研究论文

N-酰基高丝氨酸内酯酶对肉仔鸡血清生化、免疫指标以及肠道形态的影响

  • 郑爱娟 ,
  • 陈星 ,
  • 张广民 ,
  • 王泽栋 ,
  • 陈志敏 ,
  • 常文环 ,
  • 蔡辉益 ,
  • 刘国华 , *
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  • 中国农业科学院饲料研究所,农业农村部动物产品质量安全饲料源性因子风险评估实验室,农业农村部饲料生物技术重点实验室,北京 100081
* 刘国华,研究员,博士生导师,E-mail:

郑爱娟(1977—),女,河北唐山人,副研究员,博士,从事家禽分子营养和家禽生态养殖营养调控技术研究。E-mail:

Office editor: 陈 鑫

收稿日期: 2022-12-21

  网络出版日期: 2023-07-11

基金资助

国家肉鸡产业技术体系(CARS-41)

Effects of N-Acylhomoserine Lactone Enzyme on Serum Biochemical, Immune Indexes and Intestinal Morphology of Broiler Chickens

  • ZHENG Aijuan ,
  • CHEN Xing ,
  • ZHANG Guangmin ,
  • WANG Zedong ,
  • CHEN Zhimin ,
  • CHANG Wenhuan ,
  • CAI Huiyi ,
  • LIU Guohua , *
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  • Key Laboratory of Feed Biotechnology of the Ministry of Agriculture and Rural Affairs, Risk Assessment Laboratory of Animal Product Quality Safety Feed Source Factors of the Ministry of Agriculture and Rural Affairs, Feed Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081, China
* professor, E-mail:

Received date: 2022-12-21

  Online published: 2023-07-11

摘要

本试验旨在研究N-酰基高丝氨酸内酯酶对肉仔鸡血清生化、免疫指标和肠道形态的影响。选取1日龄健康、体重相近的科宝肉仔鸡600只,随机分为5组,每组6个重复,每个重复20只鸡。对照组饲喂基础饲粮,试验组在基础饲粮中分别添加250、500、750和1 000 mg/kg的N-酰基高丝氨酸内酯酶。试验期42 d。结果表明:1)与对照组相比,饲粮中添加N-酰基高丝氨酸内酯酶显著提高21和42日龄肉鸡血清中总蛋白、白蛋白、球蛋白和尿酸含量,以及21日龄血清尿素氮含量(P<0.05)。2)与对照组相比,饲粮中添加N-酰基高丝氨酸内酯酶显著提高21日龄肉鸡血清中碱性磷酸酶活性(P<0.05)。3)与对照组相比,饲粮中添加N-酰基高丝氨酸内酯酶显著提高21日龄肉鸡血清中脂蛋白含量,以及42日龄血清脂蛋白和胆固醇含量(P<0.05)。4)与对照组相比,饲粮中添加N-酰基高丝氨酸内酯酶显著提高21和42日龄肉鸡血清中免疫球蛋白A和免疫球蛋白M含量(P<0.05)。5)与对照组相比,饲粮中添加N-酰基高丝氨酸内酯酶显著提高肉鸡的胰腺指数(P<0.05)。6)与对照组相比,饲粮中添加N-酰基高丝氨酸内酯酶显著提高肉鸡十二指肠、空肠和回肠的绒毛高度和绒隐比(P<0.05),显著降低回肠隐窝深度(P<0.05)。综上所述,饲粮中添加N-酰基高丝氨酸内酯酶促进蛋白质和脂质在肉鸡体内的代谢和周转,并改善了肠道形态,其中添加500 mg/kg的N-酰基高丝氨酸内酯酶对肉鸡健康和生长的效果最佳。

本文引用格式

郑爱娟 , 陈星 , 张广民 , 王泽栋 , 陈志敏 , 常文环 , 蔡辉益 , 刘国华 . N-酰基高丝氨酸内酯酶对肉仔鸡血清生化、免疫指标以及肠道形态的影响[J]. 动物营养学报, 2023 , 35(7) : 4301 -4311 . DOI: 10.12418/CJAN2023.400

