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对饲喂全株稻谷的猪的结肠内容物和粪便中细菌群落进行变性梯度凝胶电泳(DGGE)和16S核糖体DNA(rDNA)测序分析。

DGGE and 16S rDNA sequencing analysis of bacterial communities in colon content and feces of pigs fed whole crop rice.

作者信息

Wang Hai-Feng, Zhu Wei-Yun, Yao Wen, Liu Jian-Xin

机构信息

MOE Key Laboratory of Molecular Animal Nutrition, Institute of Dairy Science, Zhejiang University, Hangzhou 310029, PR China.

出版信息

Anaerobe. 2007 Jun-Aug;13(3-4):127-33. doi: 10.1016/j.anaerobe.2007.03.001. Epub 2007 Mar 12.

Abstract

The effect of feeding whole crop rice (WCR) to growing-finishing pigs at three levels 0 (Control), 10% and 20% on bacterial communities in colon content and feces was analyzed using 16S rDNA-based techniques. Amplicons of the V6-V8 variable regions of bacterial 16S rDNA were analyzed by denaturing gradient gel electrophoresis (DGGE), cloning and sequencing. The total number of DGGE bands and Shannon index of diversity for feces samples were higher in the pigs fed WCR-containing diets compared with the control, while a decrease trend was observed in these two parameters for colon content samples with the inclusion of WCR in the diets, although statistical differences were not significant. In general, the intestinal bacterial communities were prone to form the cluster for pig fed the same diet. Feeding of WCR induced the presence of special DGGE band with the sequence showing 99% similarity to that of Lactobacillus reuteri (DSM 20016T). The sequences of seven amplicons in total nine clones showed less than 97% similarity with those of previously identified or unidentified bacteria, suggesting that most bacteria in gastrointestinal tracts have not been cultured or identified. The results suggest that the diet containing WCR did not affect the major groups of bacteria, but stimulated the growth of L. reuteri-like species.

摘要

采用基于16S rDNA的技术,分析了以0(对照)、10%和20%三个水平的全株水稻(WCR)饲喂生长育肥猪对结肠内容物和粪便中细菌群落的影响。通过变性梯度凝胶电泳(DGGE)、克隆和测序分析细菌16S rDNA V6 - V8可变区的扩增子。与对照组相比,饲喂含WCR日粮的猪粪便样本的DGGE条带总数和香农多样性指数更高,而日粮中添加WCR时,结肠内容物样本的这两个参数呈下降趋势,尽管差异无统计学意义。总体而言,饲喂相同日粮的猪肠道细菌群落易于形成聚类。饲喂WCR诱导出现了一条特殊的DGGE条带,其序列与罗伊氏乳杆菌(DSM 20016T)的序列相似度为99%。在总共9个克隆中,有7个扩增子的序列与先前已鉴定或未鉴定的细菌序列相似度低于97%,这表明胃肠道中的大多数细菌尚未被培养或鉴定。结果表明,含WCR的日粮不影响主要细菌菌群,但刺激了类罗伊氏乳杆菌的生长。

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