Chinese Journal of Animal Nutrition >

2024 , Vol. 36 >Issue 5: 2992 - 3002

DOI: https://doi.org/10.12418/CJAN2024.257

RESEARCH PAPER

Effects of Substituting 25-Hydroxyvitamin D3 for Vitamin D3 on Performance, Egg Quality and Calcium and Phosphorus Metabolism of Laying Hens in Late Laying Period

  • GAO Shan , 1 ,
  • ZHENG Junjie 2 ,
  • QIU Kai 1 ,
  • ZHANG Haijun 1 ,
  • WU Shugeng , 1, *
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  • 1 Laboratory of Quality & Safety Risk Assessment for Animal Products on Feed Hazards, Key Laboratory of Feed Biotechnology, Ministry of Agriculture and Rural Affairs, Institute of Feed Research, Chinese Academy of Agricultural Sciences, Beijing 100081, China
  • 2 Beijing Agricultural Products Quality and Safety Center, Beijing 100020, China
* professor, E-mail:

Received date: 2023-11-29

  Online published: 2024-05-15

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Abstract

This experiment was conducted to investigate the effects of dietary 25-hydroxyvitamin D3 (25-OHD3) as a substitute for vitamin D3 (VD3) on performance, egg quality, calcium and phosphorus metabolism, intestinal morphology and tibia quality of laying hens in late laying period. A total of 270 Hy-Line brown laying hens aged at 55 weeks with normal feeding and similar body weight (uniformity>90%) were randomly divided into 3 groups with 6 replicates per group and 15 hens per replicate. The hens in the control group were fed a basal diet supplemented with 4 000 IU/kg VD3, the hens in experimental group 1 were fed a diet supplemented with 50 μg/kg 25-OHD3 and 2 000 IU/kg VD3, and the hens in experimental group 2 were fed a diet supplemented with 100 μg/kg 25-OHD3. The pre-trial period was 1 week and the experimental period was 12 weeks. The results showed as follows: 1) compared with the control group, the laying rate in experimental group 1 in weeks 5 to 8 was significantly increased (P<0.05), and the laying rate in experimental groups 1 and 2 in weeks 9 to 12 was significantly increased (P<0.05). 2) Compared with the control group, the eggshell strength in experimental groups 1 and 2 at weeks 4 and 12 and in experimental group 1 at weeks 8 was significantly increased (P<0.05); the albumen height in experimental group 1 at week 8 and in experimental group 2 at week 12 was significantly increased (P<0.05); the Haugh unit in experimental groups 1 and 2 at each stage was significantly increased (P<0.05). 3) Compared with the control group, the contents of calcium and phosphorus in serum and calcium content in tibia in experimental groups 1 and 2 were significantly increased (P<0.05). 4) Compared with the control group, the activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and catalase (CAT) as well as the superoxide anion scavenging ability and hydroxyl radical scavenging ability in serum in experimental groups 1 and 2 were significantly increased (P<0.05), and the malondialdehyde (MDA) content in serum was significantly decreased (P<0.05); the total antioxidant capacity (T-AOC) in serum in experimental group 1 was significantly increased (P<0.05). Compared with the control group, the contents of carbonic anhydrase (CA) and parathyroid hormone (PTH) in serum in experimental group 1 were significantly increased (P<0.05), and the diamine oxidase (DAO) content in serum was significantly decreased (P<0.05); the bone Gla protein (BGP) content in serum in experimental groups 1 and 2 was significantly decreased (P<0.05); the calcitonin (CT) content in serum in experimental group 2 was significantly decreased (P<0.05). 5) Compared with the control group, the villus height of jejunum in experimental group 1 was significantly increased (P<0.05), the crypt depth of jejunum in experimental group 2 was significantly decreased (P<0.05), and the villus height to crypt depth ratio of jejunum in experimental groups 1 and 2 were significantly increased (P<0.05). 6) Compared with the control group, the tibia strength, trabecular number and trabecular area in experimental group 2 were significantly increased (P<0.05), and the tibia ash content in experimental group 1 was significantly increased (P<0.05). In conclusion, compared with dietary VD3 only, dietary 25-OHD3 can improve performance and egg quality, enhance antioxidant capacity, maintain intestinal morphology, promote the absorption and deposition of calcium and phosphorus in serum and tibia, and improve tibia quality of laying hens in late laying period.

