Evaluating the microbial diversity of an in vitro model of the human large intestine by phylogenetic microarray analysis.
A high-density phylogenetic microarray targeting small subunit rRNA (SSU rRNA) sequences of over 1000 microbial phylotypes of the human gastrointestinal tract, the HITChip, was used to assess the impact of faecal inoculum preparation and operation conditions on an in vitro model of the human large intestine (TIM-2). This revealed that propagation of mixed faecal donations for the production of standardized inocula has only a limited effect on the microbiota composition, with slight changes observed mainly within the Firmicutes. Adversely, significant shifts in several major groups of intestinal microbiota were observed after inoculation of the in vitro model. Hierarchical cluster analysis was able to show that samples taken throughout the inoculum preparation grouped with microbiota profiles observed for faecal samples of healthy adults. In contrast, the TIM-2 microbiota was distinct. While members of the Bacteroidetes and some groups within the Bacilli were increased in TIM-2 microbiota, a strong reduction in the relative abundance of other microbial groups, including Bifidobacterium spp., Streptococcus spp., and Clostridium clusters IV and XIVa, was observed. The changes detected with the HITChip could be confirmed using denaturing gradient gel electrophoresis (DGGE) of SSU rRNA amplicons.
Rajilić-Stojanović
Mirjana
M
Laboratory of Microbiology, Wageningen University, Dreijenplein 10, 6703 HB Wageningen, The Netherlands.
Maathuis
Annet
A
Top Institute Food and Nutrition, PO Box 557, 6700 AN Wageningen, The Netherlands.|TNO Quality of Life, PO Box 360, 3700 AJ Zeist, The Netherlands.
Heilig
Hans G H J
HGHJ
Laboratory of Microbiology, Wageningen University, Dreijenplein 10, 6703 HB Wageningen, The Netherlands.
Venema
Koen
K
Top Institute Food and Nutrition, PO Box 557, 6700 AN Wageningen, The Netherlands.|TNO Quality of Life, PO Box 360, 3700 AJ Zeist, The Netherlands.
de Vos
Willem M
WM
Department of Veterinary Biosciences, PO Box 66, FN-00014, University of Helsinki, Finland.|Laboratory of Microbiology, Wageningen University, Dreijenplein 10, 6703 HB Wageningen, The Netherlands.
Smidt
Hauke
H
Top Institute Food and Nutrition, PO Box 557, 6700 AN Wageningen, The Netherlands.|Laboratory of Microbiology, Wageningen University, Dreijenplein 10, 6703 HB Wageningen, The Netherlands.
D001419:Bacteria / Q000235:genetics*; D001431:Bacteriological Techniques; D016000:Cluster Analysis; D004269:DNA, Bacterial / Q000235:genetics; D058645:Denaturing Gradient Gel Electrophoresis; D005243:Feces / Q000382:microbiology; D006801:Humans; D007420:Intestine, Large / Q000382:microbiology*; D054892:Metagenome; D008954:Models, Biological; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D010802:Phylogeny; D054680:Ribosome Subunits, Small, Bacterial / Q000235:genetics*
D004269:DNA, Bacterial