仔猪空肠和回肠微生物多样性分析

Analysis of intestinal microbial diversity in piglet jejunum and ileum

[目的]微生物作为影响机体营养物质消化吸收的重要因素,在仔猪肠道生长发育过程中起至关重要的作用。本研究分析了马身猪与晋汾白猪仔猪在生长发育时期的空肠、回肠微生物组成变化,旨在揭示肠道菌群与血清生化指标和肠道结构的关系。[方法]收集马身猪和晋汾白猪在3个发育时期(1日龄,28日龄和70日龄)仔猪空肠、回肠组织以及内容物(n=3),通过16S rRNA高通量测序与生信技术分析微生物多样性,HE染色测定肠道形态结构,相关性分析将微生物菌群与血清免疫指标和肠绒毛结构关联。[结果]仔猪初生期小肠绒毛最长,保育末期隐窝深度最高。晋汾白猪仔猪肠道微生物分布于28个门和518个属;马身猪仔猪肠道微生物分布于19个门和325个属。其中,厚壁菌门、拟杆菌门和变形菌门为仔猪优势菌门,且乳酸杆菌丰度均较高。此外,仔猪肠道微生物多样性与血清免疫指标(LPS、TNF⁃α、IL⁃6、DAO、D⁃LA)以及隐窝深度显著相关,而与绒毛长度显著不相关。[结论]地方品种马身猪仔猪肠道微生物组成更加保守,乳酸杆菌、Ferruginibacter等在猪抗逆性能方面发挥重要作用,Quinella、Haliangium等在营养吸收代谢具有重要作用。本研究可为地方品种与培育品种仔猪小肠微生物多样性比较以及微生物对宿主的影响研究奠定基础。

[Objective]Microorganisms can affect the body′s digestion and absorption of nutrients,therefore the intestinal microorganisms play the important role in the development and growth of piglets by affecting the digestion and absorption of nutrients in the body.The aims of this study were to analyze the variation of jejunum and ileum microflora composition of Mashen and Jinfen White piglets at three different developmental stages,and to explore the potential relationship between intestinal microbiota and intestinal development.[Methods]In this study,Mashen and Jinfen White piglets were used as experimental animals.Three boars with similar body weight were randomly selected at three developmental points including at the birth(1 d),the end of lactation(28 d)and the end of nursing(70 d).The microbial diversity of jejunum and ileum contents was analyzed using 16S rRNA sequencing technology and bioinformatic analysis.The intestinal morphology and structure were determined by hematoxylin-eosin staining(HE),and the correlation of intestinal microflora with serum immune indexes and intestinal villus structure was analyzed using redundancy analysis(RDA)methods.[Results]Results indicated that the piglets had the longest intestinal villus at the initial stage and the highest crypt depth at the end of the nursery stage in both breeds.The intestinal microbes of Jinfen White piglets were distributed in 28 phyla and 518 genera and that of Mashen piglets were distributed in 19 phyla and 325 genera by bacteria taxonomy analysis.Among them,Firmicutes,Proteobacteria and Bacteroides were the dominant phyla,and the abundance of Lactobacillus was high in the intestine of both breeds.Correlation analysis showed that intestinal microbial diversity was significantly correlated with serum immune indexes including LPS,TNF-α,IL-6,DAO,D-LA,and crypt depth(P<0.05),but not with the villus length(P>0.05).[Conclusion]In summary,it was shown that the gut microbial composition of native breeds(Mashen)piglet was more conservative.Lactobacillus,Ferruginibacter,Thiopseudomonas,Thauera,Mucispirillum play important roles in resistance performance of piglets,and Quinella,Haliangium,Zoogloea,Anoxybacillus,and Lachnospiraceae were of importance in nutrient absorption and metabolism.The results of this study can provide reference for the research on intestinal microbiota to host and healthy feeding in piglets.