1 材料与方法
1.1 伦理声明
1.2 试验设计和饲养管理
表1 试验分组Table 1 Experimental grouping |
| 项目 Item | 饲粮非植酸磷水平Dietary NPP levels/% | |||
|---|---|---|---|---|
| 0.24 | 0.32 | 0.40 | ||
饲粮钙水平 Dietary Ca levels/% | 2.50 | 1组 | 2组 | 3组 |
| 3.00 | 4组 | 5组 | 6组 | |
| 3.50 | 7组 | 8组 | 9组 | |
1.3 检测指标与方法
1.3.1 产蛋性能
1.3.2 蛋品质指标
表2 试验饲粮组成及营养水平(风干基础)Table 2 Composition and nutrient levels of experimental diets (air-dry basis) % |
| 项目 Items | 组别Groups | |||||||||
|---|---|---|---|---|---|---|---|---|---|---|
| 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | ||
| 原料Ingredients | ||||||||||
| 玉米Corn | 65.93 | 65.65 | 66.20 | 62.47 | 63.44 | 63.51 | 61.18 | 61.67 | 61.77 | |
| 小麦麸Wheat bran | 3.00 | 2.50 | 1.75 | 3.00 | 1.95 | 1.58 | 2.00 | 1.05 | 0.65 | |
| 豆粕Soybean meal | 20.18 | 20.08 | 20.21 | 20.52 | 20.68 | 20.79 | 21.24 | 21.33 | 21.42 | |
| 豆油Soybean oil | 0.10 | 0.24 | 0.10 | 1.25 | 1.00 | 1.00 | 1.74 | 1.65 | 1.64 | |
| L-赖氨酸L-Lys | 0.03 | 0.03 | 0.03 | 0.03 | 0.03 | 0.03 | 0.03 | 0.03 | 0.03 | |
| DL-蛋氨酸DL-Met | 0.14 | 0.14 | 0.14 | 0.14 | 0.14 | 0.14 | 0.14 | 0.14 | 0.14 | |
| 磷酸氢钙CaHPO4 | 1.07 | 1.12 | 1.63 | 0.70 | 1.16 | 1.64 | 0.71 | 1.18 | 1.66 | |
| 石粉Limestone | 5.99 | 5.93 | 5.63 | 7.58 | 7.29 | 7.00 | 8.97 | 8.67 | 8.38 | |
| 食盐NaCl | 0.31 | 0.31 | 0.31 | 0.31 | 0.31 | 0.31 | 0.31 | 0.31 | 0.31 | |
| 苜蓿粉Alfalfa meal | 2.25 | 3.00 | 3.00 | 3.00 | 3.00 | 3.00 | 2.68 | 2.97 | 3.00 | |
| 预混料Premix1) | 1.00 | 1.00 | 1.00 | 1.00 | 1.00 | 1.00 | 1.00 | 1.00 | 1.00 | |
| 合计Total | 100.00 | 100.00 | 100.00 | 100.00 | 100.00 | 100.00 | 100.00 | 100.00 | 100.00 | |
| 营养水平Nutrient levels2) | ||||||||||
| 代谢能ME/(MJ/kg) | 11.23 | 11.23 | 11.23 | 11.23 | 11.23 | 11.23 | 11.23 | 11.23 | 11.23 | |
| 粗蛋白质CP | 15.52 | 15.47 | 15.47 | 15.50 | 15.50 | 15.51 | 15.51 | 15.49 | 15.50 | |
| 钙Ca | 2.52 | 2.47 | 2.47 | 2.98 | 3.02 | 2.99 | 3.51 | 3.53 | 3.53 | |
| 总磷TP | 0.51 | 0.60 | 0.68 | 0.52 | 0.59 | 0.67 | 0.51 | 0.59 | 0.67 | |
| 非植酸磷NPP | 0.25 | 0.32 | 0.42 | 0.24 | 0.31 | 0.42 | 0.23 | 0.32 | 0.40 | |
| 蛋氨酸Met | 0.38 | 0.37 | 0.36 | 0.37 | 0.38 | 0.37 | 0.38 | 0.37 | 0.37 | |
| 赖氨酸Lys | 0.78 | 0.77 | 0.77 | 0.75 | 0.76 | 0.78 | 0.75 | 0.77 | 0.78 | |
1)预混料为每千克饲粮提供 The premix provided the following per kg of diets:VA 7 500 IU,VD3 3 250 IU,VE 10 mg,生物素 biotin 0.15 mg,叶酸 folic acid 0.5 mg,甲萘醌 menadione 1 mg,硝酸硫胺 thiamine mononitrate 1.3 mg,核黄素 riboflavin 4.5 mg,吡哆醇 pyridoxine 3 mg,氰钴胺 cyanocobalamin 0.01 mg,烟酰胺 nicotinamide 20 mg,泛酸 pantothenic acid 20 mg,Cu (as copper sulfate) 10 mg,Fe (as ferrous sulfate) 120 mg,Mn (as manganese sulfate) 100 mg,Zn (as zine sulfate) 95 mg。 2)代谢能为计算值,其余为实测值。ME was a calculated value, while the others were measured values. |