Abstract

The purpose of this experiment was to study the effects of N-acylhomoserine lactone enzyme on serum biochemical, immune indexes and intestinal morphology of broiler chickens. A total of 600 healthy 1-day-old Cobb broiler chickens were randomly divided into 5 groups, with 6 replicates in each group and 20 chickens in each replicate. The broilers in control group were fed a basic diet, and the those in test groups were fed the basic diet supplemented with 250, 500, 750, and 1 000 mg/kg of N-acylhomoserine lactone enzyme, respectively. The test period was 42 days. The results showed as follows: 1) the contents of total protein, albumin, globulin and uric acid in serum of broilers at 21 and 42 days of age, and the content of urea nitrogen in serum at 21 days of age were significantly increased by adding N-acylhomoserine lactone enzyme to the diet (P<0.05). 2) Addition of N-acylhomoserine lactone enzyme to the diet significantly increased the serum alkaline phosphatase activity of 21-day-old broilers (P<0.05). 3) The addition of N-acylhomoserine lactone enzyme to the diet significantly increased the content of lipoprotein in serum of 21-day-old broilers, and the contents of lipoprotein and cholesterol in serum of 42-day-old broilers (P<0.05). 4) The addition of N-acylhomoserine lactone enzyme to the diet significantly increased the contents of immune globulin (Ig) A and IgM in serum of 21 and 42-day-old broilers (P<0.05). 5) The diet supplemented with N-acylhomoserine lactone enzyme significantly increased the pancreatic index of broilers (P<0.05). 6) Addition of N-acylhomoserine lactone enzyme to the diet significantly increased the villus height to villus crypt ratio of duodenum, jejunum and ileum in broilers (P<0.05), and significantly decreased the depth of ileum crypt (P<0.05). In conclusion, the addition of N-acylhomoserine lactone enzyme in the diet enhances the metabolism and turnover of protein and lipid in broilers, and improves the intestinal morphology. The effect of adding 500 mg/kg N-acylhomoserine lactone enzyme to the diets on the health and growth of broilers is optimal.

群体感应(quorum sensing, QS)是细菌通过释放自诱导信号分子(autoinducers, AIs)来调控相关基因的表达,进而影响毒素合成、黏附和生物被膜的形成等生物学功能的群体行为[1-2]。细菌中存在多种由不同种类AIs介导的QS系统,病原菌的致病作用与QS系统密切相关[3-4]。但是,群体感应淬灭(quorum quenching, QQ)是对QS系统的干预和破坏,通过QQ阻止AIs的积累,干扰其信号传递及其与受体的识别和结合,阻断细菌间的信息交流,抑制毒力基因表达,降低细菌致病性[5]。N-酰基高丝氨酸内酯(AHL)是研究最透彻的介导革兰氏阴性菌QS系统中的信号分子。有超过25种革兰氏阴性菌QS系统都是由AHL进行调节的[6]。随着细菌浓度增加,AHL浓度超过一定阈值水平时,与相应受体蛋白结合来调控细菌的多种生理功能,如溶血素等毒力因子分泌、生物发光和色素生产等[7]。群体感应淬灭酶能够降解AHL,使其无法达到引发致病因子表达的临界浓度,病原菌就失去致病力。N-酰基高丝氨酸内酯酶(AHL酶)主要作用于高丝氨酸内酯环,从五环中酯键碳与氧之间打开内酯环,生物降解信号分子AHL,阻断病原菌的QS机制,从而抑制病原菌的致病性[8]。这种淬灭酶不作用于致病菌本身,而是作用于致病菌产生的信号分子,不会使致病菌产生抗药性,是一种高效、安全的抗菌生物制剂。AHL酶活性高,具有较强的蛋白酶抗性,可作为新型的饲料添加剂应用于畜禽饲料。前期研究发现,饲粮中添加AHL酶显著降低肉鸡的平均日采食量和料重比。饲喂250~500 mg/kg AHL酶显著提高肉鸡对饲粮中干物质、粗蛋白质和能量的表观代谢率[9]。本试验研究AHL酶对肉鸡血清生化指标、免疫指标和肠道形态的影响,探究AHL酶改善肉鸡生长发育的机制,为AHL酶在肉鸡生产上的应用提供理论依据。

