | 0.680 | 0.993 | |
| 结束体重 Final weight/kg | 74.00±1.43 | 73.39±1.47 | 69.06±2.21 | 0.769 | 0.123 | |
| 平均日采食量 ADFI/(g/d) | 2 089.63±56.15 | 2 050.22±50.77 | 1 935.66±93.16 | 0.610 | 0.301 | |
| 平均日增重 ADG/(g/d) | 1 006.94±24.55 | 991.17±24.59 | 915.67±37.07 | 0.656 | 0.112 | |
| 料重比 F/G | 2.07±0.02 | 2.07±0.03 | 2.11±0.03 | 0.917 | 0.395 | |
P1:CON组vs UD组 CON group vs UD group;P2:UD组vs LUD组 UD group vs LUD group。下表同the same as below。 |
2.2 杂粮杂粕和低蛋白质杂粮杂粕对后备公猪营养物质表观消化率的影响
表4 杂粮杂粕和低蛋白质杂粮杂粕饲粮对后备公猪营养物质表观消化率的影响Table 4 Effects of miscellaneous grain and meal and low-protein miscellaneous grain and meal diets on nutrient apparent digestibility of replacement boars% |
| 项目 Items | CON组 CON group | UD组 UD group | LUD组 LUD group | P1值 P1-value | P2值 P2-value |
|---|---|---|---|---|---|
| 中期 Middle phase | |||||
| 干物质 DM | 85.29±0.44 | 84.01±0.36 | 81.14±0.46 | 0.040 | <0.001 |
| 粗蛋白质 CP | 80.04±1.18 | 76.19±0.56 | 70.16±1.11 | 0.009 | <0.001 |
| 粗脂肪 EE | 57.65±1.18 | 62.55±1.25 | 59.80±1.32 | 0.012 | <0.001 |
| 粗纤维 CF | 43.88±2.92 | 45.66±1.42 | 26.13±1.75 | 0.590 | <0.001 |
| 总能 GE | 86.97±0.40 | 85.80±0.34 | 83.50±0.41 | 0.042 | <0.001 |
| 有机物 OM | 86.93±0.44 | 85.97±0.38 | 84.04±0.41 | 0.114 | 0.003 |
| 无氮浸出物 NFE | 92.33±0.18 | 92.43±0.32 | 91.55±0.24 | 0.800 | 0.042 |
| 后期 Late phase | |||||
| 干物质 DM | 86.50±0.42 | 85.44±0.42 | 83.65±0.58 | 0.090 | 0.023 |
| 粗蛋白质 CP | 82.10±1.05 | 79.80±0.81 | 75.35±1.06 | 0.102 | <0.001 |
| 粗脂肪 EE | 63.05±0.65 | 64.99±1.21 | 67.08±0.75 | 0.174 | 0.161 |
| 粗纤维 CF | 48.97±2.64 | 46.67±2.80 | 32.56±2.13 | 0.557 | 0.001 |
| 总能 GE | 88.59±0.39 | 87.20±0.36 | 85.85±0.54 | 0.019 | 0.052 |
| 有机物 OM | 88.62±0.41 | 87.49±0.39 | 85.90±0.51 | 0.064 | 0.026 |
| 无氮浸出物 NFE | 93.62±0.24 | 93.34±0.22 | 92.00±0.39 | 0.397 | 0.009 |
2.3 杂粮杂粕和低蛋白质杂粮杂粕对后备公猪血清生化指标及血清和睾丸抗氧化指标的影响
表5 杂粮杂粕和低蛋白质杂粮杂粕饲粮对后备公猪血清生化指标的影响Table 5 Effects of miscellaneous grain and meal and low-protein miscellaneous grain and meal diets on the serum biochemistry indexes of replacement boars |
| 项目 Items | CON组 CON group | UD组 UD group | LUD组 LUD group | P1值 P1-value | P2值 P2-value |
|---|---|---|---|---|---|
| 葡萄糖 GLU/(mmol/L) | 6.05±0.21 | 5.85±0.12 | 6.33±0.20 | 0.412 | 0.053 |
| 总蛋白 TP/(g/L) | 54.75±1.23 | 53.74±0.52 | 51.73±0.83 | 0.459 | 0.056 |
| 总胆固醇 TC/(mmol/L) | 1.93±0.09 | 2.16±0.10 | 2.24±0.08 | 0.101 | 0.570 |
