动物营养学报 >

2022 , Vol. 34 >Issue 8: 4944 - 4958

DOI: https://doi.org/10.3969/j.issn.1006-267x.2022.08.020

研究论文 RESEARCH PAPER

饲粮添加迷迭香酸对产肠毒素大肠杆菌K88攻毒断奶仔猪结肠菌群组成、屏障功能及炎症反应的影响

  • 雷铭康 ,
  • 李润林 ,
  • 李盼盼 ,
  • 吴佳庆 ,
  • 汪晶 ,
  • 朱伟云
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  • 1. 南京农业大学动物科技学院, 南京 210095;
    2. 国家动物消化道营养国际联合研究中心, 南京 210095
雷铭康(1997—),男,山东潍坊人,硕士研究生,从事消化道微生物研究。E-mail:2019105062@njau.edu.cn

收稿日期: 2022-01-12

  网络出版日期: 2022-08-11

基金资助

国家重点研发计划资助(2017YEF0135200)

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Effects of Dietary Rosmarinic Acid on Colonic Microbiota Composition, Barrier Function and Inflammatory Response in Weaned Piglets Challenged with Enterotoxigenic Escherichia coli K88

  • LEI Mingkang ,
  • LI Runlin ,
  • LI Panpan ,
  • WU Jiaqing ,
  • WANG Jing ,
  • ZHU Weiyun
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  • 1. College of Animal Science and Technology, Nanjing Agricultural University, Nanjing 210095, China;
    2. National Center for International Research on Animal Gut Nutrition, Nanjing 210095, China

Received date: 2022-01-12

  Online published: 2022-08-11

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摘要

本研究旨在探究饲粮添加迷迭香酸(RA)对产肠毒素大肠杆菌K88(ETEC K88)攻毒断奶仔猪结肠菌群组成、屏障功能及炎症反应的影响。选取18头体重为(6.91±1.02) kg的健康21日龄断奶仔猪,随机分为对照组(CON组)、攻毒组(K88组)和迷迭香酸组(RA组),每组6头仔猪。CON组和K88组每天饲喂基础饲粮,RA组每天饲喂含RA的试验饲粮(在基础饲粮基础上添加500 mg/kg RA)。试验第19~20天,K88组、RA组仔猪每天灌喂4 mL浓度为109 CFU/mL的ETEC K88重悬液攻毒,对照组仔猪则每天灌喂同等剂量的无菌生理盐水。所有仔猪在试验第22天屠宰取样。结果显示:1)与CON组相比,K88组断奶仔猪腹泻指数显著增加(P<0.05);与K88组相比,RA组断奶仔猪腹泻率极显著降低(P<0.01)。2)与K88组相比,RA组断奶仔猪结肠隐窝深度和黏膜厚度极显著降低(P<0.01)。3)与CON组相比,K88组断奶仔猪结肠菌群Chao1和ACE指数显著降低(P<0.05)。4)在门水平上,与K88组相比,RA组断奶仔猪结肠菌群中拟杆菌门(Bacteroidetes)的相对丰度极显著降低(P<0.01),变形菌门(Proteobacteria)的相对丰度显著降低(P<0.05),放线菌门(Actinobacteria)的相对丰度显著增加(P<0.05);在属水平上,与K88组相比,RA组断奶仔猪结肠菌群中克里斯滕森菌科R-7群(Christensenellaceae_R-7_group)的相对丰度极显著增加(P<0.01),布劳特氏菌属(Blautia)、粪球菌属3(Coprococcus_3)、双歧杆菌属(Bifidobacterium)的相对丰度显著增加(P<0.05),而脱硫弧菌属(Desulffovibrio)、埃希氏菌-志贺氏菌属(Escherichia-Shigella)的相对丰度显著降低(P<0.05)。5)与K88组相比,RA组断奶仔猪结肠食糜中乙酸、丁酸和总短链脂肪酸含量均显著增加(P<0.05)。6)与K88组相比,RA组断奶仔猪结肠黏膜中闭锁蛋白(Occludin)、闭锁小带蛋白-1(ZO-1)的mRNA相对表达量极显著增加(P<0.01)。7)与K88组相比,RA组断奶仔猪结肠黏膜中白细胞介素(IL)-1β、肿瘤坏死因子-α(TNF-α)的含量极显著降低(P<0.01),IL-10含量极显著增加(P<0.01),并且结肠黏膜中Toll样受体(TLR)-2(P<0.01)、TLR-4(P<0.01)、TLR-5(P<0.05)、核因子-κB (NF-κB)(P<0.01)的mRNA相对表达量显著或极显著降低。综上可知,饲粮添加RA可以改善ETEC K88攻毒断奶仔猪的结肠形态结构,调节结肠菌群组成,提高结肠食糜中细菌代谢产物短链脂肪酸的含量,有利于维持结肠屏障功能,缓解ETEC K88攻毒引起的结肠炎症反应,缓解断奶仔猪腹泻,维持肠道健康。

