动物营养学报 >

2024 , Vol. 36 >Issue 5: 3024 - 3034

DOI: https://doi.org/10.12418/CJAN2024.260

研究论文

饲粮添加糖萜素和植物乳杆菌对肉鹅免疫功能、肠道形态和肠道菌群结构的影响

  • 钱旺 , 1 ,
  • 王宝维 , 1, 2, * ,
  • 张名爱 1 ,
  • 凡文磊 1 ,
  • 孔敏 1 ,
  • 代林 2 ,
  • 谭绍兴 1 ,
  • 王秉翰 3
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  • 1 青岛农业大学动物科技学院,青岛 266109
  • 2 青岛农业大学食品科学与工程学院,青岛 266109
  • 3 青岛慧和生物科技有限公司,青岛 266109
* 王宝维,教授,硕士生导师,E-mail:

钱 旺(1997—),男,河北定州人,硕士研究生,研究方向为动物营养与保健。E-mail:

Office editor: 武海龙

收稿日期: 2023-11-30

  网络出版日期: 2024-05-15

基金资助

2021年度山东省重点研发计划(2021CXGC010805)

财政部和农业农村部-国家现代农业产业技术体系资助(CARS-42-14)

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Effects of Dietary Supplementation of Saccharicter-Penin and Lactobacillus plantarum on Immune Function, Intestinal Morphology and Intestinal Flora Structure of Meat Geese

  • QIAN Wang , 1 ,
  • WANG Baowei , 1, 2, * ,
  • ZHANG Ming’ai 1 ,
  • FAN Wenlei 1 ,
  • KONG Min 1 ,
  • DAI Lin 2 ,
  • TAN Shaoxing 1 ,
  • WANG Binghan 3
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  • 1 College of Animal Science and Technology, Qingdao Agricultural University, Qingdao 266109, China
  • 2 College of Food Science and Engineering, Qingdao Agricultural University, Qingdao 266109, China
  • 3 Qingdao Huihe Biotechnology Co., Ltd., Qingdao 266109, China
* professor, E-mail:

Received date: 2023-11-30

  Online published: 2024-05-15

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摘要

本试验旨在研究饲粮添加糖萜素和植物乳杆菌对肉鹅免疫功能、肠道形态和肠道菌群结构的影响,以探讨糖萜素和植物乳杆菌协同关系与功能。选取96只体重为(0.135±0.020) kg的1日龄五龙鹅,随机分成4组,每组4个重复,每重复6只鹅。对照组(CON组)饲喂基础饲粮,糖萜素组(SP组)饲喂基础饲粮+0.1%糖萜素,植物乳杆菌组(LP组)饲喂基础饲粮+0.4%植物乳杆菌,糖萜素+植物乳杆菌组(SP+LP组)饲喂基础饲粮+0.1%糖萜素+0.4%植物乳杆菌。试验期70 d。结果表明:1)与CON组相比,SP组的血清免疫球蛋白M(IgM)含量显著提高(P<0.05),LP组的血清免疫球蛋白G(IgG)含量显著提高(P<0.05),SP+LP组的血清IgM和干扰素-γ(IFN-γ)含量显著提高(P<0.05)。2)与CON组相比,SP组、LP组和SP+LP组的空肠白细胞介素-1β(IL-1β)、白细胞介素-2(IL-2)和肿瘤坏死因子-α(TNF-α)含量显著降低(P<0.05),空肠白细胞介素-4(IL-4)和分泌型免疫球蛋白A(sIgA)含量显著提高(P<0.05)。3)与CON组相比,SP组、LP组和SP+LP组的空肠绒毛高度显著提高(P<0.05);LP组和SP+LP组的空肠隐窝深度显著降低(P<0.05),空肠绒毛高度/隐窝深度显著提高(P<0.05)。与CON组相比,LP组和SP+LP组的回肠绒毛高度显著提高(P<0.05);SP组、LP组和SP+LP组的回肠隐窝深度显著降低(P<0.05),回肠绒毛高度/隐窝深度显著提高(P<0.05)。4)在门水平上,与对照组相比,LP组和SP+LP组的盲肠拟杆菌门相对丰度显著提高(P<0.05),SP组、LP组和SP+LP组的盲肠弯曲菌门相对丰度显著降低(P<0.05)。在属水平上,与对照组相比,LP组的盲肠拟杆菌属相对丰度显著提高(P<0.05),LP组和SP+LP组的盲肠巴恩斯氏菌属相对丰度显著提高(P<0.05),SP+LP组的盲肠普氏栖粪杆菌属相对丰度显著提高(P<0.05)。综上所述,饲粮中联合添加糖萜素和植物乳杆菌可以增强五龙鹅的免疫功能,促进肠道形态发育,改善肠道优势菌群结构。

