Chinese Journal of Animal Nutrition >

2026 , Vol. 38 >Issue 2: 1048 - 1060

DOI: https://doi.org/10.12418/CJAN2026.084

RESEARCH PAPER

Effects of Compound Probiotics on Growth Performance, Antioxidant Capacity, Immune Function and Intestinal Microbiota of Broilers

  • SHANG Enci , 1 ,
  • LIU Xinhe 1 ,
  • ZU Mengnan 1 ,
  • GAO Fuxian 1 ,
  • LIU Chaoqi 1 ,
  • YIN Qingqiang 1 ,
  • LI Maolong 2 ,
  • DANG Xiaowei 2 ,
  • WANG Lijun , 1, * ,
  • CHANG Juan , 1, *
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  • 1 College of Animal Science and Technology, Henan Agricultural University, Zhengzhou 450046, China
  • 2 Henan Delin Biological Products Co., Ltd., Xinxiang 453000, China
* WANG Lijun, lecturer, E-mail: ;
CHANG Juan, professor, E-mail:

Received date: 2025-07-17

  Online published: 2026-02-12

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Abstract

This experiment was conducted to study the effects of compound probiotics on growth performance, antioxidant capacity, immune function and intestinal microbiota of broilers. A total of 120 one-day-old Arbor Acres broilers were randomly divided into 2 groups, each consisting of 6 replicates with 10 broilers per replicate. The control group was fed a basal diet, and the experimental group was fed the basal diet supplemented with 200 mg/kg compound probiotics (Bacillus subtilis 1.0×1011 CFU/g, Bacillus licheniformis 1.0×1011 CFU/g, Enterococcus faecalis 2.0×1010 CFU/g, and Saccharomyces cerevisiae 2.5×1010 CFU/g). The experiment lasted for 42 days. The results showed that, compared with the control group, the diarrhea rate and mortality rate of broilers in the experimental group were significantly decreased (P<0.05); the activities of superoxide dismutase and glutathione peroxidase in serum were significantly increased (P<0.05), while the malondialdehyde content in both serum and liver was significantly decreased (P<0.05); the immunoglobulin M content in serum was significantly increased (P<0.05), while the contents of interleukin-6 and tumor necrosis factor-α in serum were significantly decreased (P<0.05); the beta diversity of cecal microbiota was significantly different (P<0.05); at the phylum level, the relative abundances of Firmicutes and Cyanobacteria in cecum were significantly increased (P<0.05), while the Bacteroidota relative abundance in cecum was significantly decreased (P<0.05); at the genus level, the relative abundances of Bacteroides, Alistipes and Ruminococcus_torques_group in cecum were significantly decreased (P<0.05), while the relative abundances of Faecalibacterium, Phascolarctobacterium and Lactobacillus in cecum were significantly increased (P<0.05). It is concluded that the dietary supplementation of compound probiotics can reduce the diarrhea rate and mortality rate of broilers, enhance the antioxidant capacity and immune function, and promote the health of animals by regulating intestinal microbiota.

Key words： broilers; compound probiotics; growth performance; antioxidant capacity; immune function; intestinal microbiota

Cite this article

SHANG Enci , LIU Xinhe , ZU Mengnan , GAO Fuxian , LIU Chaoqi , YIN Qingqiang , LI Maolong , DANG Xiaowei , WANG Lijun , CHANG Juan . Effects of Compound Probiotics on Growth Performance, Antioxidant Capacity, Immune Function and Intestinal Microbiota of Broilers[J]. Chinese Journal of Animal Nutrition, 2026 , 38(2) : 1048 -1060 . DOI: 10.12418/CJAN2026.084

