Chinese Journal of Animal Nutrition >

2025 , Vol. 37 >Issue 11: 7455 - 7466

DOI: https://doi.org/10.12418/CJAN2025.607

RESEARCH PAPER

Effects of Dietary Isoleucine Level on Growth Performance, Meat Quality, Amino Acid Transport and Lipid Metabolism of Medium-Growing Yellow-Feathered Chickens

  • FAN Qiuli , 1 ,
  • GOU Zhongyong 1 ,
  • ZHANG Sai 1 ,
  • WANG Yibing 1 ,
  • LIN Xiajing 1 ,
  • LUO Qili 1 ,
  • DONG Xiaoli 2 ,
  • DENG Yuanfan 2 ,
  • RUAN Dong , 1, * ,
  • JIANG Shouqun , 1, *
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  • 1 Guangdong Provincial Key Laboratory of Animal Breeding and Nutrition, Key Laboratory of Animal Nutrition and Feed Science in South China, Ministry of Agriculture and Rural Affairs, State Key Laboratory of Livestock and Poultry Breeding, Institute of Animal Science, Guangdong Academy of Agricultural Sciences, Guangzhou 510640, China
  • 2 CJ (Shanghai) International Trading Co., Ltd., Shanghai 201107, China
*RUAN Dong, professor, E-mail: ;
JIANG Shouqun, professor, E-mail:

Received date: 2025-03-31

  Online published: 2025-11-14

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Abstract

This experiment was aimed to study the effects of dietary isoleucine level on growth performance, plasma biochemical indices, carcass traits, meat quality, jejunal amino acid transport and liver lipid metabolism related gene expression of medium-growing yellow-feathered chickens aged 61 to 80 days, and determined the optimal dietary isoleucine level at this stage. A total of one thousand and eighty 61-day-old female partridge chickens with an initial body weight of 1 581.67 g were randomly allocated to 6 groups with dietary isoleucine levels of 0.46%, 0.54%, 0.62%, 0.70%, 0.78% and 0.86%, respectively, and each of which included 6 replicates of 30 chickens per replicate. The experiment lasted for 20 days. The results showed as follows, with the increasing of dietary isoleucine levels: 1) the ratio of feed to gain of yellow-feathered chickens had linear and quadratic changes (P<0.05); 2) the glucose content in plasma of yellow-feathered chickens was significantly increased (P<0.05), the triglyceride content in plasma was significantly decreased (P<0.05), and the activities of alanine aminotransferase and creatine kinase had linear and quadratic changes (P<0.05); 3) the abdominal fat rate of yellow-feathered chickens was linearly decreased (P<0.05); 4) the breast muscle brightness (L*) value of yellow-feathered chickens was significantly decreased at 24 h after slaughter (P<0.05), the breast muscle redness (a*) value had linear and quadratic changes at 24 h after slaughter (P<0.05), and the shear force of breast muscle was linearly decreased (P<0.05); 5) the excitatory amino acid transporter 3 (EAAT3) gene expression in jejunum of yellow-feathered chickens had linear and quadratic changes (P<0.05); 6) the 3-hydroxy-3-methylglutaryl-CoA (HMGCR) gene expression in liver of yellow-feathered chickens was significantly decreased (P<0.05), meanwhile, the silence information regulatory factor 1 (SIRT1) gene expression in liver was significantly increased (P<0.05). In conclusion, an appropriate dietary level of isoleucine can improve the growth performance, carcass traits and meat quality of medium-growing female yellow-feathered chickens aged 61 to 80 days, while regulating plasma glucose and lipid contents as well as the expression of jejunal excitatory amino acid transporter gene and liver lipid metabolism related genes. According to the estimation of the quadratic regression model, the dietary isoleucine level for medium-growing female yellow-feathered chickens aged 61 to 80 days to achieve the best growth performance, carcass traits and meat quality is 0.68% to 0.90%. Among them, taking the ratio of feed to gain as the main evaluation index, and the optimal dietary isoleucine level is 0.74%, with a daily requirement of 742 mg.

Key words： isoleucine; yellow-feathered chickens; growth performance; plasma biochemical indices; meat quality; amino acid transport; lipid metabolism

Cite this article

FAN Qiuli , GOU Zhongyong , ZHANG Sai , WANG Yibing , LIN Xiajing , LUO Qili , DONG Xiaoli , DENG Yuanfan , RUAN Dong , JIANG Shouqun . Effects of Dietary Isoleucine Level on Growth Performance, Meat Quality, Amino Acid Transport and Lipid Metabolism of Medium-Growing Yellow-Feathered Chickens[J]. Chinese Journal of Animal Nutrition, 2025 , 37(11) : 7455 -7466 . DOI: 10.12418/CJAN2025.607

