Chinese Journal of Animal Nutrition >

2026 , Vol. 38 >Issue 7: 5078 - 5090

DOI: https://doi.org/10.12418/CJAN2026.407

RESEARCH PAPER

Effects of Dietary Calcium and Non-Phytate Phosphorus Levels on Laying Performance, Digestive Function and Egg Quality of Langya Hens During Early Laying Period

  • ZHU Yimin , 1 ,
  • SHI Xueping 1 ,
  • ZHOU Chuanfeng 2 ,
  • ZHANG Bolin 1 ,
  • GONG Xiangwei 3 ,
  • WANG Shubai , 1, *
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  • 1 College of Animal Science and Technology, Qingdao Agricultural University, Qingdao 266109, China
  • 2 College of Life Sciences, Qingdao Agricultural University, Qingdao 266109, China
  • 3 Shandong Qingdao Langya Chicken Breeding Co., Ltd., Qingdao 266500, China
* professor, E-mail:

Received date: 2025-12-25

  Online published: 2026-07-14

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Abstract

This experiment was conducted to investigate the effects of dietary calcium (Ca) and non-phytate phosphorus (NPP) levels on laying performance, egg quality, intestinal digestive enzyme activities, nutrient apparent utilization rates and gene expression of osteopontin (OPN) and calbindin (CALB) in eggshell gland of Langya hens during early laying period. A total of 540 healthy 168-day-old Langya chickens were selected and randomly divided into 9 groups according to the principle of equal body weight and laying rate, with 5 replicates per group and 12 chickens per replicate. A 2×3 two-factor orthogonal experimental design was adopted, the dietary Ca levels were set at 2.50%, 3.00% and 3.50%, respectively, and the NPP levels were set at 0.24%, 0.32% and 0.40%, respectively. The experiment included a 7-day pre-trial period and a 35-day formal trial period. The results showed as follows: 1) the average daily egg mass and laying rate of 2.50% and 3.00% Ca groups were significantly higher than those of 3.50% Ca group (P<0.05), and the feed to egg ratio was significantly lower than that of the 3.50% Ca group (P<0.05); dietary Ca and NPP levels had significant interaction effects on the average daily egg mass, laying rate and feed to egg ratio (P<0.05). 2) The Haugh unit of 3.00% Ca group was significantly higher than that of 2.50% and 3.50% Ca groups (P<0.05), and the eggshell strength of 3.00% and 3.50% Ca groups was significantly higher than that of 2.50% Ca group (P<0.05); the eggshell strength of 0.40% NPP group was significantly higher than that of 0.24% and 0.32% NPP groups (P<0.05); dietary Ca and NPP levels had significant interaction effects on albumen height, Haugh unit and eggshell strength (P<0.05). 3) The intestinal trypsin activity of 3.00% Ca group was significantly higher than that of 2.50% and 3.50% Ca groups (P<0.05); the intestinal trypsin activity of 0.32% NPP group was significantly higher than that of the 0.24% and 0.40% NPP groups (P<0.05); dietary Ca and NPP levels had a significant interaction effect on intestinal trypsin activity (P<0.05). 4) The apparent utilization rates of crude protein and Ca of 3.00% Ca group were significantly higher than those of 3.50% Ca group (P<0.05), and the apparent utilization rate of phosphorus of 3.00% Ca group was significantly higher than that of 2.50% and 3.50% Ca groups (P<0.05); the apparent utilization rate of crude protein of 0.32% NPP group was significantly higher than that of 0.24% and 0.40% NPP groups (P<0.05); dietary Ca and NPP levels had significant interaction effects on the apparent utilization rates of Ca and phosphorus (P<0.05). 5) The mRNA relative expression level of CALB in eggshell gland of 3.00% and 3.50% Ca groups was significantly higher than that of 2.50% Ca group (P<0.05); the mRNA relative expression levels of OPN and CALB in eggshell gland of 0.32% and 0.40% NPP groups were significantly higher than those of 0.24% NPP group (P<0.05); dietary Ca and NPP levels had significant interaction effects on the mRNA relative expression levels of OPN and CALB in eggshell gland (P<0.05). In conclusion, the dietary appropriate Ca and NPP levels can improve the laying performance and egg quality of Langya hens during early laying period, enhance the digestive function, and increase the mRNA relative expression levels of OPN and CALB in eggshell gland. The optimal dietary Ca and NPP levels for Langya hens during early laying period are 3.00% and 0.32%, respectively.