1 材料与方法

1.1 试验材料

AHL酶外观性状为乳白色粉末,活性为104 U/g,由北京某生物技术有限公司惠赠。

1.2 试验设计

采用单因素试验设计,选取健康、体重相近的1日龄科宝肉仔鸡600只,随机分为5组,每组6个重复,每个重复20只鸡。肉仔鸡初始平均体重为(46.15±0.45) g,保证各组间体重差异不显著(P>0.05)。对照组饲喂基础饲粮,试验组在基础饲粮中分别添加250、500、750和1 000 mg/kg的AHL酶。基础饲粮参考NY/T 33—2004[10]进行配制,基础饲粮组成及营养水平见文献[9]。肉鸡的光照和黑暗时长分别为16和8 h,使用乳头式饮水器供水,自由采食。使用水循环式暖气进行供热,前3 d鸡舍温度维持在33 ℃,此后每周降低2 ℃,直到维持在24 ℃。在1日龄时接种马立克疫苗,7日龄时滴鼻点眼、接种新城疫、肾传支二联苗。试验鸡饲养管理、免疫规程和鸡舍卫生管理均按照科宝肉鸡饲养规程进行,试验期42 d。

1.3 指标测定

1.3.1 血清生化指标

于42日龄时,每个重复取2只鸡,心脏采血约10 mL,制备血清,采用相应试剂盒(南京建成生物工程研究所)在全自动生化分析仪(日立7160)上测定血清谷丙转氨酶(ALT)、谷草转氨酶(AST)和碱性磷酸酶(ALP)活性,以及血清总蛋白(TP)、白蛋白(ALB)、球蛋白(GLB)、尿素氮(UREA)、肌酐(CREA)、甘油三酯(TG)、脂蛋白(LPA)、总胆固醇(TC)含量。

1.3.2 血清免疫指标

采用免疫比浊试剂盒(南京建成生物工程研究所)测定血清免疫球蛋白A(IgA)、免疫球蛋白M(IgM)和免疫球蛋白G(IgG)含量。采用红血球凝集抑制法测定血清新城疫抗体滴度。

1.3.3 器官指数

于42日龄,每个重复选取2只鸡,采集胰腺、脾脏、胸腺和法氏囊,生理盐水清洗后,使用吸水纸擦拭干称重,计算胰腺、脾脏、胸腺和法氏囊重与活重的比值,即器官指数。

1.3.4 肠道形态

十二指肠、空肠和回肠标本用4%多聚甲醛固定,按照SOP程序进行修剪、脱水、包埋、切片、染色和密封。使用Case Viewer scanning browser software (Case Viewer 2.2,匈牙利)选择组织的目标区域。拍照和成像后,使用Image Pro Plus 6.0分析软件(Image-Pro Plus 6.0,美国)测量切片的绒毛高度(VH)和隐窝深度(CD),并计算绒隐比(V/C)。

1.4 统计分析

所有数据均采用SAS 9.4中的一般线性模型(GLM)进行分析,对主效应显著的指标采用Tukey’s法进行多重比较。结果用平均值和均值标准误(SEM)表示。在所有分析中,P<0.05为差异显著。