异亮氨酸作为主要支链氨基酸,除具有参与合成激素和酶类等前体功能外[1],还在机体能量稳态[2]、营养代谢[3-4]、免疫[5]和肠道健康调节[6]等方面发挥着重要作用;同时,异亮氨酸又是机体内唯一兼具生糖和生酮作用的支链氨基酸,其过量或缺乏均可造成机体免疫功能和营养物质代谢障碍[7-8]。研究表明,异亮氨酸在鸡上的应用主要体现在对生长性能、肉品质、蛋品质和肠道健康等方面的影响:异亮氨酸水平显著影响快大型黄羽肉鸡不同生长阶段平均日增重、平均日采食量和料重比,且存在性别效应[9];0.35%~0.95%异亮氨酸显著影响黄羽肉种鸡采食量、产蛋率和蛋重[10];肉鸡饲粮中添加400和800 mg/kg异亮氨酸不仅能提高胸肌和腿肌肉色亮度(L*)值和红度(a*)值,还可提高腿肌中粗蛋白质含量[11];0.72%异亮氨酸对产蛋高峰期蛋鸡产蛋率、平均蛋重、蛋白高度、蛋黄比重和蛋壳厚度均有改善作用[12];饲粮中添加2%和5%异亮氨酸可同时提高肉鸡空肠和回肠绒毛长度[13]。本课题组前期研究发现,在相同营养条件下,中速型黄羽肉鸡肌肉中风味氨基酸和脂肪酸组成优于慢速型和快速型黄羽肉鸡[14],说明中速型黄羽肉鸡在为消费者提供优质营养方面具有一定优势。加之近年来,饲用豆粕减量替代政策持续推进,而豆粕减量替代则以精准营养为基础,同时,异亮氨酸是肉鸡自身不能合成、必须通过饲粮供给的必需氨基酸,但肉鸡常规饲料原料缺乏异亮氨酸,需要额外补充异亮氨酸以达到实际生产需要。基于以上,研究中速型黄羽肉鸡异亮氨酸需要量显得尤为重要,但目前异亮氨酸在鸡上的报道主要集中在快速型肉鸡、种鸡和蛋鸡方面,在中速型黄羽肉鸡上的报道除本课题组前期在1~30日龄和31~60日龄上的研究[15-16]外,尚无其他报道。因此,本试验以61~80日龄中速型黄羽肉鸡为研究对象,研究饲粮不同异亮氨酸水平对其生长性能、血浆生化指标、胴体性状、肉品质、空肠氨基酸转运和肝脏脂质代谢相关基因表达量的影响,为中速型黄羽肉鸡精准营养提供参考。

1 材料与方法

1.1 试验材料

1日龄麻黄肉母雏采购于佛山新广农牧有限公司;异亮氨酸由希杰(上海)商贸有限公司提供,纯度≥90%。本动物试验由广东省农业科学院动物科学研究所实验动物伦理委员会审批,伦理批准编号为2024018。

1.2 试验分组设计

选取1 080只初始体重为1 581.67 g的61日龄黄羽肉母鸡,根据体重一致原则随机分为6组,各组分别额外添加0、0.08%、0.16%、0.24%、0.32%和0.40%晶体异亮氨酸,使试验饲粮异亮氨酸水平依次达到0.46%、0.54%、0.62%、0.70%、0.78%和0.86%,每组6个重复,每个重复30只。试验期20 d。所有试验鸡1~60日龄饲喂相同组成及营养水平的饲粮。

1.3 试验饲粮

试验饲粮所用原料主要为玉米、豆粕、花生粕、玉米蛋白粉和小麦,饲粮营养水平参考《黄羽肉鸡营养需要量》(NY/T 3645—2020)中中速型商品鸡饲粮营养需要配制。采用盐酸水解法利用氨基酸分析仪(LA8080,HITACHI,日本)测得各组饲粮实际异亮氨酸水平分别为0.44%、0.56%、0.61%、0.68%、0.79%和0.87%。饲粮组成及营养水平见表1。
表1 饲粮组成及营养水平(风干基础)