Key words： Langya hens; calcium; non-phytate phosphorus; laying performance; egg quality; digestive function; osteopontin; calbindin

Cite this article

ZHU Yimin , SHI Xueping , ZHOU Chuanfeng , ZHANG Bolin , GONG Xiangwei , WANG Shubai . Effects of Dietary Calcium and Non-Phytate Phosphorus Levels on Laying Performance, Digestive Function and Egg Quality of Langya Hens During Early Laying Period[J]. Chinese Journal of Animal Nutrition, 2026 , 38(7) : 5078 -5090 . DOI: 10.12418/CJAN2026.407

琅琊鸡原产于山东省日照市、胶南市、诸城市等地,由于这些区域在历史行政区划中隶属于琅琊郡,琅琊鸡这一名称也由此而来[1]。20世纪70年代,琅琊鸡凭借其独特的品种优势被列为山东地方家禽良种;随着国家对畜禽遗传资源保护力度的加大,琅琊鸡于2008年入选《国家畜禽遗传资源品种名录》,于2009年入选《山东省畜禽遗传资源保护名录》,是我国备受关注的优良地方家禽品种[2]。琅琊鸡具有体型小、抗病力强、适应性好、蛋品质高等优点[3]。研究发现,琅琊鸡鸡蛋的蛋黄指数和卵磷脂含量均显著高于海兰褐蛋鸡[4]。但是,琅琊鸡存在产蛋率低、饲料报酬低、养殖成本较高等缺点。目前,我国缺少针对琅琊鸡饲粮营养需要量的系统性研究,生产中琅琊鸡的饲粮配制,其营养水平多参考我国现行的《黄羽肉鸡营养需要量》(NY/T 3645—2020)、蛋鸡饲养标准及养殖户经验等,因此,可能存在其营养供给不适宜的问题。
钙(Ca)和磷(P)是蛋鸡高产稳产的重要矿物元素[5-6]。Ca是蛋鸡骨骼生长与沉积的主要成分,同时也是蛋壳形成的核心原料,直接影响蛋壳的强度、厚度及完整性;P则参与蛋鸡体内能量代谢、核酸合成及骨骼发育等多个生理过程,对维持机体正常生理功能至关重要[7]。目前,琅琊鸡产蛋期饲粮中Ca和非植酸磷(NPP)的适宜水平研究缺乏。因此,本试验旨在探究饲粮Ca和NPP水平对产蛋前期琅琊鸡产蛋性能、蛋品质、消化机能以及蛋壳腺矿化相关基因和Ca转运相关基因表达的影响,明确产蛋前期琅琊鸡饲粮Ca和NPP的适宜水平,从而为其饲粮的合理配制提供科学依据。

1 材料与方法

1.1 伦理声明

动物试验由青岛农业大学动物科技学院实验动物伦理委员会审查批准,伦理批准编号为DKY20240522。

1.2 试验设计和饲养管理

选取168日龄健康琅琊鸡540只,根据体重和产蛋率均等原则随机分为9个组,每组5个重复,每个重复12只鸡。采用3×3二因素三水平试验设计,饲粮Ca和NPP水平参考《黄羽肉鸡营养需要量》(NY/T 3645—2020)及我国其他地方品种鸡的研究结果[8-9],Ca水平分别设置为2.50%、3.00%、3.50%,NPP水平分别设置为0.24%、0.32%、0.40%。试验分组见表1。预试期7 d,正试期35 d。
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
试验饲粮组成及营养水平见表2,除Ca、总磷(TP)和NPP外,其他营养水平基本一致。饲粮粗蛋白质、Ca、TP、氨基酸和NPP的含量分别采用GB/T 6432—2018[10]、GB/T 6436—2018[11]、GB/T 6437—2018[12]、GB/T 18246—2019[13]、李培培等[14]的方法检测,代谢能参照《中国饲料成分及营养价值表(2023年第34版)》进行计算。试验鸡均采用3层笼养,饲养于同栋鸡舍,自由采食和饮水,采用智能化环境控制设备调控舍内温度、相对湿度等环境条件。

