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. 2013 Jan 26;13:20.
doi: 10.1186/1471-230X-13-20.

Clostridium leptum group bacteria abundance and diversity in the fecal microbiota of patients with inflammatory bowel disease: a case-control study in India

Jayakanthan Kabeerdoss1, Vijayalakshmi Sankaran, Srinivasan Pugazhendhi, Balakrishnan S Ramakrishna
Affiliations
Free PMC article

Clostridium leptum group bacteria abundance and diversity in the fecal microbiota of patients with inflammatory bowel disease: a case-control study in India

Jayakanthan Kabeerdosset al. BMC Gastroenterol. .
Free PMC article

Abstract

Background:Alterations in the fecal bacterial flora occur in inflammatory bowel disease (IBD). We examined the abundance and diversity of Clostridium leptum group, an important group of carbohydrate-fermenting bacteria, in the feces of patients with IBD and compared them with healthy controls.

Methods:Seventeen healthy controls (HC), 20 patients with Crohn's disease (CD) and 22 patients with ulcerative colitis (UC) participated in the study. DNA extracted from fecal samples was amplified by PCR targeting 16S rRNA gene sequences specific to C. leptum group. The PCR product was subjected to temporal temperature gradient electrophoresis (TTGE) and the number and position of individual bands were noted and diversity was estimated. The identity of bands at different positions was confirmed by cloning and sequencing. Real time quantitative PCR with Mesa Green, targeted at specific 16S rRNA gene sequences, was used to quantitate C. leptum group and its most prominent constituent, Faecalibacterium prausnitzii.

Results:Twenty five different operational taxonomic units (OTUs, equivalent to species) were identified constituting the C. leptum group in these participants. Their sequences were deposited in GenBank [accession numbers GQ465348 to GQ465370]. OTU number was significantly reduced in CD (7.7 ± 3.7, mean ± SD) and UC (9.0 ± 3.0) compared to HC (11.9 ± 2.2) (P=0.0005). The Simpson D index of alpha diversity was not significantly different between the three groups. Total numbers of C. leptum group bacteria and F. prausnitzii were reduced in both CD and UC compared to HC (P=0.0036 and P<0.0001 respectively). Disease activity did not influence numbers of C. leptum or F. prausnitzii in patients with CD or UC.

Conclusion:C. leptum numbers and diversity were significantly reduced in both CD and UC suggesting that alterations noted were not specific to one disease. This could contribute to reduced short chain fatty acid production in IBD.

Figures

Figure 1
Figure 1
TTGE gel showing bands migrating at different positions.Each lane depicts fecal DNA amplified using the Clept primers from an individual participant. Details of the TTGE conditions are provided in the text.
Figure 2
Figure 2
Principal components analysis of fecal C. leptum group bacterial communities. C. leptum在健康对照组物种占优势(HC, blue) tend to cluster to the right side of the plot, while those species dominant in Crohn’s disease (CD, green) and ulcerative colitis (UC, turqoise) patients tend to cluster to the left of the plot.
Figure 3
Figure 3
Bray Curtis cluster analysis of fecal C. leptum group communities.Bacteria from Crohn’s disease (CD) and ulcerative colitis (UC) patients appeared to have higher degrees of dissimilarity to each other than bacteria from healthy controls (HC).
Figure 4
Figure 4
Number of C. leptum species in the feces of individuals in the three groups.Values shown are mean (SEM).
Figure 5
Figure 5
Abundance of C. leptum group bacteria (A) and F. prausnitzii (B).Bacterial abundance in HC, CD and UC, is expressed as mean (SEM) copy number of 16S rRNA gene/g feces.
Figure 6
Figure 6
Abundance of C. leptum group bacteria and F. prausnitzii in active and quiescent IBD.Values shown are mean (SEM) of 16S rRNA gene copy number per g feces in patients with CD and UC with quiescent and active disease.
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