我国是畜禽养殖大国,同时也是抗生素使用大国,数据显示,我国畜牧及饲料行业抗生素的用量占据国内消费抗生素总量的50%以上[1]。尽管2020年饲料端全面禁抗显著改善了抗生素的滥用问题,但环境污染、抗生素残留及致病菌耐药性增加等不良后果仍持续存在,阻碍养殖业的发展[2]。在这一背景下,研发安全、高效的抗生素替代品成为动物营养与饲料科学领域的研究热点[3]。
益生菌是一类可定植于动物肠道,且可以促进肠道健康、增强消化吸收功能、提高动物生产性能的微生物统称[4]。在动物养殖中,诸多学者针对益生菌的功能和益生作用进行了研究。马萌等[5]在基础饲粮中添加枯草芽孢杆菌,结果表明促进了肉鸡生长性能的提高,并提高了肉鸡的经济效益。马佰贺等[6]在基础饲粮中添加地衣芽孢杆菌,结果表明促进了肉鸡生长性能的提高,并增加了肠道乳酸菌数量,抑制了有害菌增殖。胥彩玉[7]研究表明,在基础饲粮中添加粪肠球菌对肉鸡生长性能、肉品质和抗氧化指标等具有促进作用。徐杰等[8]在基础饲粮中添加包被活性干酵母对仔猪生长性能和腹泻率具有改善作用。孙海哨等[9]利用400 mg/kg复合益生菌(枯草芽孢杆菌+乳酸菌+产朊假丝酵母菌)饲喂肉鸡,显著提高了肉鸡生长性能,并显著提高了血清免疫球蛋白含量及免疫器官指数。章岳军等[10]在灵山土鸡的研究中发现,复合益生菌对其生长性能未产生显著影响,但可以显著增强免疫功能并改善肠道形态结构。上述研究表明,益生菌及复合益生菌添加剂可以提高肉鸡生长性能、改善腹泻率和免疫功能,但由于益生菌种类、组合方式及添加量等不同,复合益生菌制剂在动物体内产生的效果不同。因此,本试验选择枯草芽孢杆菌、地衣芽孢杆菌、粪肠球菌和酿酒酵母复合制剂添加至基础饲粮中,探讨其对肉鸡生长性能、抗氧化能力、免疫功能和肠道菌群的影响,为肉鸡养殖业的绿色发展提供参考。

1 材料与方法

1.1 试验材料

复合益生菌由枯草芽孢杆菌(活菌数1.0×1011 CFU/g)、地衣芽孢杆菌(活菌数1.0×1011 CFU/g)、粪肠球菌(活菌数2.0×1010 CFU/g)和酿酒酵母(活菌数2.5×1010 CFU/g)组成。

1.2 试验设计

本试验方案获河南农业大学动物福利与伦理委员会批准,批准编号:SKLAB-B-2010-003-01。试验选取1日龄健康、体重相近的雄性爱拔益加(AA)肉鸡120只,随机分为2组,每组6个重复,每个重复10只鸡。对照组饲喂基础饲粮,试验组在饲喂基础饲粮的基础上添加200 mg/kg复合益生菌。试验期42 d,分为1~21日龄和22~42日龄2个阶段。
基础饲粮参照《鸡饲养标准》(NY/T 33—2004)配制,其组成及营养水平见表1。饲粮中粗蛋白质、钙和总磷含量分别参照GB/T 6432—2018[11]、GB/T 6436—2018[12]和GB/T 6437—2018[13]中方法测定,代谢能以及有效磷和氨基酸含量参照《中国饲料成分及营养价值表(2020年第31版)》[14]计算。
表1 基础饲粮组成及营养水平(风干基础)

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

项目
Items		 含量Content
1~21日龄
1 to 21
days of age		 22~42日龄
22 to 42
days of age
原料Ingredients
玉米Corn 59.13 60.77
豆粕Soybean meal 35.15 32.00
大豆油Soybean oil 1.00 3.00
磷酸氢钙CaHPO4 1.66 1.53
石粉Limestone 1.40 1.23
DL-蛋氨酸DL-Met 0.20 0.10
L-赖氨酸L-Lys 0.09
食盐NaCl 0.37 0.37
预混料Premix1) 1.00 1.00
合计Total 100.00 100.00
营养水平Nutrient levels2)
代谢能ME/(MJ/kg) 12.34 12.91
粗蛋白质CP 20.70 19.30
钙Ca 1.00 0.90
总磷TP 0.68 0.64
有效磷AP 0.43 0.41
赖氨酸Lys 1.15 1.00
蛋氨酸Met 0.52 0.41
蛋氨酸+胱氨酸Met+Cys 0.88 0.75

1)预混料为每千克饲粮提供 The premix provided the following per kg of diets:VA 12 000 IU,VD3 4 350 IU,VE 3 IU,VK3 3.9 mg,VB1 3 mg,VB2 7.5 mg,VB6 4.2 mg,VB12 0.3 mg,D-生物素 D-biotin 0.15 mg,叶酸 folic acid 1.2 mg,烟酰胺 nicotinamide 33 mg,D-泛酸 D-pantothenic acid 12 mg,Cu (as basic copper chloride) 5.1 mg,Fe (as ferrous sulfate) 72 mg,Mn (as manganese sulfate) 83 mg,Zn (as zinc sulfate) 56 mg,I (as calcium iodate) 0.56 mg,Se (as sodium selenite) 0.21 mg。

2)粗蛋白质、钙和总磷含量为测定值,其余为计算值。CP, Ca and TP were determined values, while the others were calculated values.