2 结果与分析

2.1 AHL酶对肉鸡血清生化指标的影响

2.1.1 AHL酶对肉鸡血清蛋白及其代谢物含量的影响

表2可知,在21日龄,与对照组相比,250 mg/kg组和1 000 mg/kg组肉鸡血清中总蛋白含量显著降低(P<0.05),但组间无显著差异(P>0.05);500 mg/kg组血清中总蛋白含量显著高于对照组(P<0.05),而750 mg/kg组则与对照组无显著差异(P>0.05)。500 mg/kg组肉鸡血清中白蛋白含量显著高于250 mg/kg组(P<0.05),但均与对照组无显著差异(P>0.05)。对照组、250 mg/kg组、750 mg/kg组和1 000 mg/kg组之间血清球蛋白含量无显著差异(P>0.05),但显著低于500 mg/kg组(P<0.05)。与对照组相比,750 mg/kg组和1 000 mg/kg组血清中肌酐含量显著提高(P<0.05),其中750 mg/kg组显著高于1 000 mg/kg组(P<0.05);250 mg/kg组和500 mg/kg组与对照组之间无显著差异(P>0.05)。血清尿素氮含量在1 000 mg/kg组表现出最高水平,而对照组、250 mg/kg组、500 mg/kg组和750 mg/kg组之间无显著差异(P>0.05)。
表2 饲粮中添加AHL酶对肉鸡血清蛋白及其代谢物含量的影响

Table 2 Effects of dietary supplementation of AHL enzyme on serum protein and its metabolite contents of broiler chickens

项目
Items
总蛋白
TP/
(g/L)
白蛋白
ALB/
(g/L)
球蛋白
GLB/
(g/L)
肌酐
CREA/
(μmol/L)
尿素氮
UREA/
(mmol/L)
21日龄21 days of age
对照组Control group 27.13b 11.93ab 15.21b 24.63c 0.61b
250 mg/kg组250 mg/kg group 23.93c 10.17b 13.77b 33.33bc 0.64b
500 mg/kg组500 mg/kg group 31.17a 12.77a 18.41a 31.87bc 0.67b
750 mg/kg组750 mg/kg group 26.83b 11.67ab 15.17b 50.01a 0.68b
1 000 mg/kg组1 000 mg/kg group 21.41c 11.81ab 12.93b 39.17b 0.81a
SEM 0.933 0.328 0.615 2.528 0.021
PP-value <0.001 0.037 0.021 0.002 0.007
42日龄42 days of age
对照组Control group 30.01bc 12.23bc 17.67cd 23.10b 0.74
250 mg/kg组250 mg/kg group 28.63c 11.81c 16.01d 25.21ab 0.74
500 mg/kg组500 mg/kg group 32.07ab 12.97ab 22.07a 33.37a 0.72
750 mg/kg组750 mg/kg group 31.37b 13.51a 18.67bc 33.07a 0.71
1 000 mg/kg组1 000 mg/kg group 33.73a 12.91ab 20.83ab 30.87ab 0.70
SEM 0.536 0.188 0.642 1.458 0.031
PP-value 0.004 0.008 0.001 0.044 0.225

同列数据肩标不同小写字母表示差异显著(P<0.05),相同字母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.

在42日龄,与对照组相比,1 000 mg/kg组血清中总蛋白含量显著提高(P<0.05),750 mg/kg组显著高于250 mg/kg组(P<0.05),但与对照组和500 mg/kg组之间无显著差异(P>0.05);750 mg/kg组血清白蛋白含量显著高于对照组和250 mg/kg组(P<0.05),但与500 mg/kg组和1 000 mg/kg组无显著差异(P>0.05);500 mg/kg组血清球蛋白含量显著高于对照组、250 mg/kg组和750 mg/kg组(P<0.05),但与1 000 mg/kg组无显著差异(P>0.05);对照组血清中肌酐含量显著低于500 mg/kg组和750 mg/kg组(P<0.05),但与250 mg/kg组和1 000 mg/kg组之间无显著差异(P>0.05)。

2.1.2 AHL酶对肉鸡血清酶类指标的影响

表3可知,在21日龄,与对照组相比,250 mg/kg组、500 mg/kg组、750 mg/kg组和1 000 mg/kg组中血清碱性磷酸酶活性显著提高(P<0.05),其中500 mg/kg组显著高于250 mg/kg组、750 mg/kg组和1 000 mg/kg组(P<0.05),但这3组之间无显著差异(P>0.05);饲粮中添加AHL酶对肉鸡血清中谷丙转氨酶和谷草转氨酶活性无显著影响(P>0.05)。
表3 饲粮中添加AHL酶对肉鸡血清酶类指标的影响

Table 3 Effects of dietary supplementation of AHL enzyme on serum enzyme indexes of broiler chickens U/L