1.3.3 肠道消化酶活性
1.3.4 养分表观利用率
1.3.5 蛋壳腺矿化相关基因和Ca转运相关基因表达
表3 基因引物序列Table 3 Primer sequences of genes |
| 基因 Genes | 引物序列 Primer sequences (5'—3') | 登录号 Accession number |
|---|---|---|
| 骨桥蛋白 | F: CAGTGTGGCATATGGCTTCAGG | NM_204535.5 |
| OPN | R: ATCCTCAATGAGCTTCCTGGCA | |
| 钙结合蛋白 | F: TGTACGATCAAGATGGCAATGGA | NM_205513.2 |
| CALB | R: CCATCAGACAAGGCCATGATGC | |
| β-肌动蛋白 | F: TTCTTGGGTATGGAGTCCTGTGGT | NM_205518.2 |
| β-actin | R: CCAGACAGCACTGTGTTGGCATA |
1.4 数据处理与统计分析
2 结果
2.1 饲粮Ca和NPP水平对琅琊鸡产蛋性能的影响
表4 饲粮Ca和NPP水平对产蛋前期琅琊鸡产蛋性能的影响Table 4 Effects of dietary Ca and NPP levels on laying performance of Langya hens during early laying period |
| 项目 Items | 平均日产蛋量 Average daily egg mass/(g/d) | 平均蛋重 Average egg weight/g | 产蛋率 Laying rate/% | 料蛋比 Feed/egg | 平均日采食量 Average daily feed intake/g | |
|---|---|---|---|---|---|---|
组别 Groups | 1 | 18.49a | 39.83 | 46.42a | 4.52d | 83.46 |
| 2 | 17.16b | 40.95 | 41.97b | 4.99cd | 85.64 | |
| 3 | 17.24b | 40.32 | 42.79b | 5.22bc | 90.02 | |
| 4 | 15.91d | 40.92 | 38.85cd | 5.63ab | 89.16 | |
| 5 | 19.13a | 39.86 | 47.99a | 4.80cd | 91.85 | |
| 6 | 16.83bc | 41.45 | 40.69bc | 5.17bc | 86.99 | |
| 7 | 17.03b | 42.01 | 40.55bc | 5.33bc | 90.53 | |
| 8 | 15.25cd | 40.51 | 37.63d | 5.99a | 91.17 | |
| 9 | 17.20b | 41.52 | 41.49b | 5.23bc | 89.97 | |
| SEM | 0.299 | 0.693 | 0.723 | 0.158 | 2.066 | |
| 主效应Main effects | ||||||
钙 Ca/% | 2.50 | 17.63a | 40.36 | 43.72a | 4.91b | 86.37 |
| 3.00 | 17.29a | 40.74 | 42.51a | 5.20b | 89.34 | |
| 3.50 | 16.49b | 41.35 | 39.89b | 5.52a | 90.56 | |
非植酸磷 NPP/% | 0.24 | 17.14 | 40.92 | 41.94 | 5.16 | 87.72 |
| 0.32 | 17.18 | 40.44 | 42.53 | 5.26 | 89.55 | |
| 0.40 | 17.09 | 41.10 | 41.65 | 5.21 | 88.99 | |
| P值P-value | ||||||
| 钙Ca | <0.001 | 0.230 | <0.001 | <0.001 | 0.054 | |
| 非植酸磷NPP | 0.939 | 0.495 | 0.339 | 0.736 | 0.542 | |
| 钙×非植酸磷Ca×NPP | <0.001 | 0.301 | <0.001 | <0.001 | 0.164 |
同列数据肩标不同小写字母表示差异显著(P<0.05),相同或无字母表示差异不显著(P>0.05)。下表同。 | |
In the same column, values with different small letter superscripts mean significant difference (P<0.05), while with the same or no letter superscripts mean no significant difference (P>0.05). The same as below. |
2.2 饲粮Ca和NPP水平对琅琊鸡蛋品质的影响
表5 饲粮Ca和NPP水平对产蛋前期琅琊鸡蛋品质的影响Table 5 Effects of dietary Ca and NPP levels on egg quality of Langya hens during early laying period |
