The PBS medium contained NaCl at 8 g l−1, KCl at 0.2 g l−1, Na2HPO4 at 1.15 g l−1, KH2PO4 at 0.2 g l−1, L-Cysteine at 0.05%, adjusted to pH 7.3). The BCM medium contained peptone water at 2 g l−1, yeast extract at 2 g l−1, NaCl at 0.1 g l−1, K2HPO4 at 40 mg l−1, KH2PO4 at 40 mg l−1, MgSO4·7H2O at 10 mg l−1, CaCl2﹒6H2O at 10 mg l−1, NaHCO3 at 2 g l−1, L-cysteine at 0.05%, bile salts at 0.5 g l−1, vitamin K at 10 μl l−1, Tween 80 at 2 ml l−1 and hemin at 5 mg l−1, adjusted to pH 7.457 (link).
A fresh fecal sample was collected from a healthy woman, age 27, who had no known metabolic or gastrointestinal diseases and had taken no antibiotics or prebiotics for three months prior to the study. Written informed consent was obtained from this donor. The 10% (w/v) fecal slurry was prepared by diluting the fecal sample in sterile PBS medium, thoroughly suspended57 (link), and placed into an anaerobic chamber (H2:CO2:N2, 10:10:80, Whitley DG500 anaerobic work station, Don Whitley Scientific, West Yorkshire, UK) within 30 min after collection. After being filtered through two layers of gauze, the fecal solution was inoculated into the BCM and PBS batch culture systems.
The total volume of each culture system was 20 ml. The PBS system was started with 5% fecal inoculum added with 10 ml of 10% fecal slurry, and the BCM system was started with 1% fecal inoculum added with 2 ml of 10% fecal slurry. Negative control cultures consisted of culture medium and inoculum but no prebiotic substrate. Prebiotic cultures consisted of culture medium, inoculum and a prebiotic formula (2.5%, w/v), which was a mixture of galactooligosaccharide and guar gum at a ratio of 1:1 (w/w). Cultures of negative control and prebiotic formula were performed in the PBS and BCM systems, respectively, in an anaerobic chamber at 37 °C without stirring, and samples were dynamically collected at 0, 6, 24 and 72 h in both systems and both prebiotic and negative control cultures. This study was approved by the School of Shanghai Jiao Tong University Ethics Committee Biomedical Project (document no. 2014-018), and all experiments were performed in accordance with the relevant guidelines and regulations.
A fresh fecal sample was collected from a healthy woman, age 27, who had no known metabolic or gastrointestinal diseases and had taken no antibiotics or prebiotics for three months prior to the study. Written informed consent was obtained from this donor. The 10% (w/v) fecal slurry was prepared by diluting the fecal sample in sterile PBS medium, thoroughly suspended57 (link), and placed into an anaerobic chamber (H2:CO2:N2, 10:10:80, Whitley DG500 anaerobic work station, Don Whitley Scientific, West Yorkshire, UK) within 30 min after collection. After being filtered through two layers of gauze, the fecal solution was inoculated into the BCM and PBS batch culture systems.
The total volume of each culture system was 20 ml. The PBS system was started with 5% fecal inoculum added with 10 ml of 10% fecal slurry, and the BCM system was started with 1% fecal inoculum added with 2 ml of 10% fecal slurry. Negative control cultures consisted of culture medium and inoculum but no prebiotic substrate. Prebiotic cultures consisted of culture medium, inoculum and a prebiotic formula (2.5%, w/v), which was a mixture of galactooligosaccharide and guar gum at a ratio of 1:1 (w/w). Cultures of negative control and prebiotic formula were performed in the PBS and BCM systems, respectively, in an anaerobic chamber at 37 °C without stirring, and samples were dynamically collected at 0, 6, 24 and 72 h in both systems and both prebiotic and negative control cultures. This study was approved by the School of Shanghai Jiao Tong University Ethics Committee Biomedical Project (document no. 2014-018), and all experiments were performed in accordance with the relevant guidelines and regulations.
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