Table 1 Composition and nutrient levels of diets (air-dry basis)%

项目
Items		 饲粮异亮氨酸水平Dietary isoleucine level/%
0.46 0.54 0.62 0.70 0.78 0.86
原料Ingredients
玉米Corn 68.54 68.46 68.38 68.30 68.22 68.14
豆粕Soybean meal 12.00 12.00 12.00 12.00 12.00 12.00
花生粕Peanut meal 6.00 6.00 6.00 6.00 6.00 6.00
玉米蛋白粉Corn gluten meal 5.00 5.00 5.00 5.00 5.00 5.00
小麦Wheat 4.50 4.50 4.50 4.50 4.50 4.50
石粉Limestone 1.25 1.25 1.25 1.25 1.25 1.25
磷酸氢钙CaHPO4 0.72 0.72 0.72 0.72 0.72 0.72
食盐NaCl 0.30 0.30 0.30 0.30 0.30 0.30
L-赖氨酸盐酸盐L-lysine·HCl (98.5%) 0.42 0.42 0.42 0.42 0.42 0.42
DL-蛋氨酸DL-methionine (99%) 0.10 0.10 0.10 0.10 0.10 0.10
L-苏氨酸L-threonine (98.5%) 0.11 0.11 0.11 0.11 0.11 0.11
L-缬氨酸L-valine (98%) 0.06 0.06 0.06 0.06 0.06 0.06
L-异亮氨酸L-isoleucine 0.08 0.16 0.24 0.32 0.40
预混料Premix1) 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
营养水平Nutrient levels2)
氮校正表观代谢能AMEn/(MJ/kg) 12.60 12.60 12.60 12.60 12.60 12.60
粗蛋白质CP 15.99 16.01 15.91 15.91 15.64 16.04
钙Ca 0.70 0.70 0.69 0.69 0.69 0.70
可利用磷AP 0.25 0.25 0.25 0.25 0.25 0.25
赖氨酸Lys 0.81 0.84 0.82 0.80 0.83 0.82
蛋氨酸Met 0.36 0.36 0.36 0.36 0.36 0.36
蛋氨酸+胱氨酸Met+Cys 0.65 0.65 0.65 0.65 0.65 0.65
苏氨酸Thr 0.65 0.66 0.62 0.62 0.64 0.63
色氨酸Trp 0.16 0.16 0.16 0.16 0.16 0.16
异亮氨酸Ile 0.44 0.56 0.61 0.68 0.79 0.87

1)预混料为每千克饲粮提供 The premix provided the following per kg of diets:VA 6 000 IU,VB1 2.1 mg,VB2 3 mg,烟酸 nicotinic acid 15 mg,VB6 3.5 mg,VB12 0.01 mg,VD3 500 IU,VE 20 IU,VK3 0.5 mg,生物素 biotin 0.15 mg,泛酸 pantothenic acid 10 mg,叶酸 folic acid 0.55 mg,氯化胆碱 choline chloride 1 000 mg,Fe 80 mg,Cu 7 mg,Mn 60 mg,Zn 65 mg,I 0.6 mg,Se 0.15 mg。

2)营养水平中粗蛋白质(GB/T 6432—2018)、钙(GB/T 6436—2018)以及赖氨酸、苏氨酸和异亮氨酸(GB/T 18246—2019)为实测值,其余均根据《中国饲料成分及营养价值表(2022年第33版)》中各原料营养成分进行计算。In the nutrient levels, CP (GB/T 6432—2018), Ca (GB/T 6436—2018) as well as Lys, Thr and Ile (GB/T 18246—2019) were measured values, while the rest were calculated according to the values of each raw material in the Tables of Feed Composition and Nutritive Values in China (thirty-third edition, 2022).

1.4 饲养管理

试验在广东省农业科学院动物科学研究所试验场开展,试验鸡平养于封闭式鸡舍,地面铺放木屑,每天自由采食颗粒料和饮水,自然通风,24 h持续光照(白天自然光,夜间补光),按照常规饲养程序和免疫流程进行饲养和免疫。试验期间保持鸡舍清洁卫生,每天08:00、12:00和17:30记录鸡舍温湿度情况,记录死亡鸡只数量和重量。

1.5 样品采集

试验结束时,肉鸡空腹12 h后,每重复随机选取接近平均体重的2只鸡,翅静脉采集5 mL全血于含有肝素钠的抗凝管中,2 500×g离心10 min,取上层血浆分装于离心管中做好标记,-80 ℃保存备测相关指标。上述采血后的鸡颈部放血致死,按照《家禽生产性能名词术语和度量计算方法》(NY/T 823—2020)分割屠体、半净膛、全净膛、胸肌、腿肌和腹脂;剖取整个空肠肠段,在中间部位剪取2 cm左右,用生理盐水冲洗干净内容物,滤纸吸干表面水分后装于1.5 mL无酶离心管中做好标记,-80 ℃保存备测氨基酸转运相关基因表达量;在肝脏相同部位剪取2 g左右,分装于1.5 mL无酶离心管中做好标记,-80 ℃保存备测脂质代谢相关基因表达量。

1.6 指标测定及方法

1.6.1 生长性能

试验结束当晚20:00断料供水,统计每重复采食量,次日08:00以重复为单位称重,计算终末体重、平均日增重、平均日采食量、料重比和死亡率。

1.6.2 血浆生化指标

血浆尿酸(UA)、葡萄糖(GLU)、甘油三酯(TG)、总胆固醇(TC)、谷丙转氨酶(ALT)、谷草转氨酶(AST)、肌酸激酶(CK)和乳酸脱氢酶(LDH)含量或活性测定试剂盒均采购于南京建成生物工程研究所,试剂盒型号依次为C012-1-1、A154-1-1、A111-2-1、A110-2-1、C009-1-1、C010-1-1、A32-1-1和A020-1-2,各指标测定严格按照说明书执行。

