Di Cagno Raffaella, Rizzello Carlo G, Gagliardi Francesca, Ricciuti Patrizia, Ndagijimana Maurice, Francavilla Ruggiero, Guerzoni M Elisabetta, Crecchio Carmine, Gobbetti Marco, De Angelis Maria
Department of Plant Protection and Applied Microbiology, University of Bari, Italy.
Appl Environ Microbiol. 2009 Jun;75(12):3963-71. doi: 10.1128/AEM.02793-08. Epub 2009 Apr 17.
This study aimed at investigating the fecal microbiotas of children with celiac disease (CD) before (U-CD) and after (T-CD) they were fed a gluten-free diet and of healthy children (HC). Brothers or sisters of T-CD were enrolled as HC. Each group consisted of seven children. PCR-denaturing gradient gel electrophoresis (DGGE) analysis with V3 universal primers revealed a unique profile for each fecal sample. PCR-DGGE analysis with group- or genus-specific 16S rRNA gene primers showed that the Lactobacillus community of U-CD changed significantly, while the diversity of the Lactobacillus community of T-CD was quite comparable to that of HC. Compared to HC, the ratio of cultivable lactic acid bacteria and Bifidobacterium to Bacteroides and enterobacteria was lower in T-CD and even lower in U-CD. The percentages of strains identified as lactobacilli differed as follows: HC (ca. 38%) > T-CD (ca. 17%) > U-CD (ca. 10%). Lactobacillus brevis, Lactobacillus rossiae, and Lactobacillus pentosus were identified only in fecal samples from T-CD and HC. Lactobacillus fermentum, Lactobacillus delbrueckii subsp. bulgaricus, and Lactobacillus gasseri were identified only in several fecal samples from HC. Compared to HC, the composition of Bifidobacterium species of T-CD varied, and it varied even more for U-CD. Forty-seven volatile organic compounds (VOCs) belonging to different chemical classes were identified using gas-chromatography mass spectrometry-solid-phase microextraction analysis. The median concentrations varied markedly for HC, T-CD, and U-CD. Overall, the r(2) values for VOC data for brothers and sisters were equal to or lower than those for unrelated HC and T-CD. This study shows the effect of CD pathology on the fecal microbiotas of children.
本研究旨在调查患有乳糜泻(CD)的儿童在食用无麸质饮食之前(U-CD)和之后(T-CD)以及健康儿童(HC)的粪便微生物群。T-CD的兄弟姐妹被纳入作为HC。每组由7名儿童组成。使用V3通用引物的PCR-变性梯度凝胶电泳(DGGE)分析揭示了每个粪便样本的独特图谱。使用组特异性或属特异性16S rRNA基因引物的PCR-DGGE分析表明,U-CD的乳酸杆菌群落发生了显著变化,而T-CD的乳酸杆菌群落多样性与HC相当。与HC相比,T-CD中可培养乳酸菌和双歧杆菌与拟杆菌和肠杆菌的比例较低,U-CD中更低。鉴定为乳酸杆菌的菌株百分比差异如下:HC(约38%)>T-CD(约17%)>U-CD(约10%)。短乳杆菌、罗斯氏乳杆菌和戊糖乳杆菌仅在T-CD和HC的粪便样本中鉴定到。发酵乳杆菌、德氏乳杆菌保加利亚亚种和加氏乳杆菌仅在HC的几个粪便样本中鉴定到。与HC相比,T-CD的双歧杆菌种类组成有所不同,U-CD的变化更大。使用气相色谱-质谱-固相微萃取分析鉴定了47种属于不同化学类别的挥发性有机化合物(VOC)。HC、T-CD和U-CD的中位浓度差异显著。总体而言,兄弟姐妹的VOC数据的r(2)值等于或低于无关的HC和T-CD。本研究显示了CD病理对儿童粪便微生物群的影响。