关键词： 迷迭香酸; 产肠毒素大肠杆菌K88; 断奶仔猪; 结肠; 菌群; 屏障功能; 炎症反应

本文引用格式

雷铭康 , 李润林 , 李盼盼 , 吴佳庆 , 汪晶 , 朱伟云 . 饲粮添加迷迭香酸对产肠毒素大肠杆菌K88攻毒断奶仔猪结肠菌群组成、屏障功能及炎症反应的影响[J]. 动物营养学报, 2022 , 34(8) : 4944 -4958 . DOI: 10.3969/j.issn.1006-267x.2022.08.020

Abstract

This study aimed to investigate the effects of dietary rosmarinic acid (RA) on the colonic microbiota composition, barrier function and inflammatory response in weaned piglets challenged with enterotoxigenic Escherichia coli K88 (ETEC K88). Eighteen healthy piglets with the body weight of (6.91±1.02) kg, weaned at the age of 21 days, were randomly divided into the control group (CON group), challenged group (K88 group) and RA group, with 6 piglets in each group. Piglets in the CON group and K88 group were fed a basal diet every day, and piglets in the RA group were fed an experimental diet (adding 500 mg/kg RA in the basal diet) every day. On the 19th to 20th day of the experiment, piglets in the K88 group and RA group were challenged by oral administration of 4 mL 109 CFU/mL ETEC K88 suspension every day, while piglets in the CON group were given an equal volume of normal saline. All piglets were slaughtered and sampled on the 22nd day of the experiment. The results showed as follows: 1) the diarrhea index of weaned piglets in K88 group was significantly increased compared with CON group (P<0.05). The diarrhea rate of weaned piglets in RA group was extremely significantly decreased compared with K88 group (P<0.01). 2) The colonic crypt depth and mucosal thickness were extremely significantly decreased in RA group compared with K88 group (P<0.01). 3) The Chao1 and Ace indexes of microbiota in colon of weaned piglets were significantly decreased in K88 group compared with CON group (P<0.05). 4) At the phylum level, the relative abundance of Bacteroides was extremely significantly decreased (P<0.01), and the relative abundance of Proteobacteria was significantly decreased (P<0.05), but the relative abundance of Actinobacteria was significantly increased (P<0.05) in colonic microbiota of weaned piglets in RA group piglets compared with K88 group. At the genus level, compared with K88 group, the relative abundance of Christensenellaceae_R-7_group was extremely significantly increased (P<0.01), the relative abundances of Blautia, Coprococcus_3, and Bifidobacterium were significantly increased (P<0.05), but the relative abundances of Desulfovibrio and Escherichia-Shigella were significantly decreased in colonic microbiota of weaned piglets in RA group (P<0.05). 5) The contents of acetic acid, butyric acid and total short chain fatty acids in colonic chyme were significantly increased in RA group compared with K88 group (P<0.05). 6) The mRNA relative expression levels of Occludin and zonula occluden-1 (ZO-1) were extremely significantly increased in colonic mucosa of weaned piglets in RA group compared with K88 group (P<0.01). 7) The contents of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) in the colonic mucosa of weaned piglets were extremely significantly decreased (P<0.01), while the content of interleukin-10 (IL-10) extremely was significantly increased in RA group piglets compared with K88 group (P<0.01). And the mRNA relative expression levels of Toll-like receptor (TLR)-2 (P<0.01), TLR-4 (P<0.01), TLR-5 (P<0.05), nuclear factor-κB (NF-κB) (P<0.01) in colonic mucosa of weaned piglets were significantly or extremely significantly decreased in RA group compared with K88 group. In conclusion, dietary RA improves the colonic morphology, modulates the colonic microbiota composition, increases the contents of bacterial metabolites——short chain fatty acids in colonic chyme, maintains the intestinal barrier function, alleviates the colonic inflammatory response and diarrhea of weaned piglets challenged by ETEC K88, and then keeps the intestinal health.

Key words： rosmarinic acid; ETEC K88; weaned piglets; colon; microbiota; barrier function; inflammatory response

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