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. 2020 Jan 30;21(3):906.
doi: 10.3390/ijms21030906.

In Vitro Evaluation of Different Prebiotics on the Modulation of Gut Microbiota Composition and Function in Morbid Obese and Normal-Weight Subjects

Alicja M Nogacka  1   2 , Nuria Salazar  1   2 , Silvia Arboleya  1   2 , Patricia Ruas-Madiedo  1   3 , Leonardo Mancabelli  4 , Adolfo Suarez  2   5 , Ceferino Martinez-Faedo  6   7 , Marco Ventura  4 , Takumi Tochio  8 , Katsuaki Hirano  8 , Akihito Endo  9 , Clara G de Los Reyes-Gavilán  1   2 , Miguel Gueimonde  1   2
Affiliations

In Vitro Evaluation of Different Prebiotics on the Modulation of Gut Microbiota Composition and Function in Morbid Obese and Normal-Weight Subjects

Alicja M Nogacka et al. Int J Mol Sci. .

Abstract

The gut microbiota remains relatively stable during adulthood; however, certain intrinsic and environmental factors can lead to microbiota dysbiosis. Its restoration towards a healthy condition using best-suited prebiotics requires previous development of in vitro models for evaluating their functionality. Herein, we carried out fecal cultures with microbiota from healthy normal-weight and morbid obese adults. Cultures were supplemented with different inulin-type fructans (1-kestose, Actilight, P95, Synergy1 and Inulin) and a galactooligosaccharide. Their impact on the gut microbiota was assessed by monitoring gas production and evaluating changes in the microbiota composition (qPCR and 16S rRNA gene profiling) and metabolic activity (gas chromatography). Additionally, the effect on the bifidobacterial species was assessed (ITS-sequencing). Moreover, the functionality of the microbiota before and after prebiotic-modulation was determined in an in vitro model of interaction with an intestinal cell line. In general, 1-kestose was the compound showing the largest effects. The modulation with prebiotics led to significant increases in the Bacteroides group and Faecalibacterium in obese subjects, whereas in normal-weight individuals, substantial rises in Bifidobacterium and Faecalibacterium were appreciated. Notably, the results obtained showed differences in the responses among the tested compounds but also among the studied human populations, indicating the need for developing population-specific products.

Keywords: HT29; RTCA; SCFA; bifidobacterial-ITS; functionality; gas production; in vitro model; microbiota; obesity; prebiotics.

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Conflict of interest statement

Takumi Tochio and Katsuaki Hirano are employees at the company β- Food Sciences.

Figures

Figure 1
Figure 1
Microbial composition (relative abundance %) evaluated by 16S rRNA gene profiling at family levels in basal conditions (time 0: T0) and after 24 h of incubation in fecal cultures with several carbon sources and without an external carbon source added (Control) in morbid obesity (MOB) and normal-weight (NW) groups.
Figure 2
Figure 2
Absolute levels (Log CFU/mL) of fecal microbial groups determined by qPCR after fecal cultures of (A) MOB and (B) NW subjects. For each microbial group, the box and whiskers plot represent median, interquartile range and minimum and maximum values obtained in each human group (NW or MOB). Different letters above the boxes indicate significant differences (p-value < 0.05) among carbon sources for the microbial groups considered.
Figure 3
Figure 3
Increments in ascending order, with respect to time 0, in the concentration (mM) of the major short-chain fatty acids (acetic, propionic and butyric) after 24 h of incubation with different carbon sources in fecal cultures from MOB (A) and NW (B) groups. Differences are shown for each short-chain fatty acid (SCFA); columns that do not share the same letter are significantly different (p < 0.05).
Figure 4
Figure 4
Real-time monitoring the interaction between (A) fecal supernatants and (B) isolated microbiota obtained before and after incubation with prebiotics and HT29 intestinal epithelial cells. Values (media ± SD) correspond to the Area Under the Curve (AUC) resulting from monitoring the cell index (CI) during 10 h. Significant differences (p-value < 0.05) represent the comparison of results before and after prebiotics addition in each condition.
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