Zoetendal Erwin G, von Wright Atte, Vilpponen-Salmela Terttu, Ben-Amor Kaouther, Akkermans Antoon D L, de Vos Willem M
Wageningen Centre for Food Sciences, Laboratory of Microbiology, Hesselink van Suchtelenweg 4, 6703 CT Wageningen, The Netherlands.
Appl Environ Microbiol. 2002 Jul;68(7):3401-7. doi: 10.1128/AEM.68.7.3401-3407.2002.
The human gastrointestinal (GI) tract harbors a complex community of bacterial cells in the mucosa, lumen, and feces. Since most attention has been focused on bacteria present in feces, knowledge about the mucosa-associated bacterial communities in different parts of the colon is limited. In this study, the bacterial communities in feces and biopsy samples from the ascending, transverse, and descending colons of 10 individuals were analyzed by using a 16S rRNA approach. Flow cytometric analysis indicated that 10(5) to 10(6) bacteria were present in the biopsy samples. To visualize the diversity of the predominant and the Lactobacillus group community, denaturing gradient gel electrophoresis (DGGE) analysis of 16S rRNA gene amplicons was performed. DGGE analysis and similarity index comparisons demonstrated that the predominant mucosa-associated bacterial community was host specific and uniformly distributed along the colon but significantly different from the fecal community (P < 0.01). The Lactobacillus group-specific profiles were less complex than the profiles reflecting the predominant community. For 6 of the 10 individuals the community of Lactobacillus-like bacteria in the biopsy samples was similar to that in the feces. Amplicons having 99% sequence similarity to the 16S ribosomal DNA of Lactobacillus gasseri were detected in the biopsy samples of nine individuals. No significant differences were observed between healthy and diseased individuals. The observed host-specific DGGE profiles of the mucosa-associated bacterial community in the colon support the hypothesis that host-related factors are involved in the determination of the GI tract microbial community.
人类胃肠道(GI)的黏膜、管腔和粪便中栖息着复杂的细菌群落。由于大多数研究都聚焦于粪便中的细菌,因此关于结肠不同部位黏膜相关细菌群落的了解有限。在本研究中,采用16S rRNA方法分析了10名个体粪便以及升结肠、横结肠和降结肠活检样本中的细菌群落。流式细胞术分析表明,活检样本中存在10⁵至10⁶个细菌。为了直观呈现优势菌群落和乳酸杆菌菌群的多样性,对16S rRNA基因扩增子进行了变性梯度凝胶电泳(DGGE)分析。DGGE分析和相似性指数比较表明,主要的黏膜相关细菌群落具有宿主特异性,沿结肠均匀分布,但与粪便群落显著不同(P < 0.01)。乳酸杆菌菌群特异性图谱不如反映优势菌群的图谱复杂。10名个体中有6名的活检样本中类乳酸杆菌细菌群落与粪便中的相似。在9名个体的活检样本中检测到与加氏乳杆菌16S核糖体DNA序列相似性达99%的扩增子。健康个体和患病个体之间未观察到显著差异。在结肠中观察到的黏膜相关细菌群落的宿主特异性DGGE图谱支持了宿主相关因素参与胃肠道微生物群落决定的假说。