项目Items 谷丙转氨酶ALT 谷草转氨酶AST 碱性磷酸酶ALP
21日龄21 days of age
对照组Control group 3.21 126.67 67.61c
250 mg/kg组250 mg/kg group 4.04 127.12 101.64b
500 mg/kg组500 mg/kg group 3.92 128.64 119.63a
750 mg/kg组750 mg/kg group 4.15 129.33 93.12b
1 000 mg/kg组1 000 mg/kg group 4.07 128.97 95.04b
SEM 0.266 3.418 4.745
PP-value 0.384 0.749 <0.001
42日龄42 days of age
对照组Control group 3.59 270.77 37.73
250 mg/kg组250 mg/kg group 3.43 288.60 38.33
500 mg/kg组500 mg/kg group 3.51 272.30 40.83
750 mg/kg组750 mg/kg group 3.62 278.23 37.03
1 000 mg/kg组1 000 mg/kg group 3.60 283.90 44.23
SEM 0.249 8.742 1.859
PP-value 0.226 0.174 0.055
在42日龄,饲粮中添加AHL酶对血清中谷丙转氨酶、谷草转氨酶和碱性磷酸酶活性均无显著影响(P>0.05)

2.1.3 AHL酶对肉鸡血清脂类指标的影响

表4可知,在21日龄,与对照组相比,750 mg/kg组血清中脂蛋白含量显著降低(P<0.05),其他组间无显著差异(P>0.05);饲粮中添加AHL酶对血清总胆固醇和甘油三酯含量无显著影响(P>0.05)。
表4 饲粮中添加AHL酶对肉鸡血清脂类指标的影响

Table 4 Effects of dietary supplementation of AHL enzyme on serum lipid indexes of broiler chickens

项目Items 脂蛋白LPA/(μg/mL) 总胆固醇TC/(mmol/L) 甘油三酯TG/(mmol/L)
21日龄21 days of age
对照组Control group 13.89a 2.77 0.55
250 mg/kg组250 mg/kg group 14.57a 2.62 0.53
500 mg/kg组500 mg/kg group 14.55a 2.72 0.52
750 mg/kg组750 mg/kg group 11.77b 2.91 0.61
1 000 mg/kg组1 000 mg/kg group 13.47a 2.84 0.54
SEM 0.325 0.062 0.016
PP-value 0.008 0.678 0.556
42日龄42 days of age
对照组Control group 23.44a 3.06b 0.56
250 mg/kg组250 mg/kg group 16.99bc 3.02b 0.69
500 mg/kg组500 mg/kg group 22.19a 3.25ab 0.75
750 mg/kg组750 mg/kg group 20.72ab 3.31ab 0.67
1 000 mg/kg组1 000 mg/kg group 15.01c 3.44a 0.67
SEM 1.006 0.053 0.023
PP-value 0.009 0.032 0.097
在42日龄,对照组血清中脂蛋白含量显著高于250 mg/kg组和1 000 mg/kg组(P<0.05),但与500 mg/kg组和750 mg/kg组之间无显著差异(P>0.05);对照组血清总胆固醇含量显著低于1 000 mg/kg组(P<0.05),但与250 mg/kg组、500 mg/kg组和750 mg/kg组之间无显著差异(P>0.05)。

2.2 AHL酶对肉鸡免疫指标的影响

2.2.1 AHL酶对肉鸡血清免疫球蛋白含量和新城疫抗体滴度的影响

表5可知,在21日龄,与对照组相比,750 mg/kg组肉鸡血清中IgA含量显著提高(P<0.05),但与250 mg/kg组和500 mg/kg组之间无显著差异(P>0.05);对照组血清中IgM含量显著低于750 mg/kg组(P<0.05),但与250 mg/kg组、500 mg/kg组和1 000 mg/kg组无显著差异(P>0.05)。
表5 饲粮中添加AHL酶对肉鸡血清免疫球蛋白含量和抗体滴度的影响

Table 5 Effects of dietary supplementation of AHL enzyme on serum immunoglobulin contents and antibody titer of broiler chickens

