In vitro maintenance of a human proximal colon microbiota using the continuous fermentation system P-ECSIM |
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Authors: | David Feria-Gervasio Sylvain Denis Monique Alric Jean-François Brugère |
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Institution: | 1.Conception ingénierie et développement de l’aliment et du médicament,Clermont Université, Université d’Auvergne, ERT 18-CIDAM,Clermont-Ferrand,France |
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Abstract: | Ethical and technical difficulties for in vivo studies on gut microbiotas argue for the development of alternative in vitro
models: here, we describe a system simulating the proximal part of a human colon both nutritionally and physico-chemically
with a procedure aimed to limit experimental variations over the time (Proximal Environmental Control System For Intestinal
Microbiota—P-ECSIM). The continuous culture system P-ECSIM is first inoculated by a −20°C glycerol stock established from
the batch culture of a stool-inoculated medium. The anaerobic atmosphere is self-maintained by the gases produced in the ordinary
metabolism of fermentations. The monitoring of metabolic activities and microbial constitutions indicates that different steady
states are obtained according to the dilution rate. Finally, the glycerol conservation of the batch culture-derived inoculum
gives a similar differential response between the two dilution rates (D = 0.08 h−1 and D = 0.04 h−1) after a 1-year storage time as well for their metabolism and constitution in steady states, but with a lower abundance.
Molecular fingerprints of the microbiota reveal however alterations over the time. Further efforts are needed concerning the
preservation of standardized inoculums in order to improve the process for intra- and inter-lab comparison. Combined with
appropriate analytical techniques, this system provides an efficient alternative means of studying functionally human microbiota
in its constitution, metabolism and adaptation to environmental changes, particularly nutritional. |
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