Sarma-Rupavtarm Ramahi B, Ge Zhongming, Schauer David B, Fox James G, Polz Martin F
Department of Civil and Environmental Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA.
Appl Environ Microbiol. 2004 May;70(5):2791-800. doi: 10.1128/AEM.70.5.2791-2800.2004.
The overall complexity of the microbial communities in the gastrointestinal (GI) tracts of mammals has hindered observations of dynamics and interactions of individual bacterial populations. However, such information is crucial for understanding the diverse disease-causing and protective roles that gut microbiota play in their hosts. Here, we determine the spatial distribution, interanimal variation, and persistence of bacteria in the most complex defined-flora (gnotobiotic) model system to date, viz., mice colonized with the eight strains of the altered Schaedler flora (ASF). Quantitative PCR protocols based on the 16S rRNA sequence of each ASF strain were developed and optimized to specifically detect as few as 10 copies of each target. Total numbers of the ASF strains were determined in the different regions of the GI tracts of three C.B-17 SCID mice. Individual strain abundance was dependent on oxygen sensitivity, with microaerotolerant Lactobacillus murinus ASF361 present at 10(5) to 10(7) cells/g of tissue in the upper GI tract and obligate anaerobic ASF strains being predominant in the cecal and colonic flora at 10(8) to 10(10) cells/g of tissue. The variation between the three mice was small for most ASF strains, except for Clostridium sp. strain ASF502 and Bacteroides sp. strain ASF519 in the cecum. A comparison of the relative distribution of the ASF strains in feces and the colon indicated large differences, suggesting that fecal bacterial levels may provide a poor approximation of colonic bacterial levels. All ASF strains were detected by PCR in the feces of C57BL/6 restricted flora mice, which had been maintained in an isolator without sterile food, water, or bedding for several generations, providing evidence for the stability of these strains in the face of potential competition by bacteria introduced into the gut.
哺乳动物胃肠道(GI)中微生物群落的整体复杂性阻碍了对单个细菌种群动态和相互作用的观察。然而,这些信息对于理解肠道微生物群在其宿主中发挥的多种致病和保护作用至关重要。在这里,我们在迄今为止最复杂的特定菌群(悉生生物)模型系统中,即定殖有八株改变的 Schaedler 菌群(ASF)的小鼠中,确定了细菌的空间分布、个体间差异和持久性。基于每个 ASF 菌株的 16S rRNA 序列开发并优化了定量 PCR 方案,以特异性检测低至每个靶标的 10 个拷贝。在三只 C.B-17 SCID 小鼠的胃肠道不同区域测定了 ASF 菌株的总数。个体菌株丰度取决于对氧气的敏感性,微需氧的鼠李糖乳杆菌 ASF361 在上消化道中的含量为每克组织 10⁵至 10⁷个细胞,而专性厌氧的 ASF 菌株在盲肠和结肠菌群中占主导地位,每克组织含量为 10⁸至 10¹⁰个细胞。除了盲肠中的梭菌属菌株 ASF502 和拟杆菌属菌株 ASF519 外,大多数 ASF 菌株在三只小鼠之间的差异很小。ASF 菌株在粪便和结肠中的相对分布比较表明存在很大差异,这表明粪便细菌水平可能无法很好地反映结肠细菌水平。在几代以来一直饲养在没有无菌食物、水或垫料的隔离器中的 C57BL/6 限菌小鼠的粪便中,通过 PCR 检测到了所有 ASF 菌株,这为这些菌株在面对引入肠道的细菌的潜在竞争时的稳定性提供了证据。