关键词： 五龙鹅; 糖萜素; 植物乳杆菌; 免疫; 细胞因子; 肠道形态; 肠道菌群

本文引用格式

钱旺 , 王宝维 , 张名爱 , 凡文磊 , 孔敏 , 代林 , 谭绍兴 , 王秉翰 . 饲粮添加糖萜素和植物乳杆菌对肉鹅免疫功能、肠道形态和肠道菌群结构的影响[J]. 动物营养学报, 2024 , 36(5) : 3024 -3034 . DOI: 10.12418/CJAN2024.260

Abstract

This experiment was conducted to study the effects of dietary supplementation of saccharicter-penin and Lactobacillus plantarum on immune function, intestinal morphology and intestinal flora structure of meat geese, to explore the synergistic relationship and function of saccharicter-penin and Lactobacillus plantarum. Ninety-six 1-day-old Wulong geese with body weight of (0.135±0.020) kg were randomly divided into 4 groups with 4 replicates per group and 6 geese per replicate. The control group (CON group) was fed a basal diet, the saccharicter-penin group (SP group) was fed the basal diet+0.1% saccharicter-penin, the Lactobacillus plantarum group (LP group) was fed the basal diet+0.4% Lactobacillus plantarum, and the saccharicter-penin+Lactobacillus plantarum group (SP+LP group) fed the basal diet+0.1% saccharicter-penin+0.4% Lactobacillus plantarum. The experiment lasted for 70 days. The results showed as follows: 1) compared to the control group, the serum immunoglobulin M (IgM) content of SP group was significantly increased (P<0.05), the serum immunoglobulin G (IgG) content of LP group was significantly increased (P<0.05), and the contents IgM and interferon-γ (IFN-γ) in serum of SP+LP group were significantly increased (P<0.05). 2) Compared to the control group, the contents of nterleukin-1β (IL-1β), interleukin-2 (IL-2) and tumor necrosis factor-α (TNF-α) in jejunum of SP group, LP group and SP+LP group were significantly decreased (P<0.05), and the contents of interleukin-4 (IL-4) and secretory immunoglobulin A (sIgA) were significantly increased (P<0.05). 3) Compared to the control group, the jejunal villus height of SP group, LP group and SP+LP group was significantly increased (P<0.05); the jejunal crypt depth of LP group and SP+LP group was significantly decreased (P<0.05), and the jejunal villus height/crypt depth was significantly increased (P<0.05). Compared to the control group, the ileal villus height of LP group and SP+LP group was significantly increased (P<0.05); the ileal crypt depth of SP group, LP group and SP+LP group was significantly decreased (P<0.05), and the ileal villus height/crypt depth was significantly increased (P<0.05). 4) At the phylum level, compared to the control group, the cecal Bacteroidota relative abundance of LP group and SP+LP group was significantly decreased (P<0.05), and the cecal Campilobacterota relative abundance of SP group, LP group and SP+LP group was significantly decreased (P<0.05). At the genus level, compared to the control group, the cecal Bacteroides relative abundance of LP group was significantly increased (P<0.05), the cecal Barnesiella relative abundance of LP group and SP+LP group was significantly increased (P<0.05), and the cecal Faecalibacterium relative abundance of SP+LP group was significantly increased (P<0.05). In conclusion, dietary combined supplementation of saccharicter-penin and Lactobacillus plantarum can enhance the immune function of Wulong geese, promote the intestinal morphological development, and improve the structure of intestinal dominant flora.