1.3 饲养管理

动物饲养试验在河南新乡河南农业大学原阳试验基地进行,采用多层立体笼养,自由采食和饮水,舍内适当通风。人工光照制度为:7日龄内24 h光照;7日龄后1 h黑暗,23 h光照。免疫程序为:7日龄免疫新支H120二连弱毒苗,14日龄免疫法氏囊弱毒苗,20日龄免疫新城疫C30弱毒苗。

1.4 测定指标及方法

1.4.1 生长性能

在整个饲养周期内准确称量并记录肉鸡的初始体重以及22和43日龄空腹体重,记录每天的采食量并回收抛洒料,同时观察试验鸡的健康状况,用于计算平均体重、平均日增重(ADG)、平均日采食量(ADFI)、料重比(F/G)、死亡率(mortality rate,MR)和腹泻率(diarrhea rate,DR)。

1.4.2 血清和肝脏抗氧化指标

试验结束后(43日龄),每重复选取1只试验鸡进行翅静脉采血,离心后取上清液分装,置于-20 ℃冰箱保存备用;试验鸡采血后屠宰,取肝脏组织样品,液氮速冻后-80 ℃保存备用。
血清和肝脏抗氧化指标采用试剂盒(南京建成生物工程研究所)进行测定,测定指标包括: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)、谷胱甘肽-S-转移酶(glutathione-S-transferase,GST)活性以及丙二醛(malondialdehyde,MDA)含量和总抗氧化能力(total antioxidant capacity,T-AOC)。

1.4.3 血清免疫指标

血清免疫指标采用酶联免疫吸附试验(ELISA)法测定,试剂盒由上海优选生物科技有限公司提供,测定指标包括:免疫球蛋白A(IgA)、免疫球蛋白G(IgG)、免疫球蛋白M(IgM)、白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)含量。

1.4.4 盲肠微生物区系

试验鸡屠宰后,取盲肠内容物,经处理后委托上海美吉生物医药科技有限公司进行测序分析,测序步骤及后续如下:1)样品DNA的抽提。根据E.Z.N.A.® Soil DNA Kit(Omega Bio-Tek,美国)说明书进行抽提。2)PCR扩增。利用引物对338F(5'-ACTCCTACGGGAGGCAGCAG-3')和806R(5'-GGACTACHVGGGTWTCTAAT-3')[15]扩增16S rRNA的V3~V4可变区。3)MiSeq文库构建、质检及测序。采用NEXTFLEX® Rapid DNA-Seq Kit对纯化后的PCR产物进行建库。4)生物信息学分析,所有分析均在美吉云平台上进行。alpha多样性采用mothur软件进行分析[16],alpha多样性的组间差异分析采用Wilcoxon秩和检验;beta多样性采用主坐标分析(PCoA)(基于Bray-Curtis距离)判定样本间的相似性,并通过非参数检验置换多元方差分析(PERMANOVA)评估样本间微生物群落结构的差异显著性;采用线性判别分析(LDA)效应大小(LEfSe)确定不同组间微生物从门到属水平相对丰度差异显著的类群[17]。5)测序后将原始数据上传至NCBI SRA数据库(登记号:PRJNA1293786)。

1.5 数据统计和分析

试验数据经Excel 2024初步整理后,采用SPSS 26.0软件对各组数据进行独立样本t检验,P<0.05表示差异显著,结果数据采用“平均值±标准差”形式表示。

2 结果与分析

2.1 复合益生菌对肉鸡生长性能的影响

由表2可知,在饲养前期(1~21日龄),与对照组相比,试验组肉鸡腹泻率和死亡率显著降低(P<0.05);在饲养后期(22~42日龄),与对照组相比,试验组腹泻率显著降低(P<0.05),且终末体重和ADG有所提高(P>0.05);在整个试验期(1~42日龄),与对照组相比,试验组腹泻率和死亡率显著降低(P<0.05),且终末体重和ADG有所提高(P>0.05)。上述结果表明,饲粮添加复合益生菌具有降低肉鸡腹泻率和死亡率的作用。
表2 复合益生菌对肉鸡生长性能的影响