1.6.3 胴体性状

根据《家禽生产性能名词术语和度量计算方法》(NY/T 823—2020)中屠宰率、半净膛率、全净膛率、胸肌率、腿肌率和腹脂率的计算方法进行计算。

1.6.4 肉品质

称重后的左侧胸肌用于测定屠宰后45 min和24 h肉色L*值、a*值、黄度(b*)值和pH,右侧胸肌用于测定滴水损失和剪切力。以上肉品质指标具体测定方法参考Ma等[17]的方法。

1.6.5 空肠氨基酸转运和肝脏脂质代谢相关基因表达量

采用Trizol法提取空肠/肝脏组织总RNA,使用核酸浓度测定仪(NanoPhotometer,N60 Touch,德国)测定RNA浓度,依据Color Reverse Transcription Kit试剂盒(EZB,Bioscience,美国)进行RNA反转录。反转录后的cDNA样品稀释10倍进行实时荧光定量PCR(RT-PCR),RT-PCR体系为20 μL(cDNA模板4 μL,上游引物1 μL,下游引物1 μL,ChamQ SYBR- qPCR Master Mix 10 μL,ddH2O 4 μL)。RT-PCR具体程序为:95 ℃预变性3 min,95 ℃变性30 s,退火温度下退火30 s,72 ℃延伸30 s,变性至延伸总共40个循环。内参基因β-肌动蛋白(β-actin)和小肽转运载体1(PepT1)、B0系统中性氨基酸转运载体(B0AT)、兴奋性氨基酸转运载体3(EAAT3)、碱性氨基酸转运载体1(CAT1)、过氧化物酶体增殖物激活受体γ(PPARγ)、3-羟基-3-甲基戊二酰辅酶A还原酶(HMGCR)、沉默信息调节因子1(SIRT1)、肉碱棕榈酰转移酶1(CPT1)等引物信息同范秋丽等[16],采用2-ΔΔCt法计算目的基因相对表达量。

1.7 数据统计与分析

试验数据采用SPSS 23.0软件中的one-way ANOVA程序进行单因素方差分析,在差异显著的基础上进行Duncan氏多重比较以及线性(linear)和二次(quadratic)趋势分析,P<0.05为差异显著,结果数据均以平均值和均值标准误(SEM)表示。
同时,对差异显著的生长性能、胴体性状和肉品质指标应用回归分析进行二次曲线拟合,回归方程为:
y=Ax2+Bx+C。
式中:y代表对应指标测定值;x代表饲粮异亮氨酸水平;A、B和C分别为方程的二次项系数、一次项系数和常数项;应用一次项和二次项系数计算饲粮异亮氨酸适宜水平x0=-B/2A。

2 结果

2.1 饲粮异亮氨酸水平对61~80日龄黄羽肉鸡生长性能的影响

由表2可知,肉鸡料重比随饲粮异亮氨酸水平的升高呈线性和二次变化(P<0.05)。饲粮异亮氨酸水平对肉鸡终末体重、平均日增重、平均日采食量和死亡率均无显著影响(P>0.05)。
表2 饲粮异亮氨酸水平对61~80日龄黄羽肉鸡生长性能的影响

Table 2 Effects of dietary isoleucine level on growth performance of yellow-feathered chickens aged 61 to 80 days

项目
Items		 饲粮异亮氨酸水平Dietary isoleucine level/% SEM P值P-value
0.46 0.54 0.62 0.70 0.78 0.86 方差分析
ANOVA		 线性
Linear		 二次
Quadratic
初始体重
Initial BW/g		 1 581.67 1 581.67 1 581.67 1 581.67 1 581.67 1 581.67 <0.001
终末体重
Final BW/g		 2 138.76 2 148.67 2 152.67 2 158.33 2 147.43 2 131.75 15.483 0.877
平均日增重
ADG/g		 27.86 28.35 28.55 28.83 28.29 27.71 0.709 0.879
平均日采食量
ADFI/g		 110.35 109.95 109.00 106.06 106.05 106.20 1.794 0.083
料重比F/G 3.97a 3.90ab 3.83ab 3.70b 3.78ab 3.80ab 0.102 0.041 0.037 0.013
死亡率Mortality/% 0.56 0.00 1.11 0.00 1.11 1.67 0.300 0.090

同行数据肩标相同字母或无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)。表3至表7同。

In the same row, data shoulder with the same letter or no letter mean no significant difference (P>0.05), while with no identical letter mean significant difference (P<0.05). The same as Table 3 to Table 7.

