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Journal of Biomedicine and Biotechnology
Volume 2011 (2011), Article ID 838040, 10 pages
http://dx.doi.org/10.1155/2011/838040
Research Article

In Vitro Culture Conditions for Maintaining a Complex Population of Human Gastrointestinal Tract Microbiota

Division of Microbiology, National Center for Toxicological Research/U.S. FDA, Jefferson, AR 72079, USA

Received 13 January 2011; Revised 13 April 2011; Accepted 27 May 2011

Academic Editor: Eric C. Martens

Copyright © 2011 Bong-Soo Kim et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Supplementary Material

Figure S1. The growth of intestinal microbiota was analyzed on different media with 1% fecal inoculum. (a) Growth curves were constructed by OD600, and (b) 16S rRNA gene copies of total bacteria were quantified by real-time PCR. Growth of intestinal microbiota in low carbohydrate medium (green), high carbohydrate medium (red), and BHI (blue).

Figure S2. Decolorization of gentian violet by fecal bacterial culture after 18 hours. Fecal inocula from three individuals were incubated with 6.5 µM gentian violet for 18 hours. Gentian violet is reductively decolorized and decolorization indicates the metabolic activity of cultured bacteria. The control sample (C) incubated gentian violet to LCM for 18 hours without fecal inoculum. The fecal inoculum from individual A (2) incubated for 18 hours with gentian violet, individual B (3) incubated with gentian violet, and individual C (5) incubated with gentian violet.

Figure S3. The DGGE profiles of each concentration inoculum (0.1, 0.5, 1, 2, 3%) from individual D at different time point were generated from amplified V3 16S rRNA gene of genomic DNA. M denotes marker lanes, F denotes original fecal microbiota, and each number above lane is culture time (hour).

Figure S4. The growth of different fecal inoculum concentration from individual D was analyzed with spectrophotometer and quantitative real-time PCR. (a) Growth curves were constructed by ∆OD600, and (b) total bacterial 16S rRNA gene copies were compared.

Figure S5. The rarefaction curves for three individual samples of 0 hour and 18 hour. The numbers of observed phylotypes (OTUs) from samples were calculated by 97% similarity.

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