畜牧兽医学报 ›› 2023, Vol. 54 ›› Issue (11): 4676-4690.doi: 10.11843/j.issn.0366-6964.2023.11.022

• 营养与饲料 • 上一篇    下一篇

枯草芽孢杆菌对脂多糖应激肉仔鸡肠道免疫、肠道组织形态以及肠道屏障的影响

秦士贞, 杨敏敏, 任志雄, 李金录, 唐德富, 史兆国*   

  1. 甘肃农业大学动物科学技术学院, 兰州 730070
  • 出版日期:2023-11-23 发布日期:2023-11-26
  • 通讯作者: 史兆国,主要从事家禽生产与营养研究,E-mail:1991378985@qq.com
  • 作者简介:秦士贞(1988-),女,甘肃平川人,副教授,硕士生导师,主要从事家禽营养研究,E-mail:qinshizhen@163.com
  • 基金资助:
    甘肃农业大学学科团队项目(GAU-XKTD-2022-24);国家自然科学基金(31960668)

Effects of Bacillus subtilis on Intestinal Immunity, Intestinal Tissue Morphology and Intestinal Barrier of Broilers Challenged with Lipopolysaccharide

QIN Shizhen, YANG Minmin, REN Zhixiong, LI Jinlu, TANG Defu, SHI Zhaoguo*   

  1. College of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070, China
  • Online:2023-11-23 Published:2023-11-26

摘要: 本试验旨在研究枯草芽孢杆菌(BS)对脂多糖(LPS)应激肉仔鸡肠道免疫、肠道组织形态和肠道屏障的影响。选用1日龄健康爱拔益加(AA)肉仔鸡240只(公母各占1/2),采用2×2两因子完全随机试验设计,两因子分别为:日粮处理(基础日粮或基础日粮+200 g·t-1 BS的试验日粮)和应激处理(口腔灌服生理盐水或LPS)。即试验分为:1)基础日粮+生理盐水组;2)基础日粮+LPS组;3)试验日粮+生理盐水;4)试验日粮+LPS组,共计4个组,每组6个重复,每个重复10只鸡,试验期为28 d,分为3个阶段,包括应激前期(1~14 d)、 LPS 应激期(15~21 d)和恢复期(22~28 d)。结果表明:1)在第21天,BS日粮与LPS应激对十二指肠白细胞介素-1β(Interleukin-1β, IL-1β)、Toll样受体4 (Toll like rececptor 4, TLR4)mRNA的表达量有显著的交互作用(P<0.05),对空肠髓样分化因子88(Myeloid differentiation factor 88, MyD88)mRNA的表达量有显著性交互作用(P<0.05),对回肠肿瘤坏死因子-α(Tumor necrosis actor-α, TNF-α)mRNA的表达量有显著性交互作用(P<0.05)。在无LPS应激条件下,BS日粮对上述指标均无显著影响(P> 0.05);在LPS应激条件下,BS显著下调以上基因的 mRNA的表达量(P<0.05)。2)BS日粮与LPS应激对21 d十二指肠VH及V/C的比值和空肠CD有显著性交互作用(P<0.05)。不管有无LPS应激,BS日粮均可显著提高十二指肠VH及V/C的比值(P<0.05);在无LPS应激条件下,BS日粮显著降低了空肠CD(P<0.05);在LPS应激条件下,BS日粮对各肠段CD无显著影响(P>0.05)。在第28天,BS日粮与LPS应激对空肠VH和V/C的比值有显著性交互作用(P<0.05)。无LPS应激条件下,BS日粮对空肠VH和V/C的比值无显著影响(P>0.05);LPS应激条件下,BS日粮显著增加了空肠VH和V/C的比值(P<0.05)。3)LPS应激显著降低了21 d各肠段Claudin-1、Occludin、ZO-1和MUC2 mRNA的表达量(P<0.05)。BS日粮显著提高了十二指肠和Occludin、ZO-1和空肠Occludin、ZO-1MUC2 mRNA的表达量(P<0.05)。结果显示,日粮中添加BS可显著降低小肠促炎细胞因子的表达,提高紧密连接蛋白Claudin-1、Occludin、ZO-1和MUC2 mRNA的表达量,增加肠道VH及V/C的比值,有效缓解了免疫应激对肠道形态结构的损伤,促进了小肠的生长发育,维持了肠道的健康。

关键词: 枯草芽孢杆菌, 肉仔鸡, 脂多糖, 肠道屏障

Abstract: The purpose of this experiment was to investigate the effects of Bacillus subtilis (BS) on intestinal immunity, intestinal tissue morphology, and intestinal barrier of broilers challenged with lipopolysaccharide (LPS). A total of 240 one-day-old healthy AA broilers (males and females in half) were randomly divided to 4 treatments as a 2×2 factorial design. The main factors were diets (a basic diet or a experimental diet added with 200 g·t-1 BS) and LPS stress (oral administration of LPS solution or saline),and randomly divided into: 1) The basal diet+normal saline group; 2) The basal diet+LPS group; 3) The experimental diet+ normalsaline group; 4) The experimental diet+LPS group, respectively, a total of 4 groups, each group composed of 6 replicates with 10 broilers per replicate. The trial lasted for 28 days, and the experimental period was divided into three stages, including pre-stress (1 to 14 days), LPS stress (15 to 21 days) and recovery (22 to 28 days). The results showed as follows: l ) On the 21st day, there was a significant interaction between BS diet and LPS stress on the expression of IL-1β and TLR4 mRNA in duodenum(P<0.05), MyD88 mRNA in jejunum(P<0.05), and TNF-α mRNA in ileum(P<0.05). Without LPS stress, none of the above indicators were significantly affected by BS diet (P>0.05). Under LPS stress, BS significantly down-regulated the mRNA expression levels of those genes(P<0.05).2)On the 21st day, there was an significant interaction between BS and LPS on VH and V/C ratio in duodenum and CD in jejunum (P<0.05). BS diet significantly increased VH and V/C ratios with or without LPS stress in duodenum. Without LPS stress, BS diet significantly reduced CD of jejunum (P<0.05); Under LPS stress, BS diet had no significant effect on CD of each intestinal segment(P>0.05). On the 28th day, there existed significant interaction between BS and LPS on VH and V/C ratio in jejunum. Without LPS stress, BS diet has no significant effect on VH or V/C ratio of jejunum (P>0.05); Under LPS stress, the BS diet significantly increased jejumum VH and V/C ratio (P<0.05).3)LPS stress significantly reduced the mRNA expression of Claudin-1, Occludin, ZO-1 and MUC2 in the intestine at 21 days. BS diet significantly increased the expression of Occludin and ZO-1 mRNA in duodenum, as well as Occludin, ZO-1 and MUC2 mRNA in jejunum. The results showed that dietary BS supplementation significantly decreased the expression of intestinal proinflammatory cytokines, increased the expression of tight junction proteins Claudin-1, Occludin, ZO-1 and MUC2 mRNA, increased intestinal VH and V/C ratio, effectively alleviated the damage of immune stress on intestinal morphological structure, and promoted the growth and development of small intestine,maintains intestinal health. Dietary BS can alleviate the damage of intestinal morphology and structure caused by LPS stress, promote the growth and development of small intestine, and maintain intestinal barrier.

Key words: Bacillus subtilis, broilers, lipopolysaccharide, intestinal barrier

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