engleski

System for maintenance of human gut microbiota using an in vitro large intestine model

The aim of the study is to create in vitro system based on bioreactor that is able to simulate environmental conditions of three different parts of the human large intestine. The first stage of the study is focused on defining the critical process parameters during repeated batch cultivation in order to maintain microbial communities similar to donor microbiota, primary from healthy obese subjects. When those conditions are met in second stage system will be used for testing the effects of prebiotics (Nutriose) and probiotics (Lactobacillus paraplantarum) on the composition of human gut microbiota in obese subjects. For the first stage of the experiment composition of ileal efflux media was defined based on data from the literature. Selected medium simulates the undigested part of food and its nutrient characteristic of a western diet. After the adaptation period of 4 h fresh fecal sample from the obese donor was homogenized in dialysate and used as an inoculum. The experiment was conducted for 78 h. The medium replacement was carried out daily for three days. Samples were taken every 8 h and stored at -80°C. Isolated DNA form the samples will be used for determining biodiversity based on 16S rRNA gene sequencing. Based on results of temporal microbial biodiversity normal variance of the microbiota on the phylum level will be defined.

human gut microbiota ; in vitro gut model ; genomic analysis ; probiotic ; prebiotic ; obesity

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