| 项目 Items | 蛋重 Egg weight/g | 蛋白高度 Albumen height/mm | 蛋黄颜色 Yolk color | 哈氏单位 Haugh unit | 蛋壳强度 Eggshell strength/N | 蛋形指数 Egg shape index | 蛋壳厚度 Eggshell thickness/ mm | ||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
组别 Groups | 1 | 42.72 | 5.05ab | 8.00 | 75.23cd | 3.45c | 1.28 | 0.39 | |||||||||
| 2 | 44.55 | 5.33ab | 8.20 | 78.41bc | 3.64c | 1.30 | 0.38 | ||||||||||
| 3 | 45.48 | 4.91ab | 8.00 | 80.20ab | 4.15ab | 1.32 | 0.41 | ||||||||||
| 4 | 44.37 | 4.86ab | 8.20 | 79.25abc | 4.03b | 1.29 | 0.40 | ||||||||||
| 5 | 44.50 | 4.64b | 8.00 | 78.37bc | 4.19ab | 1.29 | 0.39 | ||||||||||
| 6 | 45.08 | 5.77a | 8.20 | 82.76a | 4.14ab | 1.29 | 0.38 | ||||||||||
| 7 | 43.01 | 5.21ab | 8.10 | 77.85bc | 4.02b | 1.29 | 0.41 | ||||||||||
| 8 | 44.88 | 5.03ab | 8.00 | 79.30abc | 4.18ab | 1.27 | 0.38 | ||||||||||
| 9 | 42.37 | 4.53b | 8.10 | 73.77d | 4.31a | 1.30 | 0.40 | ||||||||||
| SEM | 0.776 | 0.314 | 0.256 | 1.306 | 0.087 | 0.015 | 0.011 | ||||||||||
| 主效应Main effects | |||||||||||||||||
钙 Ca/% | 2.50 | 44.25 | 5.10 | 8.07 | 77.95b | 3.75b | 1.30 | 0.39 | |||||||||
| 3.00 | 44.65 | 5.09 | 8.13 | 80.13a | 4.12a | 1.29 | 0.39 | ||||||||||
| 3.50 | 43.42 | 4.93 | 8.07 | 76.98b | 4.17a | 1.29 | 0.40 | ||||||||||
非植酸磷 NPP/% | 0.24 | 43.37 | 5.04 | 8.10 | 77.45 | 3.83c | 1.29 | 0.40 | |||||||||
| 0.32 | 44.65 | 5.00 | 8.07 | 78.70 | 4.00b | 1.29 | 0.39 | ||||||||||
| 0.40 | 44.31 | 5.08 | 8.10 | 78.91 | 4.20a | 1.30 | 0.40 | ||||||||||
| P值P-value | |||||||||||||||||
| 钙Ca | 0.154 | 0.750 | 0.935 | 0.017 | <0.001 | 0.527 | 0.670 | ||||||||||
| 非植酸磷NPP | 0.127 | 0.960 | 0.983 | 0.343 | <0.001 | 0.356 | 0.380 | ||||||||||
| 钙×非植酸磷Ca×NPP | 0.103 | 0.048 | 0.925 | 0.002 | 0.011 | 0.514 | 0.244 | ||||||||||
2.3 饲粮Ca和NPP水平对产蛋前期琅琊鸡肠道消化酶活性的影响
表6 饲粮Ca和NPP水平对产蛋前期琅琊鸡肠道消化酶活性的影响Table 6 Effects of dietary Ca and NPP levels on intestinal digestive enzyme activities of Langya hens during early laying period U/mg prot |
| 项目 Items | 脂肪酶 Lipase | 胰蛋白酶 Trypsin | 淀粉酶 Amylase | |
|---|---|---|---|---|
组别 Groups | 1 | 163.96 | 2 458.38c | 7.76 |
| 2 | 164.12 | 2 522.39b | 7.75 | |
| 3 | 164.91 | 2 446.19cd | 7.79 | |
| 4 | 164.51 | 2 518.69b | 7.87 | |
| 5 | 165.30 | 2 640.24a | 7.99 | |
| 6 | 165.21 | 2 512.46b | 7.59 | |
| 7 | 162.89 | 2 406.87d | 7.52 | |
| 8 | 164.91 | 2 515.78b | 7.54 | |
| 9 | 163.63 | 2 407.96d | 7.27 | |
| SEM | 1.751 | 9.600 | 0.209 | |
| 主效应Main effects | ||||
钙 Ca/% | 2.50 | 164.33 | 2 475.66b | 7.77 |
| 3.00 | 165.01 | 2 557.13a | 7.82 | |
| 3.50 | 163.81 | 2 443.54c | 7.44 | |
非植酸磷 NPP/% | 0.24 | 163.79 | 2 461.31b | 7.72 |
| 0.32 | 164.78 | 2 559.47a | 7.76 | |
| 0.40 | 164.59 | 2 455.54b | 7.55 | |