Key words： 25-hydroxyvitamin D3; laying hens in late laying period; performance; egg quality; calcium and phosphorus metabolism; tibial quality

Cite this article

GAO Shan , ZHENG Junjie , QIU Kai , ZHANG Haijun , WU Shugeng . Effects of Substituting 25-Hydroxyvitamin D3 for Vitamin D3 on Performance, Egg Quality and Calcium and Phosphorus Metabolism of Laying Hens in Late Laying Period[J]. Chinese Journal of Animal Nutrition, 2024 , 36(5) : 2992 -3002 . DOI: 10.12418/CJAN2024.257

随着蛋鸡规模化养殖的发展,低豆粕、低蛋白质饲粮的应用以及延长养殖期和无抗饲养等成为蛋鸡生产的发展趋势,改善蛋鸡生产性能和鸡蛋品质等也成为蛋鸡生产中亟待解决的问题。产蛋后期的蛋鸡因各器官老化,常伴随着生产性能(产蛋率)和蛋品质(蛋壳强度和哈氏单位)的下降,这给产蛋后期蛋鸡生产造成了严重损失。饲粮及骨骼(35%~40%)是蛋壳形成的重要钙源[1],产蛋后期机体钙和磷等吸收不足,易导致蛋鸡体内钙流失以及骨骼质量下降,表现为骨变形、骨折、骨质疏松以及笼层疲劳等[2-3];且约30%患有骨质疏松症的蛋鸡出现骨折的现象[4]。骨质量与产蛋量和蛋壳质量密切相关,与健康蛋鸡相比,患有严重骨质疏松症的蛋鸡体重下降,产蛋量下降18%,骨骼钙储备损失15%~20%[5]。因此,增加产蛋后期蛋鸡对钙的吸收和沉积,是改善其生产性能、提高蛋壳质量以及减少经济损失的重要途径。
作为脂溶性维生素,维生素D3(VD3)通过促进肠道对钙的吸收,对体内矿物质和骨骼中钙稳态起关键作用。但饲粮中长期高剂量补充VD3,会出现明显的蛋鸡体重、产蛋率及骨密度下降现象[6]。在肝脏中,VD3在羟化酶的作用下转化为25-羟基维生素D3(25-OHD3),25-OHD3进入全身循环后在肾脏经1α-羟基化酶羟化成具有活性的1,25-二羟基维生素D3[1,25-(OH)2D3][7]。25-OHD3作为VD3的活性物质,自2006年被欧盟批准作为饲料添加剂替代VD3用于家禽生产,并随着其生产成本下降,应用价值不断凸显,近年来已被广泛应用。研究表明,饲粮中添加69 μg/kg 25-OHD3 16周后,可显著降低破蛋率,提升蛋鸡胫骨质量[8]。Edelstein等[9]发现存在2种不同大小的维生素D转运蛋白质,且小蛋白质优先与25-OHD3结合。正常条件下,肠细胞中也存在优先与25-OHD3相结合的运输蛋白,相对结合亲和力为表现为25-OHD3>1,25-(OH)2D3>VD3,因此相比VD3,25-OHD3具有更高的吸收效率及生物活性[10-11]。此外,25-OHD3的生物学效价高于VD3,为1~4倍,其取决于试验中检测的指标,包括生产性能、胫骨指标、血清指标以及钙吸收代谢指标等[12]。但部分结果表明,蛋鸡饲粮中添加等量25-OHD3和VD3,2组鸡蛋比重、蛋壳质量和血浆中钙和磷含量无显著差异[13-14]。最近研究表明,与对照组相比(仅添加4 560 IU/kg VD3),饲粮中添加69 μg/kg 25-OHD3+1 800 IU/kg VD3可促进0~14周蛋鸡胫骨钙磷沉积,降低20~24周龄蛋鸡破蛋率[15],表明25-OHD3替代部分VD3的效果远高于仅添加VD3。但25-OHD3作为VD3的替代品对后期蛋鸡生产的影响尚不清楚。因此,本试验旨在研究饲粮中添加25-OHD3对55周龄海兰褐蛋鸡生产性能、蛋品质、钙磷代谢、肠道形态及胫骨质量的影响,探讨产蛋后期蛋鸡饲粮中添加25-OHD3的作用及其与VD3的替代组合关系。