1.3 检测指标与方法

1.3.1 产蛋性能

试验期间以重复为单位,记录采食量、产蛋总数、日产蛋总重量、破软壳蛋数和蛋鸡死淘数等数据,计算全期平均日产蛋量、平均蛋重、产蛋率、料蛋比和平均日采食量。

1.3.2 蛋品质指标

于正试期第35天,每组每个重复随机取2枚新鲜蛋进行蛋品质测定。蛋形指数和蛋壳厚度参照杨宁[15]的方法测定,蛋壳强度采用蛋壳强度测定仪(MODEL-1,日本Robotmation公司)测定,蛋重、蛋黄颜色、蛋白高度、哈氏单位采用蛋品质自动分析仪(EMT-7300,日本Robotmation公司)测定。
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

于正试期第35天,每组每个重复随机取2只蛋鸡,颈部放血致死,取十二指肠食糜,装于2 mL冻存管中,-80 ℃保存待测。采用酶标仪(SYNERGY H4,美国伯腾仪器有限公司)测定α-淀粉酶和脂肪酶活性,采用紫外可见分光光度计(UV755B,上海佑科仪器仪表有限公司)测定胰蛋白酶活性。以上指标均采用试剂盒测定,试剂盒购自南京建成生物工程研究所。

1.3.4 养分表观利用率

采用内源指示剂法进行代谢试验,以盐酸不溶灰分作为内源指示剂,于正试期第32天开始,笼下放粪盘,连续3 d收集每个重复的新鲜鸡粪,捡干净粪便中沾有的羽毛,并用10%盐酸固氮(每100 g鲜粪加入10 mL盐酸),粪样于-20 ℃保存待测。粪样和饲粮样品于65 ℃烘干制备风干样,粉碎过40目筛,粗蛋白质、粗灰分、粗脂肪、Ca、P和盐酸不溶灰分含量分别采用GB/T 6432—2018[10]、GB/T 6438—2007[16]、GB/T 6433—2006[17]、GB/T 6436—2018[11]、GB/T 6437—2018[12]、GB/T 23742—2009[18]的方法测定。养分表观利用率计算公式如下:
养分表观利用率(%)=[(饲粮中该养分含量-粪便中该养分含量)/饲粮中该养分含量]×100。

1.3.5 蛋壳腺矿化相关基因和Ca转运相关基因表达

于正试期第35天,每组每个重复随机取2只蛋鸡,颈部放血致死,取出蛋壳腺,用无菌载玻片刮取内层组织于2 mL冻存管中,于-80 ℃保存待测。检测蛋壳腺矿化相关基因骨桥蛋白(osteopontin,OPN)和Ca转运相关基因钙结合蛋白(calbindin,CALB)的mRNA相对表达量[19]。按照RNA试剂盒(货号AG21102,艾科瑞生物工程有限公司)说明书提取蛋鸡蛋壳腺组织中的总RNA,采用反转录试剂盒(货号AG11728,艾科瑞生物工程有限公司)进行反转录,采用荧光定量试剂盒(货号AG11701,艾科瑞生物工程有限公司)进行相关基因表达量的测定。基因引物序列见表3。以β-肌动蛋白(β-actin)为内参基因,采用2-ΔΔCt方法计算目的基因的mRNA相对表达量。
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

采用SPSS 26.0软件对试验数据进行双因素方差分析,饲粮Ca、NPP水平的主效应以及两者间的互作效应分析;当交互作用显著时,对组间进行单因素方差分析,并采用Turkey比较法进行显著性检验。P<0.05表示差异显著,P>0.05表示差异不显著,结果以平均值和均值标准误(SEM)表示。

2 结果

2.1 饲粮Ca和NPP水平对琅琊鸡产蛋性能的影响

由表4可知,主效应分析显示,饲粮Ca水平对平均日产蛋量、产蛋率、料蛋比有显著影响(P<0.05),2.50%和3.00% Ca组平均日产蛋量和产蛋率显著高于3.50% Ca组(P<0.05),2.50%和3.00% Ca组料蛋比显著低于3.50% Ca组(P<0.05)。互作效应分析显示,饲粮Ca和NPP水平对平均日产蛋量、产蛋率、料蛋比有显著互作效应(P<0.05)。1组、5组平均日产蛋量和产蛋率显著高于其他各组(P<0.05),1组、2组、5组料蛋比显著低于4组、8组(P<0.05)。
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

