羊源Lactobacillus plantarum对腹泻羔羊空肠菌群及肠道黏膜屏障的调控作用  

Sheep-derived Lactobacillus plantarum Regulates the Bacterial Community and Mucosal Barrier in Jejunum of Diarrheic Lambs

作　　者：李碧波 吴克 师晓龙 闫奕凝 李嘉豪 段国庆 李熊 任彦鹏 董佳宁 张春香[1] 任有蛇[1] LI Bibo;WU Ke;SHI Xiaolong;YAN Yining;LI Jiahao;DUAN Guoqing;LI Xiong;REN Yanpeng;DONG Jianing;ZHANG Chunxiang;REN Youshe(College of Animal Science,Shanxi Agricultural University,Taigu 030801,China)

机构地区：[1]山西农业大学动物科学学院,太谷030801

出　　处：《畜牧兽医学报》2024年第8期3552-3569,共18页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：山西省高等学校科技创新项目(2021L175);山西省博士毕业生、博士后研究人员来晋工作奖励资金科研项目(SXBYKY2021037);山西农业大学科技创新基金项目(2020BQ53);山西省现代农业产业技术体系建设(2023CYJSTX14)。

摘　　要：旨在研究Lactobacillus plantarum对腹泻羔羊血清抗氧化能力和空肠菌群及空肠黏膜屏障损伤的调控作用。本试验选用24只刚断奶的羔羊,采用完全随机试验设计随机分为4组,分别为对照组(C组)、腹泻组(D组)、腹泻+抗生素处理组(DA组)、腹泻+植物乳杆菌处理组(DL组),每组6个重复,在正式试验第1天开始对D组、DA组和DL组羔羊分别灌服20 mL ETEC K99菌液,持续8 d。在第9天早上,DA组羔羊注射氟苯尼考,DL组羔羊灌服植物乳杆菌菌液,持续4 d。试验第1、9、13天空腹采集羔羊血液用于检测羔羊血清抗氧化指标以及二胺氧化酶(DAO)和D-乳酸(D-LA)浓度;屠宰后采集羔羊空肠内容物以测定菌群组成,采集空肠组织用于检测肠道黏膜屏障相关蛋白和黏膜蛋白,阿利新蓝(AB)染色研究酸性黏液的变化。结果表明:1)大肠杆菌处理8 d后,D、DA和DL三组羔羊粪便均不成形,且三组羔羊血清中DAO和D-LA浓度显著高于C组(P<0.05),说明腹泻模型构建成功;2)在第9天,与C组相比,D、DA和DL组羔羊血清中T-AOC、SOD显著降低(P<0.05),MDA含量显著升高(P<0.05);与第9天相比,DA和DL组羔羊在第13天时其抗氧化能力显著增加(P<0.05),DAO、D-LA浓度和MDA显著降低(P<0.05);3)与C组相比,D组羔羊空肠组织Occludin、Claudin1和MUC2蛋白表达量显著下降(P<0.05);与D组相比,DA组羔羊空肠组织Occludin蛋白表达量显著提高(P<0.05),但Claudin1蛋白和MUC2蛋白的表达量有所降低(P>0.05);DL组羔羊空肠组织Occludin蛋白、Claudin1蛋白以及MUC2蛋白表达量显著高于DA组;4)与C组相比,D和DA组羔羊空肠内容物中菌群Chao值和Shannon值均显著降低(P<0.05),DL羔羊空肠内容物中菌群Chao值和Shannon值有所增加(P>0.05)。C、D和DL组羔羊空肠中Firmicutes丰度均显著高于DA组(P<0.05)。C、D和DA组Actinobacteriota丰度都显著高于DL组(P<0.05)。D组羔羊空肠中Proteobacteria丰度略高于C和DL组(P>0.05),DA组羔羊空肠中ProteobThe aim of the experiment was to investigate the mitigating regulating effect of Lactobacillus plantarum on diarrhea lambs in indexes including the damage of anti-oxidative capacity in serum,jejunal microbiota,and jejunal mucosal barrier damage in diarrhea lambs.[Methods]Twenty-four newly weaned lambs were selected and distributed into four groups using a completely randomized experimental design:the control group(C),the diarrhea group(D),the group of antibiotic-treated under diarrhea(DA),and the group of Lactobacillus plantarum-treated under diarrhea(DL),with six replicates in each group.Lambs in groups D,DA and DL were dosed with 20 mL of ETEC K99 starting on the first day of the formal trial for 8 d consecutively.On the morning of the ninth day,lambs in group DA were injected regulated with florfenicol,and lambs in group DL were regulated by dosing gavaged with Lactobacillus plantarum,and the regulation above treatment lasted for 4 consecutive days.The blood was collected fasting on days 1,9 and 13 of the trial for the detection of serum antioxidant indexes as well as diamine oxidase(DAO)and D-lactic acid(D-LA)concentration in lambs.After slaughter the jejunal content was collected to determine the composition of bacterial community.The jejunal mucosal were collected to explore the expression of intestinal mucosal barrier-associated proteins and mucus proteins.[Results]Alisin Blue staining was uesd to study the changes of acidic mucus.The results showed that:1)After 8 d of ETEC K99 treatment,the feces of lambs in D,DA and DL were all shapeless,and the concentrations of DAO and D-LA in the serum of lambs in the three groups were significantly higher than those in group C(P<0.05),which indicated that the model of diarrhea had been successfully constructed;2)On the 9 th day,compared with that of group C,the T-AOC and SOD in serum of lambs in D,DA and DL groups decreased significantly(P<0.05),and the MDA content increased significantly(P<0.05).Compared to on the 9 th day,the antioxidant capacity of lambs in DA and

关 键 词：植物乳杆菌 羔羊 抗氧化能力 肠道黏膜屏障 肠道菌群 

分 类 号：S858.26[农业科学—临床兽医学]