Key words： Wulong geese; saccharicter-penin; Lactobacillus plantarum; immune; cytokines; intestinal morphology; intestinal flora

随着全国肉类消费量不断增加,禽类生产也迅速增加[1]。众所周知,家禽生产中抗生素的使用可以提高家禽生长性能和抗感染性,然而,随着抗生素的广泛使用,出现一系列的负面影响[2],如造成抗生素的耐药性和药物残留[3],严重危及禽产品的安全性。寻找替代抗生素产品已成为解决这一问题的紧迫需求[4]。植物乳杆菌已经被证实为抗生素的潜在替代品,它有助于刺激生长性能、改善肠道组织形态、提高免疫功能并改善畜禽肉品质[5⇓⇓-8]。糖萜素是山茶科植物中提取的含有皂苷、多糖和有机酸的天然糖甙化合物,也已经被批准为饲料添加剂[8-9]。Chen等[10]研究表明,三萜皂苷有助于增强肠道有益菌、减少硫酸盐还原菌,可调节肠道炎症及改善肠道微生物环境。糖萜素和植物乳杆菌都具有改善畜禽生长性能[11-12]、提高机体免疫功能[13]、调节肠道微生物组成[14-15]、改善肠道形态[16- 17]等效用。因此,饲粮中糖萜素与植物乳杆菌联合添加可能会对肉鹅的健康和生长产生积极的协同影响。然而,目前尚未见关于糖萜素和植物乳杆菌联合添加对肉鹅免疫功能和肠道发育的影响研究。因此,本试验旨在研究饲粮添加糖萜素和植物乳杆菌对肉鹅免疫功能、肠道发育和肠道菌群结构的影响,并研究二者的协同作用,以期为糖萜素和植物乳杆菌在肉禽生产中应用提供理论依据。

1 材料与方法

1.1 试验时间与地点

饲养试验在青岛农业大学国家水禽研究所山东高密水禽试验基地进行。

1.2 试验材料

试验鹅由高密市银河润雁鹅业有限公司提供,品种为五龙鹅(灰羽);糖萜素由山东鑫谷健康产业有限公司提供,纯度≥50%;植物乳杆菌由中国微生物菌种保藏管理委员会普通物生物中心提供,活菌数1×109 CFU/mL。

1.3 试验设计

选取1日龄、体重(0.135±0.020) kg、健康的96只五龙鹅(公母各占1/2),随机分成4组,每组4个重复,每个重复6只鹅。对照组(CON组)饲喂基础饲粮,糖萜素组(SP组)饲喂基础饲粮+0.1%糖萜素,植物乳杆菌组(LP组)饲喂基础饲粮+0.4%植物乳杆菌,糖萜素+植物乳杆菌组(SP+LP组)饲喂基础饲粮+0.1%糖萜素+0.4%植物乳杆菌。糖萜素和植物乳杆菌添加量参照许振华等[18]和Wang等[19]试验结果。基础饲粮组成及营养水平见表1。
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
1~28日龄
1 to 28
days of age		 29~70日龄
29 to 70
days of age
原料Ingredients
玉米Corn 59.68 58.58
豆粕Soybean meal 23.10 20.60
次粉Wheat middling 5.00 6.00
稻壳粉Rice husk meal 1.16 3.50
小麦麸Wheat bran 8.00 8.00
磷酸氢钙CaHPO4 1.40 1.65
石粉Limestone 0.89 0.85
食盐NaCl 0.35 0.40
氯化胆碱Choline chloride 0.07 0.07
预混料Premix1) 0.35 0.35
合计Total 100.00 100.00
营养水平Nutrient levels2)
代谢能ME/(MJ/kg) 12.34 12.12
粗蛋白质CP 17.11 16.05
粗纤维CF 4.54 5.60

