Title:
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Development of a system for modelling of the human large intestine
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Laboratory model
This project covers the design and production of a new continuous culture system for
the modelling of the human large intestine. A single stage tubular model of the human
ascending colon was produced through the continued adaptation of preliminary
models. This single- stage model consisted of a membrane based fermenter used in
conjunction with an aqueous polyethylene glycol solution for water and metabolite
removal and the pH control of the fermentation media. The design also incorporated a
module for sampling media during the course of the fermentation. This single stage
model was characterised through fermentations using a faecal inoculum. Properties
monitored were the ability to control media pH indirectly through the transfer of
solutes across a membrane, the degree of water removal through the membrane and
the degree of substrate utilisation in the system. The ability of the fermenter to
maintain the growth of bacterial species present in the gut, at a flow rate analogous to
that through the human colon was also monitored. The membrane system was found
to facilitate the control of the pH in the fermentation media. The removal of water and
short chain fatty acids from fermentation media was also observed. Large and stable
numbers of colonic bacteria were maintainable in the fermenter.
The single- stage model provided the basis for a three- stage tubular model of the
large intestine which was produced and analysed by means of inoculation with a
faecal sample and monitoring its performance during a fermentation.
The 3- stage tubular system was found to give a good representation of conditions in
the human large intestine. Numbers of each bacterial group studied were consistently higher in the membrane system than in an existing gut model system (Macfarlane et
al. 1998). This suggested that the tubular membrane model provided a better
environment for bacterial growth and a better representation of the actual colonic
micro flora. The new system was found to be capable of maintaining total bacterial
numbers of 5.0 ± 4.8 xl010 bacterial ml in the first fermenter section. Concentrations
of short chain fatty acids throughout the system were found to be similar to those
associated with the corresponding regions of the human colon. The membrane system
was capable of controlling the pH throughout the fermentation media and providing a
degree of short chain fatty acid and water removal. The model was capable of
removing 64% of incoming media across the membranes, the majority of which can
be assumed to be water. This provided a good representation of the degree of water
removal that occurs in the human colon itself.
Mathematical Model
To supplement the practical model, mixing was characterised in a tubular fermenter
with intermediate flow regime. To this aim, diffusion-convection and diffusion
models were solved numerically to provide the theoretical residence time distribution
,in tubular fermenters, in intermediate flow regimes. The diffusion-convection model
was validated using the experimental results obtained from step tracer experiments in
a tubular vessel at different flow rates and was shown to correlate well with
experimental data. The variance ratio between the two models was calculated
allowing its use as a correction factor. This allows the use of the diffusion model for
the characterisation of mixing in tubular fermenters that are associated with low
Reynolds numbers, such as the new gut model system.
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