基于MiSeq高通量测序方法研究酵母壁多糖对断奶仔猪盲肠菌群结构的影响

Effect of Yeast Cell Wall Polysaccharides on Cecum Bacteria Structure of Weaned Piglets by MiSeq Sequencing Technology

本试验旨在探讨饲粮中添加酵母壁多糖对断奶仔猪盲肠菌群结构的影响。采用单因素试验设计,选取180头遗传背景相同、胎次相近、体重接近的21日龄断奶仔猪,随机分为4个处理,每个处理5个重复,每个重复9头猪。4个处理分别饲喂0%酵母壁多糖饲粮(对照组)、0.15%酵母壁多糖饲粮(0.15%组)、0.30%酵母壁多糖饲粮(0.30%组)和0.45%酵母壁多糖饲粮(0.45%组)。试验期为21 d。取盲肠内容物提取细菌DNA,PCR扩增获得16S r RNA基因V4标签片段,进行Mi Seq高通量测序。结果表明:当酵母壁多糖添加量达一定程度时,仔猪盲肠菌群多样性会发生改变;在不考虑群落中每个物种丰度的前提下,0.30%酵母壁多糖组仔猪盲肠菌群最丰富;仔猪盲肠优势菌群为拟杆菌门、厚壁菌门、螺旋体门和变形菌门;与对照组相比,添加0.30%酵母壁多糖能降低仔猪肠道拟杆菌门含量(P<0.05),添加0.30%或0.45%酵母壁多糖能显著增加盲肠厚壁菌门含量(P<0.05),添加0.45%酵母壁多糖还能提高仔猪盲肠软壁菌门含量(P<0.05);酵母壁多糖具有降低盲肠变形菌门含量的趋势(P=0.066);普氏菌属为仔猪盲肠优势菌属,酵母壁多糖能显著提高仔猪盲肠厌氧弧菌属、瘤胃杆菌属、粪球菌属和考拉杆菌属的含量(P<0.05),显著降低链球菌属的含量(P<0.05)。由此可知,当酵母壁多糖添加量达到一定水平时,短期添加可提高断奶仔猪盲肠厚壁菌门以及瘤胃杆菌属和粪球菌属等优势菌群的含量。

An experiment was conducted to determine the effects of yeast cell wall polysaccharides on cecum bacteriastructure of weaned piglets. A total of one hundred and eighty piglets with the similar genetic background, health condition,parity and body weight which weaned at 21 days of age were randomly allotted to 4 groups with 5 replicated each and 9pigs in each replicate. Pigs in the four groups were fed control （without yeast cell wall polysaccharide）, 0.15% of yeastcell wall polysaccharide, 0.30% yeast cell wall polysaccharide and 0.45% yeast cell wall polysaccharide, respectively.The experiment lasted for 21 days. After 21 days, the piglets were killed and the samples of cecum content were collected.Total bacterial DNA of 20 samples of pigs per group was extracted, and used to amplify PCR for 16Sr RNA gene V4 tagfragment and sequence using high-throughput MiSeq technique. Based on the QIIME platform, the microbial communitydiversity in cecum content was compared. The results showed as follows： There were no significant differences onbacterial diversity in piglets receiving control diets and yeast cell wall polysaccharides. To a certain extent, dietarysupplementation yeast cell wall polysaccharides can change the bacterial diversity, and without considering the abundanceof each species, 0.30% yeast cell wall polysaccharide cecum flora was the most abundant. Bacteroidetes, Firmicutes,Spirochaetes and Proteobacteria were the dominant phyla in cecum content of piglets. Compared with the control group,dietary supplementation of 0.30% yeast cell wall polysaccharide significantly decreased Bacteroidetes （P〈0.05）, 0.30%and 0.45% yeast cell wall polysaccharides significantly increased the amounts of Firmicutes. Added 0.45% yeast cellwall polysaccharide can significantly increase the Tenericutes of cecum, in addition, Yeast cell wall polysaccharide couldreduce the Proteobacteria of cecum, but there were no significant differences （P=0.066）. Prevotella were the dominantgenus in cecum content of piglets. The yeast cell wall polysaccharides enhance（P〈0.05） Anaerovibrio, Ruminococcus,Coprococcus and Phascolarctobacterium, and reduced the Streptococcus of cecum（P〈0.05）. Accordingly, diets with yeastcell wall polysaccharides in short feeding-period could improve the cecum bacteria structure of weaned piglets. Diets withyeast cell wall polysaccharides could enhance the Advantage bacterium of cecum, such as Firmicutes, Ruminococcus,Coprococcus,etc.