1)预混料为每千克饲粮提供 The premix provided the following per kg of diets:VA 9 000 IU,VB1 2.0 mg,VB2 4.0 mg,VB5 40 mg,VB6 4.0 mg,VB12 12 μg,VD3 2 000 IU,VE 40 mg,VK3 0.8 mg,生物素 biotin 0.2 mg,叶酸 folic acid 0.5 mg,D-泛酸 D-pantothenic acid 11 mg,烟酸 nicotinic acid 10 mg,Cu (as copper sulfate) 4 mg,Fe (as ferrous sulfate) 85 mg,Mn (as manganese sulfate) 30 mg,Zn (as zinc sulfate) 65 mg,I (as potassium iodide) 0.30 mg,Se (as sodium selenite) 0.50 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.

1.4 饲养管理

肉鹅采用舍饲笼养,自由采食和饮水,保持自然光照。试验期70 d。

1.5 样品采集与指标测定

1.5.1 饲粮营养水平

饲粮粗蛋白质含量参照GB/T 6432—2018[20]进行测定,粗纤维含量参照GB/T 6234—2006[21]进行测定。代谢能参照《中国饲料成分及营养价值表》(2022年第33版)计算。

1.5.2 血清免疫指标

饲养试验结束后,每个重复随机选取2只肉鹅,翅静脉采血10 mL,3 500 r/min离心10 min分离血清,并于-40 ℃保存,用于检测血清免疫球蛋白G(IgG)、免疫球蛋白M(IgM)、干扰素-γ(IFN-γ)含量。使用南京建成生物工程研究所的试剂盒进行测定。

1.5.3 空肠细胞因子含量

取肉鹅空肠黏膜0.1 g,加入0.9 mL生理盐水破碎匀浆,4 ℃条件下8 000 r/min离心20 min,收集上清液,采用酶联免疫吸附试验(ELISA)试剂盒测定空肠白细胞介素-1β(IL-1β)、白细胞介素-2(IL-2)、白细胞介素-4(IL-4)、肿瘤坏死因子-α(TNF-α)和分泌型免疫球蛋白A(sIgA)含量。

1.5.4 肠道形态

采集肉鹅空肠和回肠组织,于4%多聚甲醛中固定。用乙醇将肠道组织逐步脱水、包埋后切片、苏木素-伊红染色,封片后用显微镜观察并记录小肠绒毛长度和隐窝深度,并计算绒毛高度/隐窝深度。

1.5.5 盲肠菌群结构

通过微生物高通量测序及分析(全场扩增子测序分析)对盲肠内容物进行16S rRNA测序。提取盲肠内容物中的DNA,使用引物338F(5'-ACTCCTACGGGAGGCAGCAG-3')和806R(5'-GGACTACHVGGGTWTCATAAT-3')对16S rRNA V3~V4进行PCR扩增,利用Illumina MiSeq/NovaSeq 6000平台测序,PCR产物回收纯化后,制备文库进行测序分析,并通过群落组成分析并计算α多样性,比较盲肠菌群结构。

1.6 数据处理与统计分析

试验数据用Excel软件进行整理,采用SPSS 26.0统计软件进行单因素方差分析(one-way ANOVA),并通过LSD法进行差异比较,所有数据以平均值和均值标准误(SEM)表示,P<0.05表示差异显著。

2 结果

2.1 饲粮添加糖萜素和植物乳杆菌对肉鹅血清免疫指标的影响

由表2可知,与CON组相比,SP组的血清IgM含量显著提高(P<0.05),LP组的血清IgG含量显著提高(P<0.05),SP+LP组的血清IgM和IFN-γ含量显著提高(P<0.05)。此外,SP+LP组的血清IgM和IFN-γ含量显著高于SP组和LP组(P<0.05),血清IgG含量显著高于SP组(P<0.05)。
表2 饲粮添加糖萜素和植物乳杆菌对肉鹅血清免疫指标的影响