由表5可知,主效应分析显示,饲粮Ca水平对哈氏单位和蛋壳强度有显著影响(P<0.05),3.00% Ca组哈氏单位显著高于2.50%和3.50% Ca组(P<0.05),3.00%和3.50% Ca组蛋壳强度显著高于2.50% Ca组(P<0.05);饲粮NPP水平对蛋壳强度有显著影响(P<0.05),0.40% NPP组蛋壳强度显著高于0.24%和0.32% NPP组(P<0.05)。互作效应分析显示,饲粮Ca和NPP水平对蛋白高度、哈氏单位和蛋壳强度有显著互作效应(P<0.05)。6组蛋白高度显著高于5组、9组(P<0.05),6组哈氏单位显著高于1组、2组、5组、7组、9组(P<0.05),9组蛋壳强度显著高于1组、2组、4组、7组(P<0.05)。
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

由表6可知,主效应分析显示,饲粮Ca水平对肠道胰蛋白酶活性有显著影响(P<0.05),3.00% Ca组肠道胰蛋白酶活性显著高于2.50%和3.50% Ca组(P<0.05);饲粮NPP水平对肠道胰蛋白酶活性有显著影响(P<0.05),0.32% NPP组肠道胰蛋白酶活性显著高于0.24%和0.40% NPP组(P<0.05)。互作效应分析显示,饲粮Ca和NPP水平对肠道胰蛋白酶活性有显著互作效应(P<0.05)。5组肠道胰蛋白酶活性显著高于其他各组(P<0.05)。
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

由表7可知,主效应分析显示,饲粮Ca水平对粗蛋白质、Ca和P表观利用率有显著影响(P<0.05),3.00% Ca组粗蛋白质和Ca表观利用率显著高于3.50% Ca组(P<0.05),3.00% Ca组P表观利用率显著高于2.50%和3.50% Ca组(P<0.05);饲粮NPP水平对粗蛋白质表观利用率有显著影响(P<0.05),0.32% NPP组粗蛋白质表观利用率显著高于0.24%和0.40% NPP组(P<0.05)。互作效应分析显示,饲粮Ca和NPP水平的对Ca和P表观利用率有显著互作效应(P<0.05)。5组Ca和P表观利用率显著高于1组、3组、4组、8组、9组(P<0.05)。
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

由表8可知,主效应分析显示,饲粮Ca水平对蛋壳腺CALB mRNA表达量有显著影响(P<0.05),3.00%和3.50% Ca组蛋壳腺CALB mRNA相对表达量显著高于2.50% Ca组(P<0.05);饲粮NPP水平对蛋壳腺OPN和CALB mRNA相对表达量有显著影响(P<0.05),0.32%和0.40% NPP组蛋壳腺OPN和CALB mRNA相对表达量显著高于0.24% NPP组(P<0.05)。互作效应分析显示,饲粮Ca和NPP水平对蛋壳腺OPN和CALB mRNA相对表达量有显著互作效应(P<0.05)。5组、6组、8组、9组蛋壳腺OPN mRNA相对表达量显著高于1组、4组、7组(P<0.05),5组、8组、9组蛋壳腺CALB mRNA相对表达量显著高于1组、2组、3组(P<0.05)。
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

