Quantitative fluorescence in situ hybridization of Bifidobacterium spp. with genus-specific 16S rRNA-targeted probes and its application in fecal samples.
Petra S. Langendijk,Frits Schut,Gijsbert J. Jansen,Gerwin C. Raangs,Ger R. Kamphuis,Michael H. F. Wilkinson,Gjalt W. Welling +6 more
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Since the total culturable counts were only a fraction of the total microscopic counts, the contribution of bifidobacteria to the total intestinal microflora was overestimated by almost 10-fold when cultural methods were used as the sole method for enumeration.Abstract:
Three 16S rRNA hybridization probes were developed and tested for genus-specific detection of Bifidobacterium species in the human fecal flora. Variable regions V2, V4, and V8 of the 16S rRNA contained sequences unique to this genus and proved applicable as target sites for oligodeoxynucleotide probes. Determination of the genus specificity of the oligonucleotides was performed by whole-cell hybridization with fluorescein isothiocyanate-labelled probes. To this end, cells were fixed on glass slides, hybridized with the probes, and monitored by videomicroscopy. In combination with image analysis, this allowed quantification of the fluorescence per cell and objective evaluation of hybridization experiments. One of the probes developed was used to determine the population of Bifidobacterium spp. in human fecal samples. A comparison was made with results obtained by cultural methods for enumeration. Since both methods gave similar population estimates, it was concluded that all bifidobacteria in feces were culturable. However, since the total culturable counts were only a fraction of the total microscopic counts, the contribution of bifidobacteria to the total intestinal microflora was overestimated by almost 10-fold when cultural methods were used as the sole method for enumeration.read more
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Revised Estimates for the Number of Human and Bacteria Cells in the Body.
Ron Sender,Shai Fuchs,Ron Milo +2 more
TL;DR: This analysis updates the widely-cited 10:1 ratio, showing that the number of bacteria in the body is actually of the same order as the numberof human cells, and their total mass is about 0.2 kg.
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Development of the human infant intestinal microbiota.
Chana Palmer,Elisabeth M. Bik,Daniel B. DiGiulio,Daniel B. DiGiulio,David A. Relman,David A. Relman,Patrick O. Brown +6 more
TL;DR: A microarray is designed to detect and quantitate the small subunit ribosomal RNA (SSU rRNA) gene sequences of most currently recognized species and taxonomic groups of bacteria and suggested that incidental environmental exposures play a major role in determining the distinctive characteristics of the microbial community in each baby.
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Dietary modulation of the human colonic microbiota: updating the concept of prebiotics
TL;DR: The future use of prebiotics may allow species-level changes in the microbiota, an extrapolation into genera other than the bifidobacteria and lactobacilli, and allow preferential use in disease-prone areas of the body.
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Selective increases of bifidobacteria in gut microflora improve high-fat-diet-induced diabetes in mice through a mechanism associated with endotoxaemia.
Patrice D. Cani,Audrey M. Neyrinck,Francesca Fava,Claude Knauf,Rémy Burcelin,Kieran Tuohy,Glenn R. Gibson,Nathalie M. Delzenne,Nathalie M. Delzenne +8 more
TL;DR: The findings suggest that the gut microbiota contribute towards the pathophysiological regulation of endotoxaemia and set the tone of inflammation for occurrence of diabetes and/or obesity.
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Direct Analysis of Genes Encoding 16S rRNA from Complex Communities Reveals Many Novel Molecular Species within the Human Gut
Antonia Suau,Antonia Suau,Régis Bonnet,Malène Sutren,Jean Jacques Godon,Glenn R. Gibson,Matthew D. Collins,Joël Doré +7 more
TL;DR: The majority of generated rDNA sequences did not correspond to known organisms and clearly derived from hitherto unknown species within this human gut microflora, including Clostridium coccoides and Eubacterium rectale.
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