Table 2 Effects of dietary supplementation of saccharicter-penin and Lactobacillus plantarum on serum immune indices of meat geese

项目Items 干扰素-γ IFN-γ/(μg/mL) 免疫球蛋白M IgM/(μg/mL) 免疫球蛋白G IgG/(g/L)
组别Groups
CON 206.74b 718.29c 857.60b
SP 214.18b 815.56b 905.89b
LP 200.58b 768.58bc 1 013.60a
SP+LP 236.81a 975.91a 1 046.09a
SEM 3.65 22.07 21.76
P值P-value <0.01 <0.01 <0.01

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)。表3和表5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 Table 3 and Table 5.

2.2 饲粮添加糖萜素和植物乳杆菌对肉鹅空肠细胞因子含量的影响

由表4可知,与CON组相比,SP组、LP组和SP+LP组的空肠IL-1β、IL-2和TNF-α含量显著降低(P<0.05),空肠IL-4和sIgA含量显著提高(P<0.05)。此外,SP+LP组的空肠IL-4含量显著高于SP组和LP组(P<0.05),空肠TNF-α含量显著低于SP组和LP组(P<0.05)。
表3 饲粮添加糖萜素和植物乳杆菌对肉鹅空肠细胞因子含量的影响

Table 3 Effects of dietary supplementation of saccharicter-penin and Lactobacillus plantarum on jejunum cytokine contents of meat geese

项目
Items		 白细胞介素-1β
IL-1β/(μg/mL)		 白细胞介素-2
IL-2/(μg/mL)		 白细胞介素-4
IL-4/(ng/L)		 分泌型免疫球蛋白A
sIgA/(μg/mL)		 肿瘤坏死因子-α
TNF-α/(ng/L)
组别Groups
CON 42.02a 19.06a 173.15d 7.57c 308.01a
SP 36.91b 17.33b 181.62c 8.44b 290.32b
LP 35.48b 18.26c 199.38b 8.79a 279.17b
SP+LP 35.23b 17.12b 222.47a 9.11a 232.92c
SEM 0.69 0.19 4.08 0.13 6.12
P值P-value 0.01 <0.01 <0.01 <0.01 <0.01
表4 饲粮添加糖萜素和植物乳杆菌对肉鹅肠道形态的影响

Table 4 Effects of dietary supplementation of saccharicter-penin and Lactobacillus plantarum on intestinal morphology of meat geese

项目
Items		 组别Groups SEM P值
P-value
CON SP LP SP+LP
空肠
Jejunum		 绒毛高度Villus height/μm 366.91c 417.20b 395.72b 533.02a 12.12 <0.01
隐窝深度Crypt depth/μm 155.33a 152.98a 128.39b 108.57c 4.32 <0.01
绒毛高度/隐窝深度
Villus height/crypt depth		 2.38c 2.76bc 3.10b 4.99a 0.20 <0.01
回肠
Ileum		 绒毛高度Villus height/μm 400.90b 417.69b 480.63a 493.43a 8.29 <0.01
隐窝深度Crypt depth/μm 153.85a 119.01b 110.61b 102.05b 4.24 <0.01
绒毛高度/隐窝深度
Villus height/crypt depth		 2.64c 3.56b 4.42a 4.74a 0.10 <0.01

同行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)。表6和表7同。

In the same row, 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 Table 6 and Table 7.