2.2 饲粮异亮氨酸水平对80日龄黄羽肉鸡血浆生化指标的影响

由表3可知,饲粮异亮氨酸水平显著影响肉鸡血浆GLU和TG含量以及ALT和CK活性(P<0.05),且随异亮氨酸水平的升高,血浆ALT和CK活性均呈线性和二次变化(P<0.05)。与0.46%异亮氨酸水平组相比,0.54%和0.70%异亮氨酸水平组血浆GLU含量显著升高(P<0.05);与0.78%异亮氨酸水平组相比,0.62%、0.70%和0.86%异亮氨酸水平组血浆TG含量显著降低(P<0.05)。饲粮异亮氨酸水平对血浆UA和TC含量以及AST和LDH活性无显著影响(P>0.05)。
表3 饲粮异亮氨酸水平对80日龄黄羽肉鸡血浆生化指标的影响

Table 3 Effects of dietary isoleucine level on plasma biochemical indices of 80-day-old yellow-feathered chickens

项目
Items		 饲粮异亮氨酸水平Dietary isoleucine level/% SEM P值P-value
0.46 0.54 0.62 0.70 0.78 0.86 方差分析
ANOVA		 线性
Linear		 二次
Quadratic
尿酸UA/(mg/L) 90.79 80.17 85.78 91.56 81.70 77.63 2.993 0.718
葡萄糖
GLU/(mmol/L)		 12.47c 13.80a 12.59c 13.57ab 13.23abc 12.78bc 0.135 0.008 0.696 0.367
甘油三酯
TG/(mmol/L)		 0.78ab 0.79ab 0.68b 0.65b 1.05a 0.53b 0.048 0.026 0.735 0.055
总胆固醇
TC/(mmol/L)		 3.75 4.24 3.92 3.54 3.73 3.04 0.132 0.148
谷丙转氨酶
ALT/(U/L)		 2.57b 4.33ab 5.89ab 7.33a 8.72a 7.43a 0.646 0.047 0.002 0.008
谷草转氨酶
AST/(U/L)		 32.13 34.74 37.19 36.14 37.26 38.03 0.892 0.442
肌酸激酶
CK/(U/mL)		 1.56a 1.44ab 1.56a 1.11b 1.20b 1.20b 0.051 0.012 0.003 0.012
乳酸脱氢酶
LDH/(U/L)		 4 227.87 4 067.80 4 082.86 3 818.27 3 940.68 3 920.90 66.552 0.630

2.3 饲粮异亮氨酸水平对80日龄黄羽肉鸡胴体性状的影响

由表4可知,肉鸡腹脂率随饲粮异亮氨酸水平的升高呈线性降低(P<0.05)。饲粮异亮氨酸水平对肉鸡屠宰率、半净膛率、全净膛率、胸肌率和腿肌率均无显著影响(P>0.05)。
表4 饲粮异亮氨酸水平对80日龄黄羽肉鸡胴体性状的影响

Table 4 Effects of dietary isoleucine level on carcass traits of 80-day-old yellow-feathered chickens%

项目
Items		 饲粮异亮氨酸水平Dietary isoleucine level/% SEM P值P-value
0.46 0.54 0.62 0.70 0.78 0.86 方差分析
ANOVA		 线性
Linear		 二次
Quadratic
屠宰率Dressing rate 89.85 89.69 90.14 89.67 89.98 89.48 0.120 0.665
半净膛率
Half-eviscerated rate		 83.43 82.99 83.12 82.99 83.04 83.17 0.153 0.972
全净膛率
Eviscerated rate		 70.27 70.18 69.05 69.38 69.37 69.29 0.212 0.425
胸肌率
Breast muscle rate		 18.36 17.93 18.64 18.22 18.08 18.25 0.423 0.668
腿肌率
Leg muscle rate		 20.93 20.53 20.51 20.04 20.19 20.63 0.187 0.825
腹脂率
Abdominal fat rate		 5.60a 5.51ab 5.54ab 4.82ab 4.55b 4.32b 0.182 0.030 0.029 0.067

2.4 饲粮异亮氨酸水平对80日龄黄羽肉鸡肉品质的影响

由表5可知,饲粮异亮氨酸水平显著影响肉鸡胸肌24 h L*值和a*值以及剪切力(P<0.05),且随异亮氨酸水平的升高,24 h a*值呈线性和二次变化(P<0.05),剪切力呈线性降低(P<0.05)。与0.46%异亮氨酸水平组相比,0.70%和0.78%异亮氨酸水平组胸肌24 h L*值显著降低(P<0.05)。饲粮异亮氨酸水平对肉鸡胸肌45 min肉色(L*值、a*值和b*值)和pH、24 h b*值和pH以及滴水损失均无显著影响(P>0.05)。
表5 饲粮异亮氨酸水平对80日龄黄羽肉鸡肉品质的影响

Table 5 Effects of dietary isoleucine level on meat quality of 80-day-old yellow-feathered chickens