1 材料与方法

1.1 试验材料

本试验所用25-OHD3产品为市售,其中25-OHD3含量为1.25%;VD3规格为5×105 IU/g。

1.2 试验设计

采用单因素完全随机设计,参考《饲料原料和饲料添加剂畜禽靶动物有效性评价试验技术指南》(GB/Z 31813—2015),选用采食正常、体重[(2.07±0.09) kg]相近的健康海兰褐蛋鸡270只(55周龄,整齐度>90%),随机分为3个组,每组6个重复,每个重复15只鸡,每笼3只鸡。对照组饲喂添加4 000 IU/kg VD3的基础饲粮,试验1组饲喂添加50 μg/kg 25-OHD3和2 000 IU/kg VD3的饲粮,试验2组饲喂添加100 μg/kg 25-OHD3的饲粮。因VD3的国际单位换算关系为1 μg/kg=40 IU/kg,所以本研究采用相等当量替换,即分别用50和100 μg/kg 25-OHD3替代基础饲粮中等量的VD3。试验设计及分组见表1。基础饲粮参照《鸡饲养标准》(NY/T 33—2004),并结合海兰褐产蛋鸡饲养手册配制,其组成及营养水平见表2。预试期1周,正试期12周。
表1 试验设计及分组

Table 1 Experimental design and grouping

项目
Items		 对照组
Control group		 试验1组
Experimental group 1		 试验2组
Experimental group 2
25-OHD3产品添加水平
25-OHD3 product supplemental levels/(g/t)		 4 8
25-OHD3含量25-OHD3 contents/% 1.25 1.25
25-OHD3添加水平
25-OHD3 supplemental levels/(μg/kg)		 50 100
VD3添加水平
VD3 supplemental levels/(IU/kg)		 4 000 2 000
表2 基础饲粮组成及营养水平(风干基础)

Table 2 Composition and nutrient levels of the basal diet (air-dry basis) %

原料Ingredients 含量Content 营养水平Nutrient levels2) 含量Content
玉米Corn 63.65 粗蛋白质CP 16.50
豆粕Soybean meal 25.41 代谢能ME/(MJ/kg) 11.30
大豆油Soybean oil 0.11 钙Ca 3.50
石粉Limestone 9.18 总磷TP 0.49
磷酸氢钙CaHPO4 0.90 有效磷AP 0.29
氯化钠NaCl 0.30 蛋氨酸+胱氨酸Met+Cys 0.65
DL-蛋氨酸DL-Met 0.18 赖氨酸Lys 0.79
预混料Premix1) 0.27 苏氨酸Thr 0.56
合计Total 100.00 色氨酸Try 0.18

1)预混料为每千克饲粮提供 The premix provided the following per kilogram of the diet:VA 12 500 IU,VE 15 IU,VK 2 mg,VB1 1 mg,VB2 8.5 mg,VB6 8 mg,VB12 5 mg,D-泛酸 D-pantothenic acid 50 mg,烟酸 nicotinic acid 32.5 mg,生物素 biotin 2 mg,叶酸 folic acid 5 mg,胆碱 choline 500 mg,Mn 65 mg,I 1 mg,Fe 60 mg,Cu 8 mg,Zn 66 mg,植酸酶 phytase 350 U。

2)营养水平中钙和总磷为测定值,分别参考GB/T 6436—2018和GB/T 6437—2018测定;其余为计算值,参考《中国饲料成分及营养价值表(2018年第29版)》计算得出。Ca and TP were measured values, which were determined according to GB/T 6436—2018 and GB/T 6437—2018, respectively; the others were calculated values based on Tables of Feed Composition and Nutrition Values in China (29th ed, 2018).