2.3 饲粮添加糖萜素和植物乳杆菌对肉鹅肠道形态的影响

由表4可知,与CON组相比,SP组、LP组和SP+LP组的空肠绒毛高度显著提高(P<0.05);LP组和SP+LP组的空肠隐窝深度显著降低(P<0.05),空肠绒毛高度/隐窝深度显著提高(P<0.05)。此外,SP+LP组的空肠绒毛高度和绒毛高度/隐窝深度显著高于SP组和LP组(P<0.05),空肠隐窝深度显著低于SP组和LP组(P<0.05)。
与CON组相比,LP组和SP+LP组的回肠绒毛高度显著提高(P<0.05);SP组、LP组和SP+LP组的回肠隐窝深度显著降低(P<0.05),回肠绒毛高度/隐窝深度显著提高(P<0.05)。此外,SP+LP组的回肠绒毛高度和绒毛高度/隐窝深度显著高于SP组(P<0.05)。

2.4 饲粮添加糖萜素和植物乳杆菌对肉鹅肠道菌群结构的影响

2.4.1 样本测序数据质量评估

操作分类单元(OTU)绘制的维恩图如图1-A所示。Con组、SP组、LP组和SP+LP组分别包含176、116、107和76个独特序列,4组共有1 358个序列。物种多样性曲线和物种积累箱线图如图1-B、图1-C所示,用于计算物种丰富度和多样性。当测序数量达到20个时,物种积累箱线图趋于平坦,表明OTU序列足以预测样本的物种丰富度。同时,物种多样性曲线也趋于渐近,证明序列的深度足以代表大部分物种丰富度和细菌群落多样性。
图1 饲粮添加糖萜素和植物乳杆菌对肉鹅盲肠菌群多样性和丰富度的影响

A:操作分类单元绘制的维恩图 Venn diagram drew by OTU;B:物种多样性曲线 species diversity curve;C:物种累积箱线图 species accumulation box diagram。

CON:对照组 CON group;SP:糖萜素组 SP group;LP:植物乳杆菌组 LP group;SP+LP:糖萜素+植物乳杆菌组 SP+LP group。

Fig.1 Effects of dietary supplementation of saccharicter-penin and Lactobacillus plantarum on cecal flora diversity and richness of meat geese

2.4.2 Alpha多样性指数

由表5可知,使用Chao1、ACE、Shannon和Simpson指数来估计粪便微生物组分类群的丰度和多样性,与CON组相比,SP组、LP组、SP+LP组的Chao1指数提高(P>0.05),说明微生物丰富度增加;SP组和SP+LP组的Shannon和Simpson指数降低(P>0.05),说明微生物多样性增加。
表5 饲粮添加糖萜素和植物乳杆菌对肉鹅盲肠菌群α多样性指数的影响

Table 5 Effects of dietary supplementation with saccharicter-penin and Lactobacillus plantarum on cecal flora α diversity indexes of meat geese

项目
Items		 ACE指数
ACE index		 Chao1指数
Chao1 index		 Shannon指数
Shannon index		 Simpson指数
Simpson index
组别Groups
CON 1 516.87 1 518.24 6.92 0.976
SP 1 614.56 1 594.14 6.66 0.956
LP 1 580.46 1 559.67 6.80 0.970
SP+LP 1 742.42 1 728.30 7.00 0.968
SEM 60.66 60.70 0.12 0.004
P值P-value 0.676 0.642 0.789 0.425

2.4.3 物种相对丰度柱状图

如图2所示,在门水平上,排名前10位的物种为拟杆菌门、厚壁菌门、变形菌门、弯曲菌门、疣微菌门、放线菌门、蓝菌门、铁细菌门、脱铁杆菌门和梭杆菌门,其中厚壁菌门和拟杆菌门为优势门。由表6可知,与对照组相比,LP组和SP+LP组的盲肠拟杆菌门相对丰度显著增加(P<0.05),SP组、LP组和SP+LP组的盲肠弯曲菌门相对丰度显著降低(P<0.05)。
图2 饲粮添加糖萜素和植物乳杆菌对肉鹅盲肠物种相对丰度的影响(门水平)

