Swidsinski A, Loening-Baucke V, Kirsch S, Doerffel Y
Laboratory for Molecular Genetics, Polymicrobial Infections and Bacterial Biofilms and Department of Medicine, Section of Gastroenterology, Charité Universitätsmedizin Berlin, Berlin, Germany.
Gastroenterol Clin Biol. 2010 Sep;34 Suppl 1:S79-92. doi: 10.1016/S0399-8320(10)70025-7.
The colonic content can be compared to a spatially structured high output bioreactor composed of three functionally different regions: a separating mucus layer, a germinal stock area, and a central fermenting area. The stool mirrors this structure and can be used for diagnosis in health and disease. In a first part, we introduce a novel method based on fluorescence in situ hybridization (FISH) of sections of punched-out stool cylinders, which allows quantitatively monitor microbiota in the mucus, the germinal stock and the central fermenting areas. in a second part, we demonstrate the practical implementation of this method, describing the biostructure of stool microbiota in healthy subjects and patients with chronic idiopathic diarrhea treated with Saccharomyces boulardii. Punched stool cylinders from 20 patients with chronic idiopathic diarrhea and 20 healthy controls were investigated using fluorescence in situ hybridization. Seventy-three bacterial groups were evaluated. Fluctuations in assembly of 11 constitutive bacterial groups were monitored weekly for 3 weeks prior to, 3 weeks during, and 3 weeks after oral Saccharomyces boulardii supplementation. Typical findings in healthy subjects were a 5-60 μm mucus separating layer; homogeneous distribution and fluorescence, high concentrations (>10 × 10(10) bacterial/mL) of the three habitual bacterial groups: Bacteroides, Roseburia and Faecalibacterium prausnitzii; and low concentrations of the occasional bacterial groups. The diarrhea could be described in terms of increased separating effort, purging, decontamination, bacterial substitution. Typical findings in diarrhea were: increased thickness of the protective mucus layer, its incorporation in the stool, absolute reduction in concentrations of the habitual bacterial groups, suppression of bacterial metabolism in the central fermenting area (hybridization silence), stratification of the stool structure by watery ingredients, and substitutive increase in the concentrations of occasional bacterial groups. The microbial and clinical symptoms of diarrhea were reversible with Saccharomyces boulardii therapy. The structure-functional analysis of stool microbiota allows to quantitatively monitor colonic malfunction and its response to therapy. Saccharomyces boulardii significantly improves the stool biostructure in patients with chronic idiopathic diarrhea and has no influence on the stool microbiota in healthy subjects.
结肠内容物可被比作一个空间结构的高产出生物反应器，它由三个功能不同的区域组成：一个分隔性的黏液层、一个生发菌库区域和一个中央发酵区域。粪便反映了这种结构，可用于健康和疾病的诊断。在第一部分中，我们介绍了一种基于对打孔粪便圆柱体切片进行荧光原位杂交（FISH）的新方法，该方法可定量监测黏液、生发菌库和中央发酵区域中的微生物群。在第二部分中，我们展示了该方法的实际应用，描述了健康受试者以及接受布拉氏酵母菌治疗的慢性特发性腹泻患者粪便微生物群的生物结构。使用荧光原位杂交技术对20例慢性特发性腹泻患者和20例健康对照的打孔粪便圆柱体进行了研究。评估了73个细菌群。在口服布拉氏酵母菌补充剂之前3周、期间3周和之后3周，每周监测11个组成性细菌群组装的波动情况。健康受试者的典型表现为：一个5 - 60μm的黏液分隔层；均匀分布和荧光，三种常见细菌群（拟杆菌属、罗斯氏菌属和普拉梭菌）的高浓度（>10×10¹⁰细菌/mL）；以及偶尔出现的细菌群的低浓度。腹泻可通过分离作用增强、清除、去污、细菌替代来描述。腹泻的典型表现为：保护性黏液层厚度增加，其融入粪便中，常见细菌群浓度绝对降低，中央发酵区域细菌代谢受抑制（杂交沉默），粪便结构因水性成分分层，以及偶尔出现的细菌群浓度替代性增加。腹泻的微生物和临床症状通过布拉氏酵母菌治疗是可逆的。粪便微生物群的结构 - 功能分析有助于定量监测结肠功能障碍及其对治疗的反应。布拉氏酵母菌显著改善慢性特发性腹泻患者的粪便生物结构，对健康受试者的粪便微生物群无影响。