项目
Items
免疫球蛋白A
IgA/(g/L)
免疫球蛋白M
IgM/(g/L)
免疫球蛋白G
IgG/(g/L)
抗体滴度
Antibody titer
21日龄21 days of age
对照组Control group 0.86bc 0.68b 7.24 8.51
250 mg/kg组250 mg/kg group 0.99ab 0.82b 7.61 8.33
500 mg/kg组500 mg/kg group 0.96abc 0.85b 7.98 8.51
750 mg/kg组750 mg/kg group 1.11a 1.09a 8.07 8.53
1 000 mg/kg组1 000 mg/kg group 0.77c 0.69b 7.51 8.42
SEM 0.039 0.044 0.156 0.257
PP-value 0.023 0.002 0.468 0.694
42日龄42 days of age
对照组Control group 1.08c 0.75b 9.37 8.17
250 mg/kg组250 mg/kg group 1.16bc 0.74b 8.69 7.83
500 mg/kg组500 mg/kg group 1.48a 1.04a 10.12 8.67
750 mg/kg组750 mg/kg group 1.24b 0.95a 9.46 7.41
1 000 mg/kg组1 000 mg/kg group 1.34bc 0.73b 10.11 7.83
SEM 0.039 0.038 0.205 0.551
PP-value 0.002 0.001 0.131 0.314
在42日龄,500 mg/kg组和750 mg/kg组血清中IgA含量显著高于对照组(P<0.05);对照组血清中IgM含量显著低于500 mg/kg组和750 mg/kg组(P<0.05),但与250 mg/kg组和1 000 mg/kg组之间无显著差异(P>0.05)。对照组血清中IgG含量和新城疫抗体滴度与250 mg/kg组、500 mg/kg组、750 mg/kg组和1 000 mg/kg组之间无显著差异(P>0.05)。

2.2.2 AHL酶对肉鸡免疫器官指数的影响

表6可知,与对照组相比,750 mg/kg组肉鸡胰腺指数显著提高(P<0.05)。然而,对照组与250 mg/kg组、500 mg/kg组、750 mg/kg组和1 000 mg/kg组之间脾脏、胸腺和法氏囊指数并无显著差异(P>0.05)。
表6 饲粮中添加AHL酶对肉鸡免疫器官指数的影响

Table 6 Effects of dietary supplementation of AHL enzyme on immune organ indexes of broiler chickens %

项目
Items
胰腺指数
Pancreas index
脾脏指数
Spleen index
胸腺指数
Thymus index
法氏囊指数
Bursa of
Fabricius index
对照组Control group 0.195b 0.099 0.342 0.062
250 mg/kg组250 mg/kg group 0.202ab 0.110 0.290 0.046
500 mg/kg组500 mg/kg group 0.171b 0.114 0.302 0.053
750 mg/kg组750 mg/kg group 0.233a 0.098 0.368 0.055
1 000 mg/kg组1 000 mg/kg group 0.185b 0.099 0.301 0.067
SEM 0.006 0.007 0.019 0.004
PP-value 0.008 0.932 0.789 0.553

2.3 AHL酶对肉鸡肠道形态的影响

表7可见,与对照组相比,500 mg/kg组十二指肠绒毛高度显著提高(P<0.05);对照组绒毛高度显著高于1 000 mg/kg组(P<0.05),但与250 mg/kg组和750 mg/kg组之间无显著差异(P>0.05);500 mg/kg组和750 mg/kg组绒隐比显著高于对照组、250 mg/kg组和750 mg/kg组(P<0.05),对照组、250 mg/kg组和750 mg/kg组间无显著差异(P>0.05)。对照组与250 mg/kg组和750 mg/kg组空肠绒毛高度无显著差异(P>0.05),但显著低于500 mg/kg组并高于1 000 mg/kg组(P<0.05);1 000 mg/kg组绒隐比显著低于对照组、250 mg/kg组和500 mg/kg组(P<0.05),但与750 mg/kg组之间无显著差异(P>0.05)。对照组回肠绒毛高度显著低于250 mg/kg组、500 mg/kg组、750 mg/kg组和1 000 mg/kg组(P<0.05);与对照组相比,250 mg/kg组、500 mg/kg组、750 mg/kg组和1 000 mg/kg组隐窝深度显著降低(P<0.05),但组间无显著差异(P>0.05);与对照组相比,回肠绒隐比显著低于250 mg/kg组、500 mg/kg组、750 mg/kg组和1 000 mg/kg组(P<0.05),其中250 mg/kg组、500 mg/kg组和750 mg/kg组显著高于1 000 mg/kg组(P<0.05),但3组之间无显著差异(P>0.05)。
表7 饲粮中添加AHL酶对肉鸡肠道形态的影响