Fig.2 Effects of dietary supplementation of saccharicter-penin and Lactobacillus plantarum on relative abundance of cecal species of meat geese (phylum level)

表6 饲粮添加糖萜素和植物乳杆菌对肉鹅盲肠微生物在门水平的优势菌群的影响

Table 6 Effects of dietary supplementation of saccharicter-penin and Lactobacillus plantarum on dominant microflora in cecum of meat geese at phylum level %

项目
Items		 组别Groups SEM P值
P-value
CON SP LP SP+LP
拟杆菌门Bacteroidota 50.49b 56.22ab 64.04a 60.99a 1.84 0.04
弯曲菌门Campilobacterota 2.33a 0.39b 0.043b 0.022b 0.01 <0.01
如图3所示,在属水平上,排名前10的物种为拟杆菌属、未分类菌属、另枝菌属、脱硫弧菌属、巴恩斯氏菌属、普雷沃氏菌属、未分类拟杆菌科、普氏栖粪杆菌属、巨单细胞菌属、罕见小球菌属。由表7可知,与对照组相比,LP组的盲肠拟杆菌属相对丰度显著增加(P<0.05),LP组和SP+LP组的盲肠巴恩斯氏菌属相对丰度显著升高(P<0.05),SP+LP组的盲肠普氏栖粪杆菌菌属相对丰度显著升高(P<0.05)。
图3 饲粮添加糖萜素和植物乳杆菌对肉鹅盲肠物种相对丰度的影响(属水平)

Fig.3 Effects of dietary supplementation with saccharicter-penin and Lactobacillus plantarum on the relative abundance of cecal species of meat geese (genus level)

表7 饲粮添加糖萜素和植物乳杆菌对肉鹅盲肠微生物在属水平的优势菌群的影响

Table 7 Effects of dietary supplementation of saccharicter-penin and Lactobacillus plantarum on dominant microflora in cecum of meat geese at genus level %

项目
Items		 组别Groups SEM P值
P-value
CON SP LP SP+LP
拟杆菌属Bacteroides 29.91b 30.15b 37.57a 35.11ab 1.24 0.04
巴恩斯氏菌属Barnesiella 2.17c 3.06bc 9.82a 5.82b 0.84 <0.01
普氏栖粪杆菌属Faecalibacterium 1.62b 2.40b 1.75b 5.11a 0.52 0.04

3 讨论

3.1 饲粮添加糖萜素和植物乳杆菌对肉鹅免疫功能的影响

血清免疫球蛋白含量是反映动物体液免疫状态的重要参数,与动物机体免疫功能和抵抗各类感染有关[22]。研究表明,皂苷可以提高免疫球蛋白的输出,并进一步诱导细胞和体液免疫[23]。Han等[24]研究发现,饲粮中添加植物乳杆菌可以显著提高肉鸡血液IgG含量。Song等[7]研究发现,饲粮中添加植物乳杆菌可以显著提高肉鸡血中IgM和IFN-γ含量。在本试验中,饲粮中糖萜素和植物乳杆菌联合使用时,血清IgM、IFN-γ含量高于单独添加糖萜素或植物乳杆菌,且免疫效果优于单独添加,其结果可能是由于糖萜素和植物乳杆菌的作用机制,刺激肠道免疫细胞的活性,增强机体免疫反应[25-26];其糖萜素和植物乳杆菌产生显著的交互作用,可能是由于糖萜素对肉鹅肠道中植物乳杆菌的生长有促进作用[27],进而影响机体免疫功能。