Table 2 Effects of compound probiotics on growth performance of broilers

项目
Items		 初始体重
Initial
BW/g		 终末体重
Final
BW/g		 平均日采食量
ADFI/
(g/d)		 平均日增重
ADG
(g/d)		 料重比
F/G		 腹泻率
DR/%		 死亡率
MR/%
1~21日龄1 to 21 days of age
对照组
Control group		 44.68
±0.48		 626.28
±13.79		 36.94
±0.94		 27.70
±0.66		 1.33
±0.03		 9.37
±1.15a		 0.40
±0.19a
试验组
Experimental group		 44.75
±0.46		 615.67
±18.50		 36.44
±1.12		 27.19
±0.88		 1.34
±0.02		 6.75
±0.36b		 0.08
±0.19b
P值P-value 0.811 0.286 0.418 0.281 0.621 0.018 <0.001
22~42日龄22 to 42 days of age
对照组
Control group		 626.28
±13.79		 1 814.43
±66.06		 103.81
±3.94		 56.58
±2.98		 1.84
±0.07		 3.41
±0.47a		 0.00
±0.00
试验组
Experimental group		 615.67
±18.50		 1 824.87
±44.61		 107.05
±7.16		 57.58
±1.71		 1.86
±0.08		 1.82
±0.36b		 0.00
±0.00
P值P-value 0.286 0.755 0.354 0.491 0.654 <0.001 1.000
1~42日龄1 to 42 days of age
对照组
Control group		 44.68
±0.48		 1 814.43
±66.06		 70.37
±2.21		 42.14
±1.56		 1.67
±0.04		 6.39
±0.65a		 0.20
±0.10a
试验组
Experimental group		 44.75
±0.46		 1 824.87
±44.61		 71.74
±4.01		 42.38
±1.06		 1.69
±0.06		 4.29
±0.21b		 0.04
±0.10b
P值P-value 0.811 0.755 0.756 0.485 0.484 0.018 <0.001

同列数据肩标不同字母表示差异显著(P<0.05),无字母表示差异不显著(P>0.05)。下表同。

In the same column, values with different letter superscripts mean significant differences (P<0.05), while with no letter superscripts mean no significant differences (P>0.05). The same as below.

2.2 复合益生菌对肉鸡血清和肝脏抗氧化指标的影响

由表3可知,与对照组相比,试验组肉鸡血清MDA含量显著降低(P<0.05),血清SOD和GSH-Px活性显著提高(P<0.05)。由表4可知,与对照组相比,试验组肉鸡肝脏MDA含量显著降低(P<0.05),肝脏其他抗氧化指标无显著差异(P>0.05)。以上结果表明,饲粮添加复合益生菌可改善肉鸡机体抗氧化能力。
表3 复合益生菌对肉鸡血清抗氧化指标的影响

Table 3 Effects of compound probiotics on serum antioxidant indices of broilers

项目
Items		 丙二醛
MDA/(nmol/mL)		 过氧化氢酶
CAT/(U/mL)		 总抗氧化能力
T-AOC/(U/mL)		 超氧化物歧化酶
SOD/(U/mL)		 谷胱甘肽过氧化物酶
GSH-Px/(U/mL)
对照组Control group 1.94±0.07a 7.38±1.34 0.94±0.12 17.88±3.01b 1 706.25±59.39b
试验组Experimental group 1.71±0.18b 6.84±1.06 1.19±0.32 44.59±3.84a 1 965.00±99.27a
P值P-value 0.012 0.455 0.111 <0.001 <0.001
表4 复合益生菌对肉鸡肝脏抗氧化指标的影响

Table 4 Effects of compound probiotics on liver antioxidant indices of broilers

项目
Items		 丙二醛
MDA/
(nmol/mg prot)		 过氧化氢酶
CAT/
(U/mg prot)		 总抗氧化能力
T-AOC/
(U/mg prot)		 超氧化物
歧化酶
SOD/
(U/mg prot)		 谷胱甘肽
过氧化物酶
GSH-Px/
(U/mg prot)		 谷胱甘肽-
S-转移酶
GST/
(U/mg prot)
对照组Control group 7.22±1.00a 0.38±0.04 1.28±0.20 21.10±4.07 248.23±57.32 348.73±42.56
试验组Experimental group 3.64±0.80b 0.38±0.06 1.22±0.21 22.33±2.73 241.06±43.68 342.88±30.30
P值P-value <0.001 0.913 0.571 0.553 0.812 0.789