Table 7 Effects of dietary supplementation of AHL enzyme on intestinal morphology of broiler chickens

项目Items 绒毛高度VH/μm 隐窝深度CD/μm 绒隐比V/C
十二指肠Duodenum
对照组Control group 1 582.33b 394.85 4.01b
250 mg/kg组250 mg/kg group 1 565.89bc 389.30 3.99b
500 mg/kg组500 mg/kg group 1 640.04a 388.20 4.23a
750 mg/kg组750 mg/kg group 1 627.72ab 388.26 4.19a
1 000 mg/kg组1 000 mg/kg group 1 544.09c 386.16 3.99b
SEM 10.725 1.455 0.032
PP-value <0.001 0.442 0.001
空肠Jejunum
对照组Control group 1 252.33b 361.55 3.46a
250 mg/kg组250 mg/kg group 1 254.07b 362.79 3.46a
500 mg/kg组500 mg/kg group 1 571.37a 363.52 3.49a
750 mg/kg组750 mg/kg group 1 244.04b 362.16 3.44ab
1 000 mg/kg组1 000 mg/kg group 1 225.67c 363.30 3.37b
SEM 4.468 0.571 0.014
PP-value 0.002 0.854 0.028
回肠Ileum
对照组Control group 904.92c 290.88a 3.11c
250 mg/kg组250 mg/kg group 1 047.33a 253.29b 4.13a
500 mg/kg组500 mg/kg group 1 048.49a 250.01b 4.20a
750 mg/kg组750 mg/kg group 1 021.72ab 247.18b 4.14a
1 000 mg/kg组1 000 mg/kg group 964.35b 255.25b 3.78b
SEM 14.959 4.345 0.109
PP-value <0.001 <0.001 <0.001

3 讨论

3.1 AHL酶对肉鸡血清生化指标的影响

血清生化指标反映动物机体健康状况。血清总蛋白含量是反映机体肝脏蛋白质合成与代谢以及组织蛋白质吸收利用率的重要指标[11]。总蛋白包含白蛋白和球蛋白,白蛋白比例与肝脏功能损伤程度呈正相关[12],而球蛋白则与机体免疫功能密切相关[13]。本研究中,饲粮中添加AHL酶显著提高了21和42日龄肉鸡血清中总蛋白、白蛋白和球蛋白含量,其中添加500 mg/kg AHL酶组的效果最佳。此外,肌酐作为肉鸡肌肉中肌酸和磷酸肌酸的分解产物,与机体的肌肉总量密切相关[14]。尿素氮是机体蛋白质代谢的主要终产物,经肾小球滤过后随尿液排出,是评价动物肝脏功能的重要指标[15]。饲粮中添加AHL酶显著提高了生长前期(1~21日龄)肉鸡血清中尿素氮含量,表明AHL酶可能提高了肉鸡蛋白质代谢率。饲粮中添加AHL酶显著提高了21以及42日龄时肉鸡血清中肌酐的含量。同时,本试验发现饲粮中添加AHL酶显著提高了21日龄肉鸡血清中脂蛋白含量以及42日龄肉鸡血清脂蛋白和总胆固醇含量。总蛋白、白蛋白、球蛋白和肌酐均是机体蛋白质新陈代谢的关键指标[16]。血清脂蛋白和总胆固醇含量在一定程度上反映肉鸡体内脂类代谢状况[17]。饲粮中添加AHL酶显著改善了21和42日龄肉鸡对蛋白质和脂质的表观代谢率,这可能是通过AHL酶破坏了高丝氨酸内酯环,使信号分子AHL得到降解,阻断病原菌之间的QS机制进而抑制了病原菌的致病性[18]。肠道黏膜受到病原菌的刺激降低,营养物质在肠道中的吸收环境得到改善,进而提高了机体对蛋白质和脂质的利用能力[19]。谷丙转氨酶、谷草转氨酶和碱性磷酸酶的活性是评估肝脏毒性的重要依据,病理表现或毒性都会导致谷草转氨酶和谷丙转氨酶活性升高[20]。此外,谷丙转氨酶、谷草转氨酶和碱性磷酸酶的活性是衡量肝脏损伤的特定指标[21]。在本研究中,饲粮添加AHL酶对21和42日龄肉鸡血清谷丙转氨酶和谷草转氨酶活性无显著影响,表明AHL酶的使用未对肉鸡肝脏造成负面影响。碱性磷酸酶参与脂肪的转化吸收、抵御脂多糖对肠上皮细胞的破坏,也在一定程度上反映机体的生长速度[22]。AHL酶提高了21日龄肉鸡血清中碱性磷酸酶活性,但对42日龄肉鸡血清中碱性磷酸酶活性无显著影响,这与杨伟平等[23]的研究结果一致。