3.2 饲粮添加糖萜素和植物乳杆菌对肉鹅肠道免疫因子含量的影响

细胞因子在肠道炎症反应中起重要作用,机体免疫状态可以通过免疫细胞因子水平直接反映出来。Wang等[28]研究发现,饲粮中添加植物乳杆菌B1显著增加了肉鸡回肠黏膜sIgA含量,并显著降低肉鸡回肠IL-2、IL-4和TNF-α含量。Cao等[29]研究发现,饲粮中添加植物乳杆菌降低了肉鸡的促炎细胞因子含量,并改善了肉鸡的抗氧化能力。Dai等[30]研究发现,饲粮中添加皂苷可以显著降低肉鸡血清TNF-α含量。在本试验中,饲粮中糖萜素和植物乳杆菌联合使用时,IL-1β、IL-2和TNF-α含量低于单独添加糖萜素或植物乳杆菌;IL-4和sIgA含量高于单独添加糖萜素或植物乳杆菌,表明两者联合使用对提高肉鹅免疫功能可能更具优势,这种作用可能是通过植物乳杆菌和糖萜素协同刺激体液和细胞免疫引起的[23,31]。与以往研究不同的原因可能是动物种类、品种、益生菌种类或检测点不同。

3.3 饲粮添加糖萜素和植物乳杆菌对肉鹅肠道形态的影响

绒毛高度、隐窝深度、绒毛高度/隐窝深度是肠道形态学的重要参数,是衡量肠道健康和功能的重要指标[32]。糖萜素具有改善肠道对营养物质吸收的功能,并可改善动物肠道屏障[33]。Youssef等[34]研究表明,饲粮中添加皂苷可以使肉鸡小肠形态得到改善。皂苷作为糖萜素中主要成分,可以减少有害菌,改善肠道菌群,进而改善肠道形态。Han等[24]研究表明,饲粮中添加植物乳杆菌增加了肉鸡空肠绒毛高度并降低了隐窝深度。绒毛高度的增加和绒毛高度与隐窝深度的比率可能与有益细菌(乳酸杆菌)数量的增加有关[35],其具体机制还需要进一步研究。本试验研究表明,饲粮中糖萜素和植物乳杆菌联合使用时,在空肠绒毛高度、绒毛高度/隐窝深度及回肠隐窝深度、绒毛高度/隐窝深度比单一添加糖萜素或植物乳杆菌具有更多优势。

3.4 饲粮添加糖萜素和植物乳杆菌对肉鹅肠道菌群结构的影响

肠道微生物菌群的平衡与稳定在畜禽生长发育、营养消化和免疫拮抗等方面起着有着重要作用[36]。抗菌肽(HDP)具有抗菌和免疫特性[37],丁酸盐、维生素D3等可以诱导HDP分泌[38]。肠道微生物产生的丁酸盐是唯一能够诱导HDP表达的微生物刺激物[39],其中肠道中普氏栖粪杆菌已经被证明会产生丁酸盐[40]。有研究表明植物乳杆菌可以诱导HDP表达,防止病菌侵染[41]。在本试验中,饲粮中糖萜素和植物乳杆菌联合使用时,上调了盲肠巴恩斯氏菌属和普氏栖粪杆菌属相对丰度,下调了变形菌门相对丰度。因此,糖萜素和植物乳杆菌联用,可能会提高丁酸盐含量进而影响HDP表达。

4 结论

① 饲粮中糖萜素和植物乳杆菌联合使用时,对血清IgM、IFN-γ含量呈协同效应,且提高肉鹅免疫效果优于单独添加。
② 饲粮中单一添加糖萜素和植物乳杆菌均可显著降低肉鹅空肠IL-1β、IL-2和TNF-α含量,显著增加肉鹅空肠IL-4和sIgA含量,二者联合使用具有协同作用。
③ 饲粮中单一添加糖萜素和植物乳杆菌能够促进肠道发育,二者联合使用具有协同作用。
④ 饲粮中单一添加糖萜素和植物乳杆菌,在门水平上,盲肠拟杆菌门相对丰度显著增加,盲肠弯曲菌门相对丰度显著降低;在属水平上,盲肠拟杆菌属、巴恩斯氏菌属和普氏栖粪杆菌属相对丰度升高,优化了肠道菌群结构。并且,二者联合使用具有协同作用。

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