项目
Items		 饲粮异亮氨酸水平Dietary isoleucine level/% SEM P值P-value
0.46 0.54 0.62 0.70 0.78 0.86 方差分析
ANOVA		 线性
Linear		 二次
Quadratic









肉色
Meat
color




45 min		 亮度
L*		 51.93 51.82 52.41 52.71 51.96 51.97 0.418 0.992
红度
a*		 10.86 10.49 9.51 9.83 10.43 9.67 0.157 0.087
黄度
b*		 10.87 11.26 12.20 11.99 12.29 12.48 0.286 0.539





24 h		 亮度
L*		 64.55a 63.17ab 62.64ab 60.87b 60.73b 63.07ab 0.395 0.021 0.828 0.357
红度
a*		 7.93b 8.56b 8.77ab 9.81a 8.90ab 8.29b 0.166 0.028 0.015 0.031
黄度
b*		 19.80 21.08 21.76 20.76 20.69 21.85 0.252 0.168


pH		 45 min 5.65 5.67 5.74 5.71 5.67 5.69 0.017 0.882
24 h 5.58 5.51 5.28 5.53 5.50 5.50 0.027 0.061
滴水损失
Drip loss/%		 2.94 2.56 2.79 2.49 2.54 2.90 0.074 0.263
剪切力
Shear force/N		 35.47a 33.80ab 29.69abc 27.30c 28.96c 26.19c 1.128 0.034 0.001 0.369

2.5 饲粮异亮氨酸水平对80日龄黄羽肉鸡空肠氨基酸转运相关基因表达量的影响

由表6可知,肉鸡空肠EAAT3基因表达量随饲粮异亮氨酸水平的升高呈线性和二次变化(P<0.05)。饲粮异亮氨酸水平对空肠PepT1、B0AT和CAT1基因表达量均无显著影响(P>0.05)。
表6 饲粮异亮氨酸水平对80日龄黄羽肉鸡空肠氨基酸转运相关基因表达量的影响

Table 6 Effects of dietary isoleucine level on jejunal amino acid transport related gene expression of 80-day-old yellow-feathered chickens

项目
Items		 饲粮异亮氨酸水平Dietary isoleucine level/% SEM P值P-value
0.46 0.54 0.62 0.70 0.78 0.86 方差分析
ANOVA		 线性
Linear		 二次
Quadratic
小肽转运载体1
PepT1		 1.02 1.14 1.17 0.98 0.86 0.79 0.075 0.924
B0系统中性氨基
酸转运载体
B0AT		 1.11 1.12 1.15 0.93 1.03 1.07 0.086 0.751
兴奋性氨基酸
转运载体3
EAAT3		 1.06a 0.86ab 0.70abc 0.57bc 0.71abc 0.50c 0.035 0.027 0.001 0.004
碱性氨基酸
转运载体1
CAT1		 1.02 1.01 1.26 1.06 1.18 0.96 0.064 0.921

2.6 饲粮异亮氨酸水平对80日龄黄羽肉鸡肝脏脂质代谢相关基因表达量的影响

由表7可知,随饲粮异亮氨酸水平的升高,肉鸡肝脏HMGCR基因表达量显著降低(P<0.05),同时SIRT1基因表达量显著升高(P<0.05)。饲粮异亮氨酸水平对肝脏PPARγ和CPT1基因表达量无显著影响(P>0.05)。
表7 饲粮异亮氨酸水平对80日龄黄羽肉鸡肝脏脂质代谢相关基因表达量的影响

Table 7 Effects of dietary isoleucine level on liver fat metabolism related gene expression of 80-day-old yellow-feathered chickens

项目
Items		 饲粮异亮氨酸水平Dietary isoleucine level/% SEM P值P-value
0.46 0.54 0.62 0.70 0.78 0.86 方差分析
ANOVA		 线性
Linear		 二次
Quadratic
过氧化物酶体增殖
物激活受体γ
PPARγ		 1.22 1.16 1.14 1.10 1.06 1.09 0.066 0.806
3-羟基-3-甲基
戊二酰辅酶A
还原酶
HMGCR		 1.26a 1.22a 1.13ab 0.74b 0.83b 0.80b 0.058 0.034 0.081 0.060
沉默信息调节因子1
SIRT1		 1.04b 0.96b 1.18ab 1.62a 1.41a 1.28ab 0.087 0.023 0.098 0.146
肉碱棕榈酰转移酶1
CPT1		 1.12 1.17 1.26 1.21 1.10 1.16 0.053 0.520

2.7 回归模型估测饲粮异亮氨酸适宜水平

由表8可知,以料重比、腹脂率、胸肌24 h L*值和a*值以及剪切力为评价指标,通过拟合二次回归方程得出,61~80日龄中速型黄羽肉母鸡饲粮异亮氨酸适宜水平分别为0.74%、0.86%、0.72%、0.68%和0.90%。
表8 回归模型估测饲粮异亮氨酸适宜水平

