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母源喂养和配方奶喂养仔猪的肠道微生物基因表达。

Gut microbial gene expression in mother-fed and formula-fed piglets.

机构信息

Department of Surgery, The University of Chicago Pritzker School of Medicine, Chicago, Illinois, United States of America.

出版信息

PLoS One. 2010 Aug 27;5(8):e12459. doi: 10.1371/journal.pone.0012459.

Abstract

BACKGROUND

Effects of diet on the structure and function of gut microbial communities in newborn infants are poorly understood. High-resolution molecular studies are needed to definitively ascertain whether gut microbial communities are distinct in milk-fed and formula-fed infants.

METHODOLOGY/PRINCIPAL FINDINGS: Pyrosequencing-based whole transcriptome shotgun sequencing (RNA-seq) was used to evaluate community wide gut microbial gene expression in 21 day old neonatal piglets fed either with sow's milk (mother fed, MF; n = 4) or with artificial formula (formula fed, FF; n = 4). Microbial DNA and RNA were harvested from cecal contents for each animal. cDNA libraries and 16S rDNA amplicons were sequenced on the Roche 454 GS-FLX Titanium system. Communities were similar at the level of phylum but were dissimilar at the level of genus; Prevotella was the dominant genus within MF samples and Bacteroides was most abundant within FF samples. Screened cDNA sequences were assigned functional annotations by the MG-RAST annotation pipeline and based upon best-BLASTX-hits to the NCBI COG database. Patterns of gene expression were very similar in MF and FF animals. All samples were enriched with transcripts encoding enzymes for carbohydrate and protein metabolism, as well as proteins involved in stress response, binding to host epithelium, and lipopolysaccharide metabolism. Carbohydrate utilization transcripts were generally similar in both groups. The abundance of enzymes involved in several pathways related to amino acid metabolism (e.g., arginine metabolism) and oxidative stress response differed in MF and FF animals.

CONCLUSIONS/SIGNIFICANCE: Abundant transcripts identified in this study likely contribute to a core microbial metatranscriptome in the distal intestine. Although microbial community gene expression was generally similar in the cecal contents of MF and FF neonatal piglets, several differentially abundant gene clusters were identified. Further investigations of gut microbial gene expression will contribute to a better understanding of normal and abnormal enteric microbiology in animals and humans.

摘要

背景

饮食对新生婴儿肠道微生物群落的结构和功能的影响知之甚少。需要高分辨率的分子研究来明确确定母乳喂养和配方奶喂养的婴儿的肠道微生物群落是否不同。

方法/主要发现:采用基于焦磷酸测序的全转录组鸟枪法测序(RNA-seq)评估了 21 天大的新生仔猪(母猪喂养,MF;n=4)或人工配方喂养(配方喂养,FF;n=4)的肠道微生物群落的全基因表达。从每个动物的盲肠内容物中提取微生物 DNA 和 RNA。将 cDNA 文库和 16S rDNA 扩增子在 Roche 454 GS-FLX Titanium 系统上测序。在门水平上,群落相似,但在属水平上不同;MF 样本中优势属为普雷沃氏菌属,FF 样本中最丰富的是拟杆菌属。筛选的 cDNA 序列通过 MG-RAST 注释管道和最佳-BLASTX-hits 到 NCBI COG 数据库进行功能注释。MF 和 FF 动物的基因表达模式非常相似。所有样本均富含碳水化合物和蛋白质代谢酶的转录物,以及参与应激反应、与宿主上皮结合和脂多糖代谢的蛋白质。两组的碳水化合物利用转录物通常相似。MF 和 FF 动物中氨基酸代谢(如精氨酸代谢)和氧化应激反应相关的几种途径中涉及的酶的丰度不同。

结论/意义:本研究中鉴定的丰富转录本可能有助于远端肠道微生物元转录组的核心。尽管 MF 和 FF 新生仔猪盲肠内容物中的微生物群落基因表达总体相似,但鉴定出了几个差异丰度的基因簇。对肠道微生物基因表达的进一步研究将有助于更好地了解动物和人类正常和异常的肠道微生物学。

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