3 讨论

3.1 饲粮Ca和NPP水平对产蛋前期琅琊鸡产蛋性能的影响

Härtel[20]研究表明,产蛋前期白来航鸡饲喂不同Ca和TP水平的饲粮,3.00% Ca+0.72% TP和3.00% Ca+0.82% TP组获得较高产蛋量;2.50% Ca组平均日采食量显著高于其他各组,1.62% TP组平均日采食量显著高于其他各组;饲粮Ca和TP水平对产蛋重和平均日采食量有显著互作效应。高树华[21]研究表明,产蛋高峰期海兰褐蛋鸡饲喂不同Ca和NPP水平的饲粮,3.00% Ca+0.28% NPP组、3.50% Ca+0.32% NPP组和4.00% Ca+0.32% NPP组的料蛋比显著低于4.00% Ca+0.28% NPP组,饲粮Ca和NPP水平对产蛋率、平均蛋重、平均日产蛋量和料蛋比无显著互作效应。本试验结果表明,主效应分析显示,2.50%和3.00% Ca组琅琊鸡的平均日产蛋量和产蛋率显著高于3.50% Ca组,料蛋比则反之;饲粮NPP水平对产蛋性能无显著影响;饲粮Ca和NPP水平对平均日产蛋量、产蛋率、料蛋比有显著互作效应。据报道,饲粮Ca、P水平不适宜不仅会形成不溶且不可吸收的Ca-植酸螯合物[22],还会抑制P的吸收[23]。由此可见,饲粮Ca、P水平不适宜不仅会形成不溶且不可吸收的Ca-植酸螯合物,降低对养分的吸收,还会抑制P的吸收,进而导致产蛋性能下降。

3.2 饲粮Ca和NPP水平对产蛋前期琅琊鸡蛋品质的影响

蛋白高度、蛋黄颜色、哈氏单位、蛋壳强度、蛋壳厚度等是常用的蛋品质衡量指标[24]。张霞等[6]研究表明,22~40周龄京红1号蛋鸡饲喂不同Ca水平的饲粮,3.91% Ca组蛋壳强度显著高于3.32%和3.59% Ca组,蛋壳厚度各组间无显著差异。高树华[21]研究表明,产蛋高峰期海兰褐蛋鸡饲喂不同Ca和NPP水平的饲粮,3.50% Ca+0.32% NPP组哈氏单位最高,显著高于3.50% Ca+0.36% NPP组和4.00% Ca+0.32% NPP组,3.00% Ca+0.32% NPP组、3.50% Ca+0.36% NPP组和4.00% Ca+0.28% NPP组蛋壳厚度显著高于3.50% Ca+0.28% NPP组、3.50% Ca+0.32% NPP组和4.00% Ca+0.36% NPP组;主效应分析表明,3.00%和4.00% Ca组蛋壳厚度显著高于3.50% Ca组;饲粮Ca和NPP水平对哈氏单位和蛋壳厚度均有显著互作效应,对蛋形指数、蛋壳强度、蛋白高度和蛋黄颜色无显著互作效应。本试验结果表明,主效应分析显示,3.00% Ca组哈氏单位显著高于2.50%和3.50% Ca组,3.00%和3.50% Ca组蛋壳强度显著高于2.50% Ca组,0.40% NPP组蛋壳强度显著高于0.24%和0.32% NPP组;3.00% Ca+0.40% NPP组的蛋白高度和哈氏单位最高,3.50% Ca+0.40% NPP组的蛋壳强度最高;饲粮Ca和NPP水平对蛋白高度、哈氏单位和蛋壳强度均有显著互作效应,对蛋重、蛋黄颜色、蛋形指数和蛋壳厚度无显著互作效应。据报道,适宜的Ca水平能够促进鸡蛋形成过程中蛋白质的分泌,增强蛋白质稠度[25]。李春[26]研究报道,Ca和NPP的均衡供给可使碳酸钙晶体在蛋壳充分沉积,显著提升蛋壳强度。由此可见,饲粮Ca、P水平通过影响输卵管蛋白质分泌和碳酸钙在蛋壳上的沉积,从而影响蛋品质。目前有关Ca、P水平对不同品种蛋鸡蛋品质的影响机制研究报道较少,有待进一步研究。

3.3 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

邱梅平等[27]研究表明,18~42周龄伊莎褐蛋鸡饲喂不同NPP水平(0.29%、0.41%)饲粮,对肠道脂肪酶、胰蛋白酶和淀粉酶活性没有显著影响。本试验结果表明,主效应分析显示,3.00% Ca组肠道胰蛋白酶活性显著高于2.50%和3.50% Ca组,0.32% NPP组肠道胰蛋白酶活性显著高于0.24%和0.40% NPP组,饲粮Ca和NPP水平对肠道胰蛋白酶活性有显著互作效应。由此可见,饲粮Ca、P水平对产蛋前期琅琊鸡肠道消化酶活性有不同程度的影响,饲粮Ca水平3.00%、NPP水平0.32%利于提高消化酶活性。据报道,Ca水平过高会抑制蛋鸡胰蛋白酶活性[28],适宜的Ca水平能够维持人肠激酶活性,进而激活胰蛋白酶原,提高胰蛋白酶活性[29];适宜的P水平有助于维持较高的胰蛋白酶活性,从而促进养分的消化和吸收[30]。