Table 8 Estimation of optimal level of dietary isoleucine based on regression model

项目
Items		 回归方程
Regression equations		 P值
P-value		 决定系数
R2		 饲粮异亮氨酸适宜水平
Optimal dietary isoleucine level/%
料重比F/G y=2.930x2-4.346x+5.367 0.013 0.886 0.74
腹脂率
Abdominal fat rate/%		 y=-2.297x2+3.931x+6.108 0.067 0.724 0.86
24 h亮度L* at 24 h y=53.902x2-77.324x+88.009 0.357 0.321 0.72
24 h红度a* at 24 h y=-30.554x2+41.833x-5.030 0.031 0.864 0.68
剪切力Shear force/N y=45.553x2-82.245x+26.893 0.369 0.296 0.90

3 讨论

3.1 饲粮异亮氨酸水平对61~80日龄黄羽肉鸡生长性能的影响

研究表明,相同性别肉鸡对支链氨基酸的需要受饲粮组成、肉鸡品种和日龄等因素影响[18]。蒋守群等[9]应用玉米、豆粕和血球蛋白粉组成的饲粮发现,0.55%~0.90%、0.50%~0.85%和0.45%~0.80%异亮氨酸分别显著影响快大型黄羽肉母鸡1~21日龄平均日增重、平均日采食量和料重比,22~42日龄平均日增重和料重比,以及43~63日龄平均日增重、平均日采食量和料重比,且随饲粮异亮氨酸水平的升高,1~21日龄和43~63日龄料重比均呈线性和二次变化,22~42日龄料重比呈二次变化。Hale等[19]使用玉米和豆粕等组成的饲粮,分别在30~42日龄爱拔益加肉母鸡、罗斯508和罗斯708肉母鸡上研究发现,0.42%~0.82%异亮氨酸显著影响体重、采食量和料重比,且异亮氨酸对体重和采食量的影响存在品种效应。本课题组前期使用玉米、小麦、花生粕和豆粕组成的饲粮在中速型黄羽肉母鸡上的研究表明,随着1~30日龄肉鸡饲粮异亮氨酸从0.68%升高到1.08%,平均日增重和饲料转化率均显著升高,且呈线性和二次变化[15];同时,31~60日龄肉鸡料重比随饲粮异亮氨酸水平从0.58%升高到0.98%呈线性和二次曲线降低[16]。本研究中,饲粮0.46%~0.86%水平异亮氨酸虽然对61~80日龄肉鸡体重、平均日增重和平均日采食量无显著影响,但随异亮氨酸水平的升高,料重比呈线性和二次变化,此结果与前人研究结果一致,说明适宜水平的异亮氨酸对肉鸡生长有促进作用;此外,以料重比为评价指标,二次回归模型估测饲粮异亮氨酸适宜水平为0.74%,此水平高于《黄羽肉鸡营养需要量》(NY/T 3645—2020)中相同日龄阶段中速型黄羽肉母鸡异亮氨酸水平0.57%,也高于《鸡饲养标准》(NY/T 33—2004)中相似日龄阶段肉仔鸡异亮氨酸水平0.63%,其原因与肉鸡遗传性状的改善提高了氨基酸需要量有关。

3.2 饲粮异亮氨酸水平对80日龄黄羽肉鸡血浆生化指标的影响

血液生化指标可在一定程度反映机体生理机能和营养物质代谢情况。GLU可直接为机体提供能量,其含量升高,表明机体对饲粮中碳水化合物的利用提高[20]。本研究中,肉鸡血浆GLU含量随饲粮异亮氨酸水平的升高而有所升高,此结果虽与郭建来等[21]研究异亮氨酸水平对产蛋后期蛋鸡血清GLU含量无影响的结果不同,但与Lin等[22]在黄羽肉种鸡上的研究相似,43和45周龄血浆GLU含量随异亮氨酸水平的升高而升高。TG含量升高,表明机体脂肪合成增加[23]。尚晓迪等[24]研究表明,草鱼幼鱼血清TG含量随饲粮异亮氨酸水平的升高呈先降低后升高的趋势,本研究结果与其相似,肉鸡血浆TG含量随饲粮异亮氨酸水平的升高同样呈先降低后升高的趋势。血液ALT活性是检测肝脏功能受损程度的常用指标[25],CK主要存在于骨骼肌和心肌中,在骨骼肌收缩和胞内能量转换中起到重要作用[26]。Lin等[22]研究表明,43和45周龄黄羽肉种鸡血浆CK活性随饲粮异亮氨酸水平的升高显著降低,这与本研究中肉鸡血浆CK活性随饲粮异亮氨酸水平的升高呈线性降低的结果一致;此外,本研究还发现,肉鸡血浆ALT活性随饲粮异亮氨酸水平的升高呈线性和二次升高,说明异亮氨酸的添加虽然降低了肉鸡心肌和骨骼肌损伤的风险,但同时也可能对肝脏造成损伤。

