金华猪和长白猪粪便微生物移植小鼠的肠道古菌结构分析

本试验旨在比较肥胖型的金华猪和瘦肉型的长白猪肠道古菌结构的差异,分析菌群移植小鼠肠道古菌结构。采集金华猪和长白猪的新鲜粪便,通过灌胃移植到经广谱抗生素处理过的小鼠肠道中,提取2种猪粪便及其移植小鼠的盲肠内容物总基因组DNA,使用古菌通用引物对古菌16S rRNA的V4区进行PCR扩增,通过Illumina Hiseq测序平台对扩增产物进行高通量测序,使用QIIME等软件对测序序列进行分析统计。结果表明:金华猪和长白猪粪便及其移植小鼠盲肠内容物中的古菌主要包含3个门,分别为奇古菌门(Thaumarchaeota)、广古菌门(Euryarchaeota)和泉古菌门(Crenarchaeota),其中金华猪粪便中广古菌门的丰度高于长白猪;在属水平上主要包含4个已知属,优势属为餐古菌属(Cenarchaeum)。二者移植小鼠的盲肠内容物古菌结构,在门水平上,移植小鼠的奇古菌门比例占70%以上;奇古菌门的古菌几乎全部来源于餐古菌属;在门和属的水平上,金华猪和长白猪粪便移植小鼠的盲肠古菌结构无显著差异(P0.05),但基于操作分类单元(OTU)水平的主成分分析显示出很好的组内聚集。由此可见,奇古菌门的餐古菌属是金华猪和长白猪粪便和其移植小鼠盲肠中的优势古菌;金华猪粪便中广古菌门的丰度高于长白猪粪便,且二者移植小鼠盲肠内容物之间古菌结构在OTU水平上存在一定的差异。

Intestinal Archaea Community Structure Analysis of Mice Transplanted with Jinhua and Landrace Pig Feces

This experiment was conducted to explore the differences of the intestinal archaea community structures between the obese breed Jinhua pig and the lean breed Landrace pig,and to investigate the changes of the archaea community structure during the inter-species fecal microbiota transplantation (FMT).The fresh feces of Jinhua and Landrace pigs were collected and introduced into broad spectrum antibiotics treated germ-free mice guts by gavage,respectively.The total genomic DNA was extracted from the pig fecal samples and the transplanted mice cecal contents,and was used as templates to amplify the V4 region of the 16S rRNA gene by PCR with the universal primers of archaea.The PCR products were then subjected to high-throughput sequencing on an Illumina HiSeq sequencing platform. The obtained sequences were subsequently processed and analyzed with a series of software such as QIIME.The results showed as follows: at the phylum level,three main phyla of archaea,Thaumarchaeota,Euryarchaeota,and Crenarchaeota,were found in pig fecal samples and transplanted mice cecal contents,with Thaumarchaeota as the dominant phylum;while at the genus level,five known genera of archaea were observed,with Cenarchaeum as the dominant genus.The abundance of phylum Euryarchaeota in Jinhua pig's feces was higher than that in Landrace pig's feces.Thaumarchaeota dominate the archaea community structure of transplanted mice cecal contents,with abundance of more than 70%,and Thaumarchaeota mainly consisted of Cenarchaeum.It was observed that no significant difference was exhibited between the two groups of mice at the phylum and genus levels (P>0.05),but the operational taxonomic unit (OTU) based principal coordinate analysis showed a good aggregation within each group.It is concluded that genus Cenarchaeum of phylum Thaumarchaeota is the dominant archaea in feces of Jinhua and Landrace pigs and cecal contents of mice recipients.Phylum Euryarchaeota is more abundant in Jinhua pig's feces than in Landrace pig's feces, and there is difference in archaea community structures between their respective mice recipients at the OTU level.