1.3 饲养管理

本试验采用密闭式鸡舍3层阶梯式笼养,将3个组的共18个重复随机分布于室内各个位置。光照制度:光照时长16 h/d,早晚补光,光照强度20 lx。室内相对湿度为50%~80%,自然通风和纵向负压通风相结合。试验鸡自由采食和饮水;每日捡蛋和自动清粪1次;每周消毒1次,试验鸡群实施常规免疫。

1.4 样品采集与指标测定

1.4.1 生产性能

正试期,每日以重复为单位记录产蛋数(包括软壳蛋、破壳蛋和沙皮蛋等畸形蛋)、日产蛋总重量以及死亡和淘汰鸡数,每2周以重复为单位记录饲粮消耗量1次。计算每周和整个试验期的产蛋率、平均日采食量、平均蛋重和料蛋比等指标。

1.4.2 蛋品质

正试期第4、8和12周末,分别以重复为单位随机选取鸡蛋5枚,采用EA-01型蛋品质自动分析仪(Egg Analyzer,Orka Food Technology Ltd.,以色列)测定蛋白高度和哈氏单位;采用蛋壳强度分析仪(Egg Force Reader,Orka Food Technology Ltd.,以色列)测定蛋壳强度;采用数显游标卡尺测定长径和短径值,并计算蛋形指数(长径值/短径值)。

1.4.3 血清激素含量和抗氧化指标

正试期第12周末,以重复为单位随机选择1只接近该重复平均体重的蛋鸡,翅静脉采血5 mL,860×g离心10 min制备血清,于-20 ℃保存待用。通过钙和磷检测试剂盒分别在660和340 nm处检测血清钙和磷含量。采用酶联免疫吸附测定(ELISA)试剂盒(上海科华生物工程股份有限公司)测定血清25-OHD3、二胺氧化酶(diamine oxidase,DAO)、碳酸酐酶(carbonic anhydrase,CA)、甲状旁腺素(parathyroid hormone,PTH)、骨钙素(bone Gla protein,BGP)和降钙素(calcitonin,CT)含量。血清超氧化物歧化酶(superoxide dismutase,SOD)、谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)、过氧化氢酶(catalase,CAT)活性以及清除超氧阴离子和羟自由基能力、丙二醛(malondialdehyde,MDA)含量、总抗氧化能力(total antioxidant capacity,T-AOC)检测使用试剂盒(南京建成生物工程研究所)测定。

1.4.4 肠道形态

将采血后的蛋鸡处死,分离空肠,取空肠中段3 cm于4%多聚甲醛溶液中固定48 h。将固定后的肠段用乙醇脱水、二甲苯透明、石蜡包埋后4 μm切片,进行苏木精-伊红(HE)染色;染色切片于40倍显微镜下观察,随机选取3个完整的典型绒毛和隐窝拍照,使用医学图像分析软件ImagePro Plus 7.0(Media Cybernetics,美国)测量绒毛高度和隐窝深度,并计算绒毛高度/隐窝深度值。

1.4.5 胫骨质量

取左侧胫骨于4%多聚甲醛溶液中固定48 h,用10%乙二胺四乙酸(EDTA)溶液脱钙,脱水后石蜡包埋,切成5 μm切片,进行甲苯胺蓝染色;切片于200倍放大扫描下进行组织学观察,随机选取3个视野拍照,使用医学图像分析软件ImagePro Plus 7.0(Media Cybernetics,美国)测量骨小梁数量及骨小梁面积。去除右侧胫骨周围的肌肉和软组织,用质构仪(TA.XT Plus,Stable Microsystems Corp.,英国)测量胫骨强度。将胫骨样本放入烘箱,于105 ℃下烘干48 h称重;经石油醚脱脂后,放入马弗炉550 ℃灰化24 h后称重,并研磨成粉末状,于室温下干燥保存。取约0.2 g粉末样本于锥形瓶,加入5 mL硝酸溶解,220 ℃电沙浴消煮3 h,超纯水定稀释625倍,过膜(0.22 μm),通过电感耦合等离子体原子发射光谱仪(Agilent Technologies Inc.,美国)测定样品中粗灰分含量。

1.5 数据统计分析

采用SPSS 20.0软件对试验数据进行单因素方差分析,差异显著者采用Turkey法进行多重比较。结果均以“平均值±标准差”表示,P<0.05表示差异显著。

2 结果与分析

2.1 饲粮中添加25-OHD3对产蛋后期蛋鸡生产性能的影响

由表3可知,饲粮中添加25-OHD3对产蛋后期蛋鸡平均蛋重、平均日采食量和料蛋比均无显著影响(P>0.05)。试验第0周(预试期),各组产蛋率无显著差异(P>0.05);与对照组相比,试验1组第5~8周产蛋率显著提高(P<0.05),试验1组和试验2组第9~12周产蛋率显著提高(P<0.05)。
表3 饲粮中添加25-OHD3对产蛋后期蛋鸡生产性能的影响