2.3 复合益生菌对肉鸡血清免疫指标的影响

由表5可知,与对照组相比,试验组肉鸡血清IL-6和TNF-α含量显著降低(P<0.05),血清IgM含量显著提高(P<0.05),表明饲粮添加复合益生菌可在一定程度上增强肉鸡免疫功能。
表5 复合益生菌对肉鸡血清免疫指标的影响

Table 5 Effects of compound probiotics on serum immune indices of broilers

项目
Items		 白细胞
介素-6
IL-6/
(pg/mL)		 白细胞
介素-1β
IL-1β/
(pg/mL)		 肿瘤坏死
因子-α
TNF-α/
(pg/mL)		 免疫球
蛋白A
IgA/
(μg/mL)		 免疫球
蛋白G
IgG/
(g/mL)		 免疫球
蛋白M
IgM/
(μg/mL)
对照组Control group 36.13±3.48a 179.78±9.29 116.38±8.19a 1 685.39±144.97 30.12±0.34 2 351.25±134.65b
试验组
Experimental group		 30.86±1.14b 167.60±14.61 81.33±2.38b 1 743.37±32.35 30.86±2.20 2 643.23±81.27a
P值P-value 0.028 0.209 <0.001 0.465 0.552 0.010

2.4 复合益生菌对肉鸡盲肠微生物区系的影响

2.4.1 复合益生菌对肉鸡盲肠微生物群落alpha和beta多样性的影响

如图1所示,与对照组相比,试验组肉鸡盲肠微生物群落Ace指数、Chao指数、Shannon指数和Simpson指数均无显著差异(P>0.05),表明饲粮添加复合益生菌对肉鸡盲肠微生物群落alpha多样性无显著影响。
图1 肉鸡盲肠微生物群落alpha多样性分析

Fig.1 Alpha diversity analysis of cecal microbiota of broilers

如图2所示,对照组与试验组间样品较为分离,肉鸡盲肠微生物群落beta多样性存在显著差异(P<0.05),表明饲粮添加复合益生菌可改变肉鸡盲肠菌群结构。
图2 肉鸡盲肠微生物群落beta多样性分析

Fig.2 Beta diversity analysis of cecal microbiota of broilers

2.4.2 复合益生菌对肉鸡盲肠微生物群落在门水平上相对丰度的影响

如图3-A所示,在门水平上,肉鸡盲肠菌群主要由厚壁菌门(Firmicutes)和拟杆菌门(Bacteroidota)组成,这2种菌门相对丰度占总菌门的97%以上,为优势菌群;其次是放线菌门(Actinobacteriota)、蓝细菌门(Cyanobacteria)、变形菌门(Proteobacteria)、脱硫杆菌门(Desulfobacterota)和弯曲杆菌门(Campylobacterota)等。如图3-B所示,与对照组相比,试验组肉鸡盲肠厚壁菌门和蓝细菌门相对丰度显著提高(P<0.05),盲肠拟杆菌门相对丰度显著降低(P<0.05),盲肠放线菌门无显著差异(P>0.05)。
图3 肉鸡盲肠微生物群落在门水平上相对丰度

数据柱形标注“*”表示差异显著(P<0.05),“ns”表示差异不显著(P>0.05)。图4同。

Fig.3 Relative abundance of cecal microbiota of broilers at phylum level

Value columns with “*” mean significant differences (P<0.05), and “ns” mean no significant differences (P>0.05). The same as Fig.4.

2.4.3 复合益生菌对肉鸡盲肠微生物群落在属水平上相对丰度的影响

如图4-A所示,肉鸡盲肠菌群在属水平上主要包括另枝菌属(Alistipes)、拟杆菌属(Bacteroides)、梭菌纲未定级目UCG-014属(norank_o_Clostridia_UCG-014)、粪杆菌属(Faecalibacterium)、考拉杆菌属(Phascolarctobacterium)、瘤胃球菌属扭链群(Ruminococcus_torques_group)和乳杆菌属(Lactobacillus)等10余种菌属,其中另枝菌属和拟杆菌属为优势菌属。
图4 肉鸡盲肠微生物群落在属水平上相对丰度