3.4 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

高树华[21]研究表明,产蛋高峰期海兰褐蛋鸡饲喂不同Ca和NPP水平的饲粮,主效应分析显示,3.00%和3.50% Ca组粗蛋白质表观利用率显著高于4.00% Ca组,4.00% Ca组P表观利用率显著高于3.00% Ca组;0.32%和0.36% NPP组粗蛋白质表观利用率显著高于0.28% NPP组,0.32% NPP组P表观利用率显著高于0.28% NPP组;饲粮Ca和NPP水平对粗蛋白质、Ca、P表观利用率有显著互作效应。本试验结果表明,主效应分析显示,3.00% Ca组粗蛋白质、Ca和P表观利用率显著高于3.50% Ca组,0.32% NPP组粗蛋白质表观利用率显著高于0.24%和0.40% NPP组;饲粮Ca和NPP水平对Ca和P表观利用率有显著互作效应。据报道,适宜的饲粮P水平可改善肉鸭肠道形态结构,从而提高对养分的消化吸收能力[31]。孙利娜等[32]报道,Ca、P水平失衡易形成不溶性磷酸氢钙,从而降低蛋鸡肠道对饲粮养分的消化吸收。由此可见,适宜的饲粮Ca、P水平通过改善家禽的肠道结构进而促进肠道对养分的消化和吸收。

3.5 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

Shet等[19]研究报道,不同NPP水平饲粮中添加250、500 FTU/kg植酸酶,与0.33% NPP组相比,0.16%、0.24% NPP组23周龄白来航蛋鸡蛋壳腺OPN mRNA相对表达量显著上调,CALB mRNA相对表达量显著下调。Li等[33]研究表明,正常P水平(0.50%)、低Ca水平(0.60%)饲粮及正常Ca水平(1.10%)、低P水平(0.27%)饲粮均可使爱拔益加肉鸡肠道CALB mRNA相对表达量显著上调。本试验结果表明,主效应分析显示,3.00%和3.50% Ca组蛋壳腺CALB mRNA相对表达量显著高于2.50% Ca组,0.32%和0.40% NPP组蛋壳腺OPN和CALB mRNA相对表达量显著高于0.24% NPP组,饲粮Ca和NPP水平对蛋壳腺OPN和CALB mRNA相对表达量有显著互作效应。据报道,高Ca饲粮能够使蛋壳腺CALB mRNA相对表达量增加,促进Ca在蛋壳中的沉积,从而提高蛋壳质量[34]。Shet等[19]研究报道,23周龄白来航蛋鸡蛋壳腺中生物可利用P含量的增加能够触发OPN mRNA的表达,Ca、P比失衡能够影响肠道Ca通量,致使蛋壳腺CALB mRNA相对表达量下调。OPN是输卵管蛋壳腺中的重要蛋白[35],是参与钙离子(Ca2+)吸收的主要蛋白,其含量影响蛋壳腺内Ca2+转运和蛋壳形成[36]。OPN可以通过晶体定向和钙化速度调节蛋壳栅栏层生长,改变蛋壳晶体表面断裂的抵抗力[37]。CALB是子宫管状腺细胞Ca2+转运分泌的主要蛋白[38]。蛋壳腺中CALB以昼夜节律的方式合成[39],被称为Ca扩散的促进剂[33]。由此可见,适宜的饲粮Ca和NPP水平有助于提高鸡蛋壳腺CALB和OPN mRNA相对表达量,从而促进蛋壳中Ca沉积,提高蛋壳质量。

4 结论

饲粮Ca水平为3.00%、NPP水平为0.32%可提高产蛋前期琅琊鸡肠道消化酶活性,从而提高养分表观利用率,使产蛋性能达到最佳;通过上调蛋壳腺OPN和CALB mRNA相对表达量,使蛋壳质量达到最佳。因此,产蛋前期琅琊鸡饲粮Ca和NPP水平分别以3.00%和0.32%为宜。

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