Table 3 Effects of dietary 25-OHD3 on performance of laying hens in late laying period

项目
Items		 时间
Time		 对照组
Control group		 试验1组
Experimental
group 1		 试验2组
Experimental
group 2		 均值标准误
SEM		 P值
P-value
平均蛋重
Average egg
weight/g		 第1~4周Weeks 1 to 4 60.51±2.34 60.05±2.49 59.95±1.16 0.548 0.922
第5~8周Weeks 5 to 8 61.75±3.29 59.22±1.16 59.81±1.04 0.639 0.258
第9~12周Weeks 9 to 12 61.54±1.51 61.18±2.00 59.70±1.21 0.484 0.280
平均日采食量
Average daily
feed intake/g		 第1~4周Weeks 1 to 4 122.93±1.93 122.02±3.56 121.04±1.28 0.681 0.573
第5~8周Weeks 5 to 8 124.31±3.03 117.94±5.60 124.32±3.99 2.860 0.108
第9~12周Weeks 9 to 12 125.24±13.85 118.03±9.79 124.28±5.24 2.860 0.582
料蛋比
Ratio of
feed to egg		 第1~4周Weeks 1 to 4 2.11±0.08 2.03±0.16 2.06±0.03 0.030 0.590
第5~8周Weeks 5 to 8 2.02±0.16 1.99±0.13 2.08±0.03 0.028 0.468
第9~12周Weeks 9 to 12 2.23±0.17 2.24±0.07 2.20±0.16 0.040 0.917
产蛋率
Laying rate/%		 预试期Preliminary trial period 86.67±0.07 86.67±0.08 86.78±0.04 0.015 0.999
第1~4周Weeks 1 to 4 88.83±0.70 89.36±3.16 90.06±1.60 0.566 0.715
第5~8周Weeks 5 to 8 84.20±1.99b 89.19±3.06a 88.12±1.76ab 0.913 0.027
第9~12周Weeks 9 to 12 75.39±1.38b 84.39±3.83a 81.40±3.36a 1.325 <0.001

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 superscripts mean no significant difference (P>0.05), while with different letter superscripts mean significant difference (P<0.05). The same as below.

2.2 饲粮中添加25-OHD3对产蛋后期蛋鸡蛋品质的影响

由表4可知,饲粮中添加25-OHD3对产蛋后期蛋鸡蛋形指数无显著影响(P>0.05)。与对照组相比,试验1组和试验2组第4周和第12周以及试验1组第8周蛋壳强度显著提高(P<0.05);试验1组第8周和试验2组第12周蛋白高度显著提高(P<0.05);试验1组和试验2组试验各阶段哈氏单位均显著提高(P<0.05),且试验1组和试验2组之间无显著差异(P>0.05)。
表4 饲粮中添加25-OHD3对产蛋后期蛋鸡蛋品质的影响

Table 4 Effects of dietary 25-OHD3 on egg quality of laying hens in late laying period

项目
Items		 时间
Time		 对照组
Control group		 试验1组
Experimental
group 1		 试验2组
Experimental
group 2		 均值标准误
SEM		 P值
P-value
蛋形指数
Egg shape
index		 第4周Week 4 1.24±0.01 1.25±0.00 1.24±0.01 0.002 0.091
第8周Week 8 1.34±0.02 1.33±0.00 1.34±0.00 0.004 0.539
第12周Week 12 1.30±0.12 1.29±0.02 1.32±0.02 0.005 0.119
蛋壳强度
Eggshell
strength/N		 第4周Week 4 38.10±0.65b 42.91±2.22a 41.60±0.69a 0.712 <0.001
第8周Week 8 32.49±1.25b 35.21±0.95a 33.11±1.30ab 0.466 0.024
第12周Week 12 33.06±1.07b 36.96±2.79a 39.02±1.07a 0.885 <0.001
蛋白高度
Albumen
height/mm		 第4周Week 4 6.46±0.32 7.11±0.54 7.22±0.21 0.146 0.079
第8周Week 8 6.16±0.67b 7.11±0.35a 6.99±0.10ab 0.172 0.026
第12周Week 12 6.67±0.27b 7.35±0.25ab 7.48±0.60a 0.886 0.040
哈氏单位
Haugh unit		 第4周Week 4 76.14±4.05b 82.63±3.35a 83.52±2.12a 1.141 0.021
第8周Week 8 76.52±5.09b 83.57±1.73a 82.75±0.81a 0.404 0.022
第12周Week 12 79.67±1.10b 87.58±3.40a 85.91±2.89a 1.239 <0.001

