Changes in the diversity of pig ileal lactobacilli around weaning determined by means of 16S rRNA gene amplification and denaturing gradient gel electrophoresis
Changes in the diversity of pig ileal lactobacilli around weaning determined by means of 16S rRNA gene amplification and denaturing gradient gel electrophoresis
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DOI：
10.1111/j.1574-6941.2007.00317.x
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发表时间：
2007-07-01
影响因子：
4.2
通讯作者：
Souffrant, Wolfgang Bernhard
中科院分区：
文献类型：
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作者：
Janczyk, Pawel;Pieper, Robert;Souffrant, Wolfgang Bernhard
本研究以16S rRNA基因扩增和变性梯度凝胶电泳法(DGGE)为基础，对断奶前后猪肠道乳杆菌种群的变化进行了全面的研究。从断奶仔猪(28日龄)、1日龄、2日龄、5日龄和11日龄回肠内容物中提取DNA。扩增产物(16S rRNA基因的V2-V3片段)用DGGE分离。再扩增后切下优势条带并测序。断奶后1天和2天出现一条与唾液乳杆菌相对应的条带，而皱纹乳杆菌仅在断奶后1天和11天出现条带。温氏乳杆菌在所有动物中的优势条带最多。带数从断奶时的13+/-3条减少到断奶后5天的9+/-1条，但物种丰富度在断奶后11天有所恢复。采样日之间的剖面相似性在第1天和第2天(>；91%)较高，而在第5天和第11天(<；59%)较低。根据Shannon多样性指数，断奶后5日龄仔猪的多样性较低(0.83+/-0.076 vs.1.02+/-0.127，P=0.042)，但到11日龄时已恢复到断奶前水平。组特异性DGGE的应用表明，断奶后猪回肠内的乳杆菌群落发生了戏剧性的、部分可逆的变化。
Our study aimed to provide a comprehensive characterization of changes in porcine intestinal Lactobacillus populations around the time of weaning based on 16S rRNA gene amplification and denaturing gradient gel electrophoresis (DGGE). DNA was extracted from the ileal contents of piglets at weaning (28 days of age) and after 1, 2, 5 and 11 days. PCR amplicons (V2-V3 fragments of 16S rRNA genes) were separated using DGGE. Predominant bands were excised and sequenced after reamplification. A band corresponding to Lactobacillus salivarius was present 1 and 2 days post-weaning (pw), while Lactobacillus crispatus was detected only 1 and 11 days pw. Lactobacillus sobrius gave the most dominant band in all animals. The number of bands decreased from 13 +/- 3 at weaning to 9 +/- 1 at 5 days pw, but the species richness had recovered by 11 days pw. The similarity of profiles between sampling days was high for 1 and 2 days pw (> 91%), but was low for 5 and 11 days pw (< 59%). The diversity of the profiles was lower 5 days pw, based on the Shannon diversity index (0.83 +/- 0.076 vs. 1.02 +/- 0.127 at weaning, P=0.042), but had recovered to preweaning values by 11 days pw. The application of group-specific DGGE showed that the Lactobacillus community within the porcine ileum undergoes dramatic, partly reversible changes as a consequence of weaning.