| 甘油三酯 TG/(mmol/L) | 0.43±0.04 | 0.41±0.04 | 0.44±0.03 | 0.789 | 0.613 |
| 低密度脂蛋白胆固醇 LDL-C/(mmol/L) | 1.00±0.05 | 0.92±0.06 | 0.98±0.03 | 0.277 | 0.339 |
| 高密度脂蛋白胆固醇 HDL-C/(mmol/L) | 0.59±0.02 | 0.60±0.04 | 0.59±0.03 | 0.669 | 0.828 |
| 尿素氮 UN/(mmol/L) | 2.71±0.22 | 2.23±0.18 | 1.23±0.16 | 0.104 | 0.001 |
| 乳酸脱氢酶 LDH/(U/L) | 675.77±47.95 | 592.38±28.36 | 658.97±38.46 | 0.154 | 0.183 |
| 谷丙转氨酶 ALT/(U/L) | 112.10±7.18 | 101.35±6.86 | 106.44±5.09 | 0.297 | 0.559 |
| 谷草转氨酶 AST/(U/L) | 30.38±3.35 | 29.02±3.41 | 26.00±2.55 | 0.559 | 0.489 |
表6 杂粮杂粕和低蛋白质杂粮杂粕饲粮对后备公猪血清和睾丸抗氧化指标的影响Table 6 Effects of miscellaneous grain and meal and low-protein miscellaneous grain and meal diets on serum and testicular antioxidant indexes of replacement boars |
| 项目 Items | CON组 CON group | UD组 UD group | LUD组 LUD group | P1值 P1-value | P2值 P2-value |
|---|---|---|---|---|---|
| 血清 Serum | |||||
| 超氧化物歧化酶 SOD/(U/mL) | 4.44±0.44 | 4.54±0.45 | 5.09±0.40 | 0.880 | 0.373 |
| 丙二醛 MDA/(nmol/mL) | 1.17±0.21 | 1.29±0.05 | 1.47±0.18 | 0.764 | 0.352 |
| 过氧化氢酶 CAT/(U/mL) | 143.86±4.68 | 154.69±3.63 | 143.77±5.55 | 0.566 | 0.126 |
| 总抗氧化能力 T-AOC/(μmol/mL) | 3.87±0.11 | 4.03±0.07 | 4.02±0.05 | 0.263 | 0.992 |
| 睾丸 Testis | |||||
| 超氧化物歧化酶 SOD/(U/mg) | 12.87±0.59 | 11.70±1.61 | 12.71±1.36 | 0.508 | 0.640 |
| 丙二醛 MDA/(nmol/mg) | 10.75±2.67 | 7.78±1.52 | 12.07±2.22 | 0.326 | 0.137 |
| 过氧化氢酶 CAT/(U/mg) | 10.38±1.81 | 8.33±2.25 | 5.26±1.02 | 0.256 | 0.266 |
| 总抗氧化能力 T-AOC/(μmol/mg) | 3.68±0.10 | 3.45±0.14 | 3.66±0.11 | 0.224 | 0.266 |
2.4 杂粮杂粕和低蛋白质杂粮杂粕对后备公猪睾丸发育和激素指标的影响
2.4.1 睾丸体积和睾丸指数
表7 杂粮杂粕和低蛋白质杂粮杂粕饲粮对后备公猪睾丸体积和睾丸指数的影响Table 7 Effects of miscellaneous grain and meal and low-protein miscellaneous grain and meal diets on testicular volume and index of replacement boars |
| 项目 Items | CON组 CON group | UD组 UD group | LUD组 LUD group | P1值 P1-value | P2值 P2-value |
|---|---|---|---|---|---|
| 睾丸体积 Testicular volume/cm3 | 51.07±2.93 | 51.60±6.16 | 49.19±4.78 | 0.939 | 0.762 |
| 睾丸重量 Testicular weight/g | 106.89±8.21 | 110.33±9.32 | 102.00±10.16 | 0.785 | 0.554 |
| 睾丸指数 Testicular index/% | 0.14±0.01 | 0.15±0.01 | 0.15±0.02 | 0.700 | 0.918 |
| 附睾重量 Epididymal weight/g | 22.22±0.63 | 25.83±1.87 | 22.56±0.58 | 0.769 | 0.476 |
| 附睾长 Epididymis length/mm | 135.25±3.15 | 132.30±9.37 | 139.52±3.22 | 0.086 | 0.113 |