2.3 饲粮中添加25-OHD3对产蛋后期蛋鸡血清和胫骨钙和磷含量的影响

由表5可知,饲粮中添加25-OHD3对产蛋后期蛋鸡血清钙和磷含量以及胫骨钙含量有显著影响(P<0.05),对胫骨磷含量无显著影响(P>0.05)。与对照组相比,试验1组和试验2组血清钙和磷含量以及胫骨钙含量均显著提高(P<0.05);试验1组血清钙含量显著高于试验2组(P<0.05),其他指标无显著差异(P>0.05)。
表5 饲粮中添加25-OHD3对产蛋后期蛋鸡血清和胫骨钙和磷含量的影响

Table 5 Effects of dietary 25-OHD3 on contents of calcium and phosphorus in serum and tibia of laying hens in late laying period

项目
Items		 对照组
Control group		 试验1组
Experimental group 1		 试验2组
Experimental group 2		 均值标准误
SEM		 P值
P-value
血清Serum/(mol/L)
钙含量Ca content 2.36±0.07c 2.83±0.05a 2.61±0.08b 0.070 <0.001
磷含量P content 2.35±0.08b 2.66±0.06a 2.74±0.09a 0.064 <0.001
胫骨Tibia/%
钙含量Ca content 35.17±1.47b 40.15±0.15a 40.92±0.64a 0.806 <0.001
磷含量P content 17.50±0.27 17.15±0.35 17.23±0.38 0.099 0.351

2.4 饲粮中添加25-OHD3对产蛋后期蛋鸡血清抗氧化指标的影响

由表6可知,饲粮中添加25-OHD3产蛋后期蛋鸡血清抗氧化指标均有显著影响(P<0.05)。与对照组相比,试验1组和试验2组血清SOD、GSH-Px和CAT活性均显著提高(P<0.05),且试验2组血清GSH-Px和CAT活性显著高于试验1组(P<0.05);试验1组和试验2组血清清除超氧阴离子能力和清除羟自由基能力显著提高(P<0.05),血清MDA含量显著降低(P<0.05),且试验2组血清MDA含量显著低于试验1组(P<0.05);试验1组血清T-AOC显著高于对照组和试验2组(P<0.05)。
表6 饲粮中添加25-OHD3对产蛋后期蛋鸡血清抗氧化指标的影响

Table 6 Effects of dietary 25-OHD3 on serum antioxidant indices of laying hens in late laying period

项目
Items		 对照组
Control
group		 试验1组
Experimental
group 1		 试验2组
Experimental
group 2		 均值
标准误
SEM		 P值
P-value
超氧化物歧化酶SOD/(U/mL) 13.26±1.28b 16.20±1.25a 18.69±0.55a 0.843 <0.001
谷胱甘肽过氧化物酶GSH-Px/(U/mL) 515.83±18.28c 744.17±20.22b 814.72±10.35a 45.361 <0.001
过氧化氢酶CAT/(U/mL) 4.04±0.40c 11.94±0.68b 22.43±0.08a 2.667 <0.001
清除超氧阴离子能力
Superoxide anion scavenging ability/(U/mL)		 429.09±7.05b 490.89±10.49a 475.14±8.48a 9.610 <0.001
清除羟自由基能力
Hydroxyl radical scavenging ability/(U/mL)		 238.45±0.77c 250.31±1.30a 243.14±1.92b 1.771 <0.001
丙二醛MDA/(nmol/mL) 21.95±1.13a 12.56±1.02b 9.11±0.97c 1.942 <0.001
总抗氧化能力T-AOC/(U/mL) 19.19±1.13b 27.48±2.81a 21.99±0.90b 1.326 <0.001

