Maturation of the murine cecal microbiota as revealed by terminal restriction fragment length polymorphism and 16S rRNA gene clone libraries

Abstract

The maturation of murine cecal microbiota was determined by terminal restriction fragment polymorphism (T-RFLP) and 16S rRNA gene clone libraries. Cecal microbiota in specific pathogen free (SPF) mice aged four to 10 weeks were collected. The cluster of samples in 4-week-old mice was different from those of other ages based on T-RFLP profiles. The majority of clones obtained in this study belonged to the Clostridium coccoides (C. coccoides) group, the Bacteroides group or the Lactobacillus group. Phylogenetic analysis showed characteristic clusters composed of new operational taxonomic unit (OTU) of the C. coccoides and Bacteroides groups. The existence of a large number of yet unidentified bacteria inhabiting the murine cecum was demonstrated by 16S rRNA gene clone libraries. T-RFLP analysis data were more complex and more sensitive than the patterns generated by computer simulation of 16S rRNA gene clone library analysis data. T-RFLP revealed development with maturation of cecal microbiota including unidentified bacteria of SPF mice.

Nod2 is essential for temporal development of intestinal microbial communities

“…A detailed view into the colonisation process during mouse development highlights that microbial composition is not only influenced by genotype, but also by stage of development. Previous studies based on fingerprinting approaches found that the caecal microbiota in SPF mice changes drastically with age, but stabilises after 4 weeks 45. In our study based on 16S rRNA gene clone libraries, the microbial community continued to change after week 4b, fluctuating in the proportions of the three most abundant phyla ( Firmicutes , Bacteroidetes and Proteobacteria ; figure 1B) until 10 weeks of age, at which point stable Firmicutes -dominated communities became apparent.…”

The Human Gut Microbiome and Body Metabolism: Implications for Obesity and Diabetes

“…Sequencing of 2 primary targets within bacterial 16S rRNA genes yielded valuable compositional data pertaining to the human fecal microbiome of 242 healthy adults ( 6, 7 ) . In the Human Microbiome Project, 18 different body sites were sampled and sequenced.…”

“…Previously published studies demonstrated the variation in composition of the gut microbiome among locations within the gastrointestinal tract in different mammalian species. For example, 16S rRNA gene sequencing has been deployed to study the maturation of murine cecal microbiota, and these studies demonstrated the existence of a large number of yet-unidentified bacteria that inhabit the mammalian intestine ( 6 ) . Such sequencing strategies, which are culture independent, are essential for determining bacterial composition of the microbiome and its relative stability and diversity over time.…”

Change of Intestinal Microbiota with Elemental Diet and Its Impact on Therapeutic Effects in a Murine Model of Chronic Colitis

“…Polymerase chain reaction (PCR) amplification was performed as described by Kibe et al [21]. Two pairs of primers were used: 27f (5 0 -AGAGTTTGATCCTGGCT-CAG-3 0 ) and 1492r (5 0 -GGTTACCTTGTTACGACTT-3 0 ) as the universal primer, and 27f and Lab-677r (5 0 -CAC-CGCTACACATGGAG-3 0 ) as the specific primer set for lactic acid bacteria (LAB) designed by Heilig et al [22].…”