Fig.4 Relative abundances of cecal microbiota of broilers at genus level

如图4-B所示,与对照组相比,试验组肉鸡盲肠拟杆菌属、另枝菌属和瘤胃球菌属扭链群相对丰度显著降低(P<0.05),盲肠粪杆菌属、考拉杆菌属和乳杆菌属相对丰度显著提高(P<0.05)。

3 讨论

3.1 复合益生菌对肉鸡生长性能的影响

生长性能是衡量肉鸡生产效益的关键指标,主要包括ADG、ADFI和F/G等指标[18]。已有研究表明,在饲粮中添加益生菌可以降低肉鸡腹泻率[19]。吕紫一等[20]研究发现,复合益生菌能显著改善雏鸡生长性能和小肠组织形态,减缓大肠杆菌对肠道造成的损伤,进而减少腹泻发生。此外,枯草芽孢杆菌等产酶型菌株分泌的淀粉酶、蛋白酶等与机体协同作用,能使更多的养分为动物机体所吸收,进而促进生长[21]。张煜等[22]在饲粮中添加0.30%复合益生菌制剂可改善肉鸡的生长性能和肠道健康,增强免疫力。不过,复合益生菌对生长性能的影响存在差异性,部分研究显示其具有促进作用[23-24],而另外一些研究则表明其作用有限[6,25],此差异可能源于菌种特性及协同增效作用不同。本试验结果表明,饲粮添加复合益生菌后,整个饲养周期内肉鸡腹泻率和死亡率均显著降低,终末体重和ADG有所提高,表明复合益生菌对维持肉鸡健康具有促进作用。

3.2 复合益生菌对肉鸡抗氧化能力的影响

动物体内抗氧化酶主要包括SOD、GSH-Px和CAT等,这些抗氧化酶通过不同的机制发挥协同作用,并通过清除体内的活性氧(ROS)来维持机体内氧化与抗氧化的动态平衡,从而保护机体免受氧化损伤[26]。MDA是脂质过氧化的次级代谢物,其含量直接反映细胞膜中ROS攻击的程度,与细胞损伤密切相关,被认为是检测脂质过氧化程度的指标[27]。安楠[28]在饲粮中添加复合菌显著提高了肉牛血清SOD和GSH-Px活性,显著降低了血清MDA含量。金晶等[29]在饲粮中添加0.3%复合益生菌可提高肉鸭血清和肠道抗氧化能力,促进肉鸭生长发育。樊华等[30]研究发现,饲粮添加1%和2%复合益生菌可显著降低肉牛血清MDA含量,显著提高血清GSH-Px活性。鲁玲君[31]研究表明,小鼠补充复合益生菌能抑制脂质过氧化标志物MDA的产生并提高谷胱甘肽含量。本试验结果表明,饲粮添加复合益生菌后肉鸡血清MDA含量显著降低,血清SOD和GSH-Px活性显著提高,同时肝脏MDA含量显著降低,说明复合益生菌能够有效抑制脂质过氧化反应,保护细胞膜结构和功能的完整性,提高肉鸡抗氧化能力。

3.3 复合益生菌对肉鸡免疫功能的影响

炎症因子(如IL-6、IL-1β和TNF-α)参与细胞凋亡和焦亡等多种细胞死亡途径,可促进细胞死亡和组织损伤[32]。IL-6能够刺激白细胞增殖、抗体产生和调节炎症反应;TNF-α可刺激炎症介质的产生,导致炎症反应加剧[33]。本试验结果表明,饲粮添加复合益生菌后,肉鸡血清促炎因子IL-6和TNF-α含量显著降低。宋丹等[34]在AA肉公鸡饲粮中添加1 g/kg包被植物乳杆菌和1 g/kg屎肠球菌可提高血清抗炎因子白细胞介素-10(IL-10)含量,降低血清促炎因子TNF-α、白细胞介素-2(IL-2)和IL-6含量。此外,益生菌可有效调节动物机体的免疫系统,通过平衡机体内的抗炎因子与促炎因子含量来激活机体的免疫系统,防止病原微生物的侵害,稳定动物肠道的内环境[35]。
免疫球蛋白IgA、IgG、IgM与动物机体的免疫反应密切相关,是评估机体免疫能力的重要指标,也是抵抗感染的重要物质[2]。IgG是血液中占比最大且具有抵抗感染能力的一类免疫球蛋白,而IgM是高效的免疫活性物质[36]。Bai等[37]报道,在肉鸡饲粮中添加枯草芽孢杆菌可以提高其血清IgG和IgA含量。王琳等[38]研究显示,枯草芽孢杆菌对肉鸡血清IgA含量无显著影响,但可显著提高其血清IgM含量。这与本试验结果一致,本试验中,饲粮添加复合益生菌后,肉鸡血清IgM含量显著升高,说明枯草芽孢杆菌、地衣芽孢杆菌、粪肠球菌和酿酒酵母组成的复合益生菌能一定程度上增强肉鸡免疫功能,促进其肠道健康。这种现象的潜在机制可能与复合益生菌介导的免疫调节功能相关。研究表明,饲粮添加复合益生菌可以显著提高肉鸡生长性能和免疫功能[39]。张彩凤[40]研究表明,饲粮添加酵母菌制剂显著提高肉鸡抗氧化能力和免疫功能。连丽娜等[41]研究表明,枯草芽孢杆菌显著提高黄羽肉鸡免疫力,并改善生长性能。由此可知,本试验所用复合益生菌具有增强肉鸡免疫功能和降低炎症反应的作用。