2.5 饲粮中添加25-OHD3对产蛋后期蛋鸡血清激素含量的影响

如图1所示,与对照组相比,试验1组产蛋后期蛋鸡血清CA和PTH含量显著提高(P<0.05),血清DAO含量显著降低(P<0.05);试验1组和试验2组血清BGP含量显著降低(P<0.05),且试验2组血清BGP含量显著低于试验1组(P<0.05);试验2组血清CT含量显著降低(P<0.05)。与试验1组相比,试验2组血清25-OHD3和PTH含量显著降低(P<0.05),血清DAO含量显著提高(P<0.05)。
图1 饲粮中添加25-OHD3对产蛋后期蛋鸡血清激素含量的影响

数据柱标注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)。下图同。

Fig.1 Effects of dietary 25-OHD3 on serum hormone contents of laying hens in late laying period

Value columns with the same letter superscripts mean no significant difference (P>0.05), while with different letter superscripts mean significant difference (P<0.05). The same as below.

2.6 饲粮中添加25-OHD3对产蛋后期蛋鸡空肠形态的影响

如图2所示,饲粮中添加25-OHD3对产蛋后期蛋鸡空肠形态有显著影响(P<0.05)。与对照组相比,试验1组空肠绒毛高度显著提高(P<0.05),试验2组空肠隐窝深度显著降低(P<0.05),试验1组和试验2组空肠绒毛高度/隐窝深度值均显著提高(P<0.05)。
图2 饲粮中添加25-OHD3对产蛋后期蛋鸡空肠形态的影响

Fig.2 Effects of dietary 25-OHD3 on jejunum morphology of laying hens in late laying period (40×)

2.7 饲粮中添加25-OHD3对产蛋后期蛋鸡胫骨质量的影响

如图3所示,饲粮中添加25-OHD3后产蛋后期蛋鸡胫骨中骨小梁排列紧密,且与对照组相比,试验2组骨小梁数量和骨小梁面积显著提高(P<0.05);试验2组胫骨强度显著高于其他2组(P<0.05);试验1组胫骨粗灰分含量显著高于对照组(P<0.05)。
图3 饲粮中添加25-OHD3对产蛋后期蛋鸡胫骨质量的影响

Fig.3 Effects of dietary 25-OHD3 on tibia quality of laying hens in late laying period (200×)