2.4.2 睾丸组织形态学指标
图1 120日龄后备公猪睾丸组织切片结果Fig.1 Histological section results of testicular tissues from 120-day-old replacement boars A、D:CON组 CON group;B、E:UD组 UD group;C、F:LUD组 LUD group。 A、B、C:100×;D、E、F:400×。 |
表8 杂粮杂粕和低蛋白质杂粮杂粕饲粮对后备公猪睾丸组织学形态指标的影响Table 8 Effects of miscellaneous grain and meal and low-protein miscellaneous grain and meal diets on histological morphology indexes of testicular tissue of replacement boars |
| 项目 Items | CON组 CON group | UD组 UD group | LUD组 LUD group | P1值 P1-value | P2值 P2-value |
|---|---|---|---|---|---|
| 每管生精小管中精原细胞数量 Number of spermatogonia per seminiferous tubule/个 | 22.20±1.12 | 25.48±1.33 | 24.40±1.54 | 0.096 | 0.610 |
| 每管生精小管中支持细胞数量 Number of Sertoli cells per seminiferous tubule/个 | 10.08±0.79 | 10.92±1.64 | 8.16±1.55 | 0.657 | 0.256 |
| 生精小管直径 Seminiferous tubular diameter/μm | 126.62±2.89 | 133.63±6.79 | 130.96±8.84 | 0.370 | 0.817 |
| 生精上皮厚度 Seminiferous epithelium height/μm | 39.66±1.28 | 37.76±1.44 | 37.90±2.52 | 0.353 | 0.962 |
2.4.3 杂粮杂粕和低蛋白质杂粮杂粕饲粮对后备公猪血清和睾丸激素指标的影响
表9 杂粮杂粕和低蛋白质杂粮杂粕饲粮对后备公猪血清和睾丸激素指标的影响Table 9 Effects of miscellaneous grain and meal and low-protein miscellaneous grain and meal diets on serum and testicular hormone indexes of replacement boars |
| 项目 Items | CON组 CON group | UD组 UD group | LUD组 LUD group | P1值 P1-value | P2值 P2-value |
|---|---|---|---|---|---|
| 血清 Serum | |||||
| 促卵泡素 FSH/(mU/mL) | 33.52±2.50 | 30.30±4.92 | 29.82±4.98 | 0.554 | 0.948 |
| 黄体生成素 LH/(mU/mL) | 29.26±0.22 | 30.21±0.29 | 30.00±0.23 | 0.022 | 0.568 |
| 睾酮 T/(pg/mL) | 60.28±7.39 | 66.43±7.70 | 58.21±7.26 | 0.577 | 0.455 |
| 睾丸 Testis | |||||
| 促卵泡素 FSH/(mU/mg) | 3.50±0.30 | 2.71±0.28 | 1.91±0.15 | 0.076 | 0.028 |
| 黄体生成素 LH/(mU/mg) | 3.72±0.45 | 3.84±0.44 | 3.96±0.26 | 0.856 | 0.718 |
| 睾酮 T/(pg/mg) | 370.35±8.27 | 381.05±11.91 | 396.11±8.23 | 0.462 | 0.329 |
2.5 杂粮杂粕和低蛋白质杂粮杂粕饲粮对后备公猪睾丸细胞增殖和睾酮合成相关基因表达的影响
图2 杂粮杂粕和低蛋白质杂粮杂粕饲粮对后备公猪睾丸细胞增殖和睾酮合成相关基因表达的影响“*”表示差异显著(P<0.05)。“*” indicated a significant difference (P<0.05). CDK4:细胞周期蛋白依赖性激酶 4;CCNB1:细胞周期蛋白 B1;STAR:类固醇合成急性调节蛋白;P450scc:细胞色素 P450 11A1;CYP17A1:细胞色素 P450 17A1;17β-HSD:17β- 羟基类固醇脱氢酶。 Fig.2 Effects of miscellaneous grain and meal and low-protein miscellaneous grain and meal diets on expression of genes related to testicular cell proliferation and testosterone synthesis of replacement boars |