| P值P-value | ||||
| 钙Ca | 0.705 | <0.001 | 0.071 | |
| 非植酸磷NPP | 0.764 | <0.001 | 0.434 | |
| 钙×非植酸磷Ca×NPP | 0.979 | 0.033 | 0.852 |
2.4 饲粮Ca和NPP水平对产蛋前期琅琊鸡养分表观利用率的影响
表7 饲粮Ca和NPP水平对产蛋前期琅琊鸡养分表观利用率的影响Table 7 Effects of dietary Ca and NPP levels on nutrient apparent utilization rates of Langya hens during early laying period % |
| 项目 Items | 干物质 DM | 粗脂肪 EE | 粗蛋白质 CP | 粗灰分 Ash | 钙 Ca | 磷 P | |
|---|---|---|---|---|---|---|---|
组别 Groups | 1 | 75.85 | 73.30 | 53.99 | 36.77 | 48.83bc | 39.34c |
| 2 | 75.76 | 74.12 | 55.22 | 37.14 | 49.65abc | 41.06abc | |
| 3 | 75.60 | 73.57 | 53.39 | 36.47 | 47.67c | 39.31c | |
| 4 | 75.19 | 74.32 | 55.22 | 37.19 | 48.03bc | 39.90bc | |
| 5 | 76.02 | 74.71 | 56.23 | 38.34 | 51.14a | 42.66a | |
| 6 | 75.59 | 73.08 | 54.29 | 37.03 | 50.03ab | 42.13ab | |
| 7 | 75.33 | 73.93 | 53.07 | 37.30 | 49.09abc | 40.86abc | |
| 8 | 75.09 | 74.28 | 54.64 | 37.60 | 48.01bc | 38.92c | |
| 9 | 75.40 | 73.81 | 53.80 | 37.54 | 47.57c | 39.48c | |
| SEM | 0.504 | 0.621 | 0.675 | 0.604 | 0.672 | 0.779 | |
| 主效应Main effects | |||||||
钙 Ca/% | 2.50 | 75.74 | 73.66 | 54.20ab | 36.79 | 48.72ab | 39.90b |
| 3.00 | 75.60 | 74.04 | 55.25a | 37.52 | 49.74a | 41.56a | |
| 3.50 | 75.27 | 74.01 | 53.83b | 37.48 | 48.23b | 39.75b | |
非植酸磷 NPP/% | 0.24 | 75.46 | 73.85 | 54.09b | 37.09 | 48.65 | 40.03 |
| 0.32 | 75.62 | 74.37 | 55.36a | 37.69 | 49.60 | 40.88 | |
| 0.40 | 75.53 | 73.49 | 53.83b | 37.01 | 48.43 | 40.30 | |
| P值P-value | |||||||
| 钙Ca | 0.470 | 0.716 | 0.040 | 0.203 | 0.028 | 0.012 | |
| 非植酸磷NPP | 0.910 | 0.232 | 0.019 | 0.262 | 0.088 | 0.406 | |
| 钙×非植酸磷Ca×NPP | 0.780 | 0.755 | 0.774 | 0.820 | 0.023 | 0.033 |
2.5 饲粮Ca和NPP水平对产蛋前期琅琊鸡蛋壳腺OPN和CALB表达的影响
表8 饲粮Ca和NPP水平对产蛋前期琅琊鸡蛋壳腺OPN和CALB表达的影响Table 8 Effects of dietary Ca and NPP levels on expression of OPN and CALB in eggshell gland of Langya hens during early laying period |
| 项目Items | 骨桥蛋白OPN | 钙结合蛋白CALB | |
|---|---|---|---|
组别 Groups | 1 | 1.00d | 1.00f |
| 2 | 1.39ab | 1.13e | |
| 3 | 1.43ab | 1.25d | |
| 4 | 1.10cd | 1.39c | |
| 5 | 1.48a | 1.54ab | |
| 6 | 1.34b | 1.45bc | |
| 7 | 1.14c | 1.45bc | |
| 8 | 1.34b | 1.49abc | |
| 9 | 1.47a | 1.56a | |
| SEM | 0.022 | 0.023 | |
| 主效应Main effects | |||
钙 Ca/% | 2.50 | 1.28 | 1.13b |
| 3.00 | 1.30 | 1.46a | |
| 3.50 | 1.32 | 1.50a | |
非植酸磷 NPP/% | 0.24 | 1.08b | 1.28b |
| 0.32 | 1.40a | 1.39a | |
| 0.40 | 1.41a | 1.42a | |
| P值P-value | |||
| 钙Ca | 0.059 | <0.001 | |
| 非植酸磷NPP | <0.001 | <0.001 | |
| 钙×非植酸磷Ca×NPP | <0.001 | <0.001 |

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