3 讨论

与VD3相比,25-OHD3作为家禽饲料添加剂,用于改善蛋鸡生产性能、蛋壳质量和维持骨骼健康的研究结果并不一致。产蛋后期蛋鸡生物转化系统老化,转化效率降低,VD3转化成25-OHD3(VD3的主要循环形式)的效率降低,理论上同等效价25-OHD3比VD3更能促进钙在骨骼和蛋壳形成过程中的吸收[16]。然而一项研究表明,在不同钙水平下(3.34%、4.30%、4.73%和4.94%),饲粮中添加2 760 IU/kg VD3(相当于69 μg/kg VD3)或69 μg/kg 25-OHD3 16周,对蛋鸡生产性能和蛋壳品质无显著影响[13]。维生素D在禽类肠道和骨骼钙和磷的代谢中起着重要作用[17],蛋鸡饲粮中长期(0~95周)添加69 μg/kg 25-OHD3+2 760 IU/kg VD3,与仅添加VD3相比,可提高血清25-OHD3含量,改善骨结构质量,促进骨皮质区的矿物质沉积,增加骨小梁的吸收,为产蛋后期蛋鸡矿物质沉积提供更多的空间[18]。本研究表明,与对照组相比,饲喂添加50 μg/kg 25-OHD3+2 000 IU/kg VD3饲粮8周(试验1组),产蛋率显著提高,且试验1组第9~12周产蛋率提高了11.94%;此外,蛋壳强度、蛋白高度和哈氏单位在仅添加25-OHD3 4周后具有显著改善。此结果与之前的结果一致,饲粮中添加25-OHD3提高蛋鸡低放养密度下产蛋率,降低破蛋率[19]。这表明饲粮中添加25-OHD3能够提高产蛋后期蛋鸡生产性能和蛋品质,且增强蛋壳质量的效果优于仅添加VD3。
研究表明,具有活性的代谢产物25-OHD3比VD3能更有效地提高血液中25-OHD3含量,且在肠道中消化吸收得更快[20-21],血清25-OHD3含量是评价体内VD3状态的重要指标。正常条件下,体内钙含量的稳态是通过肠道吸收、肾脏排泄和骨代谢来实现,这些过程由PTH、CT、VD3含量和性激素共同调节[22]。CA可从过氧化氢(H2O2)和二氧化碳(CO2)中产生质子和碳酸根离子(C O 3 2 -),是参与蛋壳形成过程中碳酸钙沉积的关键酶,并参与骨骼健康和酸碱调节[23]。研究表明,血清中较高含量的25-OHD3有利于蛋鸡维持钙和骨骼稳态[19]。本试验中,与对照组相比,饲粮中添加50 μg/kg 25-OHD3+2 000 IU/kg VD3可提高产蛋后期蛋鸡血清25-OHD3、CA和PTH含量;饲粮中添加100 μg/kg 25-OHD3可降低血清BGP和CT含量,提高血清和胫骨钙含量,这与之前的研究结果一致。由此说明,25-OHD3可通过调节激素分泌,促进血液中的钙吸收及胫骨中钙沉积。
在蛋鸡中,钙的吸收主要发生在小肠,肠道中存在VD3诱导的钙结合蛋白,在蛋壳形成期间,能够促进十二指肠和空肠对钙的吸收[24]。研究表明,饲粮中添加12.5 μg/kg 25-OHD3可提高肉仔鸡小肠中维生素D结合受体及磷转运蛋白的表达[25]。通过与胚蛋注射VD3相比,胚蛋注射25-OHD3可提高肉仔鸡十二指肠、空肠和回肠绒毛高度/隐窝深度值,改善小肠形态,促进营养吸收[26]。随着饲养周期延长,蛋鸡血清和肝脏中抗氧化酶(SOD和GSH-Px)活性下降,导致产蛋后期蛋鸡抗氧化能力降低,空肠绒毛高度显著降低,隐窝深度明显提高,这阻碍了营养物质的吸收,损伤了蛋鸡肠道健康[27-28]。DAO含量是评价肠道损伤的重要标志物之一,空肠屏障结构和功能的破坏导致DAO含量增加[29]。研究表明,与相同水平的VD3相比,饲粮中添加25-OHD3显著提升肠道抗氧化酶活性,降低MDA含量,并维持肠道完整性[30]。本试验中,饲粮中添加25-OHD3能够提高产蛋后期蛋鸡血清抗氧化酶SOD、GSH-Px和CAT活性,增强清除超氧阴离子和羟自由基能力,并降低血清MDA和DAO含量,改善空肠形态。因此,25-OHD3可通过调节产蛋后期蛋鸡抗氧化防御系统,改善肠道形态,促进钙的吸收和沉积。
钙磷利用差和骨骼矿化不足是导致蛋鸡骨骼问题的主要原因,饲粮中添加VD3是改善骨骼结构的途径之一。VD3通过调节小肠对钙和磷的吸收,保持血清钙和磷在骨矿化和骨重塑中的动态平衡[31]。研究表明,维生素D影响骨骼肌的代谢和功能,VD3及其代谢物(25-OHD3)可提高机体对钙和磷的吸收,增强胫骨矿化和断裂强度,增加骨密度[32⇓-34]。本试验中,饲粮中添加100 μg/kg 25-OHD3显著提高产蛋后期蛋鸡胫骨强度以及骨小梁数量和面积,饲粮中添加50 μg/kg 25-OHD3+2 000 IU/kg VD3显著提高胫骨粗灰分含量。这与前人研究结果一致,饲粮中添加25-OHD3使蛋鸡胫骨骨小梁连接紧密,并能够降低骨折现象,改善胫骨质量[19]。由此可见,与饲粮中仅添加VD3相比,饲粮中添加25-OHD3可改善产蛋后期蛋鸡胫骨质量。

4 结论

与饲粮中仅添加VD3相比,饲粮中添加25-OHD3可提高产蛋后期蛋鸡生产性能和蛋品质,增强抗氧化能力,维持肠道形态,促进血清和胫骨钙和磷的吸收和沉积,改善胫骨质量。

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