3.2 AHL酶对肉鸡血清免疫指标的影响

肉鸡体内的免疫球蛋白,如IgA、IgM和IgG,在对抗细菌、病毒和毒素,保护免疫系统和维持机体健康方面发挥重要作用[24-25]。IgG由浆细胞分泌,作为抗病毒、抗菌和毒素中和剂[26]。IgA作为抵御病原体入侵的"先驱",已被证明可以改善肠道黏膜的屏障功能,抑制病毒生长[27]。此外,IgM作为与抗原反应的第1个抗体,通过激活补体系统来发挥溶解细菌和病毒等免疫功能[28-29]。在饲粮中添加AHL酶显著提高21和42日龄肉鸡血清中IgA和IgM含量。既往研究表明,AHL酶可以通过干扰QS系统来防止病原体感染宿主[30-31]。饲粮中添加AHL酶可以抑制肠道中有害细菌的生长,并在维持肠道稳态和抑制炎症反应中发挥积极作用,这可能被解释为肉鸡免疫功能改善的原因[32]。研究还发现,AHL酶可以显著提高肉鸡胰腺指数,其中500 mg/kg的添加剂量效果最为显著。

3.3 AHL酶对肉鸡肠道形态的影响

绒毛高度和隐窝深度可用于评估肠道的发育、消化和吸收能力[33-34]。绒毛高度和隐窝深度代表小肠的消化和吸收能力以及上皮细胞的成熟度,而绒隐比代表小肠的消化和吸收功能[35-36]。在本研究中,饲粮添加AHL酶显著提高肉鸡十二指肠、空肠和回肠的绒毛高度和绒隐比,并降低了回肠的隐窝深度。绒毛高度的增加表明肉鸡吸收饲粮中营养物质的消化利用能力得到改善[37],而隐窝深度的降低则表明了绒毛尖端受到的损伤减弱[38]。本试验结果表明,AHL酶显著提高了肉鸡对营养物质的利用率,这也解释了生长性能改善的原因。饲粮中添加AHL酶提高了肉鸡肠道对饲粮的消化利用率,从而显著降低平均日采食量和料重比,因此显著提高蛋白质和能量的利用率。

4 结论

饲粮中添加AHL酶提高了21和42日龄肉鸡血清总蛋白、白蛋白、球蛋白、肌酐和脂蛋白的含量,以及21日龄血清尿素氮含量、碱性磷酸酶活性和42日龄血清总胆固醇含量;饲粮中添加AHL酶提高了21和42日龄肉鸡血清中IgA和IgM含量,以及42日龄的胰腺指数;饲粮中添加AHL酶显著提高肉鸡十二指肠、空肠和回肠的绒毛高度和绒隐比,显著降低了回肠隐窝深度。综合各项指标,AHL酶在肉鸡饲粮中的适宜添加剂量为500 mg/kg。
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