3.3 饲粮异亮氨酸水平对80日龄黄羽肉鸡胴体性状的影响

与其他肉类产品相比,禽肉以其高蛋白质、低脂肪含量更受消费者青睐。Maynard等[27]研究发现,缬氨酸、亮氨酸和异亮氨酸互作虽然对44日龄科宝肉鸡屠宰率、胸肌率和腿肌率无影响,但显著影响腹脂率,且随异亮氨酸水平从0.62%升高到0.70%,腹脂率显著降低。李守学等[28]研究表明,2%和5%异亮氨酸和同等水平的精氨酸混合添加可降低了21日龄罗斯308肉鸡腹脂率,但对屠宰率、半净膛率、全净膛率、胸肌率和腿肌率无影响。本研究中,随饲粮异亮氨酸水平从0.46%升高到0.86%,80日龄黄羽肉母鸡腹脂率呈线性降低,屠宰率、半净膛率、全净膛率、胸肌率和腿肌率均不受影响,此结果与丁元翠等[29]研究相似,不同水平异亮氨酸对90日龄皖西白鹅屠宰率、半净膛率、全净膛率、胸肌率和腿肌率无影响;但与本课题组前期报道[16]不尽一致,0.58%~0.98%异亮氨酸降低60日龄黄羽肉鸡腹脂率的同时也提高了屠宰率、半净膛率和全净膛率,其原因可能与随着鸡日龄的增加,异亮氨酸对胴体蛋白质沉积的影响不如赖氨酸和蛋氨酸等其他主要氨基酸显著有关[30-31]。

3.4 饲粮异亮氨酸水平对80日龄黄羽肉鸡肉品质的影响

肉色和剪切力可分别作为评价肉品新鲜度和嫩度的重要表观指标。刘恒晨等[11]研究表明,适宜水平的异亮氨酸可提高42日龄肉鸡屠宰后45 min胸肌和腿肌a*值。本研究中,随着饲粮异亮氨酸水平的升高,80日龄肉鸡屠宰后24 h胸肌L*值降低,a*值升高,且后者呈线性和二次变化,剪切力呈线性降低,这与罗燕红等[32]研究肥育猪背最长肌剪切力随异亮氨酸水平的升高呈线性降低的结果相似,但缺乏或者过量的异亮氨酸对L*值和a*值无影响。李洪涛等[13]研究也发现,适宜水平的异亮氨酸和精氨酸混合添加可提高42日龄肉鸡屠宰后45 min胸肌a*值,降低腿肌L*值。上述结果表明,适宜水平的异亮氨酸可通过改善肉色、降低剪切力提高黄羽肉鸡肉品质。

3.5 饲粮异亮氨酸水平对80日龄黄羽肉鸡空肠氨基酸转运和肝脏脂质代谢相关基因表达的影响

EAAT3是小肠内酸性氨基酸的主要转运载体,其表达量升高,说明小肠对酸性氨基酸的转运和吸收增加[33]。本课题组前期研究发现,饲粮异亮氨酸水平对60日龄黄羽肉鸡空肠和回肠EAAT3基因表达量均无显著影响[16]。本研究中,80日龄黄羽肉鸡空肠EAAT3基因表达量随饲粮异亮氨酸水平的升高呈线性和二次降低,这可能与支链氨基酸之间相互竞争从而降低了EAAT3的表达有关[34]。SIRT1是一种主要存在于肝脏、心脏、肌肉和脂肪组织中的脱乙酰酶,其通过阻碍脂肪酸氧化和促进白色脂肪组织棕色化过程降低脂肪沉积,具体表现为如下调脂肪合成相关基因PPARγ和HMGCR表达量以及上调脂肪分解相关基因CPT1表达量[35-36]。本研究中,随着饲粮异亮氨酸水平的升高,肉鸡肝脏HMGCR基因表达量降低、SIRT1基因表达量升高,此结果与本课题组前期在60日龄黄羽肉鸡肝脏上的研究结果[16]一致,同时也与本文中异亮氨酸对腹脂率和血浆TG含量影响的结果相呼应,说明适宜水平的异亮氨酸可通过调节脂质代谢相关基因表达和降低血脂含量起到降低体脂沉积的作用。

4 结论

饲粮适宜水平的异亮氨酸可改善61~80日龄中速型黄羽肉母鸡生长性能、胴体性状和肉品质,同时调节血糖和血脂含量以及空肠兴奋性氨基酸转运载体基因和肝脏脂质代谢相关基因表达。依据二次回归模型估测,61~80日龄中速型黄羽肉母鸡获得最佳生长性能、胴体性状和肉品质的饲粮异亮氨酸水平为0.68%~0.90%,其中以料重比为主要评价指标,饲粮异亮氨酸适宜水平为0.74%,每日需要量为742 mg。

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