采用聚合酶链式反应-变性梯度凝胶电泳技术分析小熊猫胃肠道菌群的多样性

Bacteria Diversity in Gastrointestinal Tract of Ailurus fulgens Analyzed by Polymerase Chain Reaction-Denaturing Gradient Gel Electrophoresis Technology

本试验旨在研究小熊猫胃肠道菌群多样性。采用聚合酶链式反应(PCR)-变性梯度凝胶电泳(DGGE)技术结合条带的克隆测序、聚类分析和主成分分析(PCA)检测小熊猫胃肠道菌群结构及多样性。结果表明:1)PCR-DGGE图谱显示小熊猫胃肠道中有大量菌群,且不同部位的菌群结构存在一定的差异,相邻肠段菌群结构具有一定的相似性。结肠和粪便样品的菌群多样性较高,其次为胃和直肠样品,而空肠和回肠样品菌群多样性较低。2)小熊猫胃肠道菌群PCR-DGGE图谱中测序的条带大多数归为厚壁菌门(Firmicutes)、拟杆菌门(Bacteroides)、变形菌门(Proteobacteria)和疣微菌门(Verrucomicrobia),共性条带主要是未培养拟杆菌门细菌(uncultured Bacteroidetes bacterium)、粪肠球菌(Enterococcus faecalis)、未培养梭状芽孢杆菌(uncultured Clostridium sp.)、乳酸乳球菌(Lactococcus lactis)和食窦魏斯氏菌(Weissella cibaria),其中厚壁菌门为优势菌群;特异性条带主要是丛毛单胞菌(Comamonas sp.)、梭菌属(Clostridium sp.)和Akkermansia。由此可见,小熊猫胃肠道中栖息着大量菌群,且其多样性按照胃肠道由前至后的顺序呈现高-低-高的趋势。

This study aimed to evaluate the bacterial diversity in the gastrointestinal tract （GIT） of Ailurus fulgens. The polymerase chain reaction （PCR）-denaturing gradient gel electrophoresis （DGGE） technology combined with the cloning sequencing, cluster analysis and principal component analysis （PCA） of bands were used to detect the bacterial composition and diversity. The results showed as follows：1） numerous bacteria were obtained from the GIT of Ailurus fulgens by using the PCR-DGGE profiles. There was a difference in each intestinal segment. However, there were some similarities between the adjacent intestinal segment. Samples from the colon and faeces had the higher bacterial diversity, the secondly for samples from stomach and rectum, and the samples from the jejunum and ileum had the lower bacterial diversity. 2） The sequencing results of PCR-DGGE profiles of bacteria from gastrointestinal tract of Ailurus fulgens showed that most of the bacteria belonged to Firmicutes, Bacteroides, Proteobacteria and Verrucomicrobia. The common bands were mainly inclued uncultured Bacteroidetes bacterium, Enterococcus faecalis, uncultured Clostridium sp., Lactococcus lactis and Weissella cibaria, and Firmicutes was the dominant bacterial. The specific bands were Comamonas sp., Clostridium sp. and Akkermansia. In summary, a large amounts of bacteria exist in the GIT of Ailurus fulgens, and the diversity of bacteria shows a tendency of high-low-high according to the GIT from front to back.