3.4 复合益生菌对肉鸡肠道菌群的影响

动物胃肠道菌群由益生菌、病原菌和共生菌共同组成,在门水平上又主要分为厚壁菌门、拟杆菌门和蓝细菌门等。其中,厚壁菌门对于肠道健康、免疫调节和整体健康都具有正面影响,研究发现厚壁菌门可通过在肠道中释放细胞壁糖缀合物提高机体抵抗病原微生物的能力[42]。冯宇妍[43]研究表明,复合益生菌显著提高肉鸡肠道厚壁菌门相对丰度,改善肠道健康。研究报道,拟杆菌门参与多种微生物疾病的发生,在免疫系统受损或肠道微生态失衡的情况下可引起由感染导致的疾病发生[44]。蓝细菌门是一种具有抗炎、抗氧化和伤口愈合特性的微生物,是抗菌肽的潜在来源[45]。本试验中,饲粮添加由枯草芽孢杆菌、地衣芽孢杆菌、粪肠球菌和酿酒酵母菌组成的复合益生菌使肉鸡盲肠厚壁菌门和蓝细菌门相对丰度显著提高,盲肠拟杆菌门相对丰度显著降低,表明复合益生菌可以降低肉鸡疾病的发生进而起到改善动物健康的作用。
在属水平上,2组肉鸡盲肠微生物群落以另枝菌属和拟杆菌属为优势菌属。刘韬虞[46]研究表明,拟杆菌属参与多种微生物疾病的发生,在肠道微生态失衡的情况下引起肠道感染致病菌,进而导致疾病的发生。另枝菌属主要存在于健康的肠道中,但也可从阑尾炎患者腹部和脑脓肿中分离出来,这一现象表明另枝菌属有潜在的致病风险[47]。研究报道,乳杆菌属和粪杆菌属是常见的益生菌[48],其中乳杆菌属的大多数菌株已被用作益生菌[49]。乳杆菌属还是天然抗氧化剂的替代品,具有增强机体免疫力和抗氧化等功效,能够减少宿主自由基氧化应激造成的损害[50]。粪杆菌属是肠道内丰富的产丁酸菌,其分解粗纤维产生的丁酸可以作为肠上皮细胞的能量来源,也可以调节肠道pH以抑制有害菌的繁殖;此外,粪杆菌属具有良好的抗炎特性,有助于炎症性肠病紧密连接屏障的恢复[51]。考拉杆菌属属于厚壁菌门,可以在肠道内发酵产生短链脂肪酸,短链脂肪酸可以为肠上皮细胞提供能量,降低动物体内炎症水平,并增强消化道消化吸收能力[52]。考拉杆菌属还通过与病原菌竞争营养物质来有效抑制病原菌在肠道中的生长与定植[53]。瘤胃球菌属扭链群通常存在于肠道中,是一种“有害细菌”,其增加可能会损坏肠道屏障,同时也与胃肠道疾病相关[54]。本试验中,饲粮添加复合益生菌后,肉鸡盲肠拟杆菌属、另枝菌属和瘤胃球菌属扭链群相对丰度显著降低,盲肠粪杆菌属、考拉杆菌属和乳杆菌属相对丰度显著提高,表明复合益生菌具有调节肠道微生态平衡,维持肠道健康的作用。

4 结论

饲粮添加200 mg/kg复合益生菌能够降低肉鸡腹泻率和死亡率,增强抗氧化能力和免疫功能,并通过调节肠道菌群改善机体健康。

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