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

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.

Does intestinal dysbiosis contribute to an aberrant inflammatory response to severe acute respiratory syndrome coronavirus 2 in frail patients?

“…Zhang Q et al have demonstrated that inulin decreases Desulfovibrio abundance, reducing LPS and IL-6 levels in mice models [ 83 , 84 ] . This inulin action is replicated in human clinical trials [ 85 , 86 ] .…”

In vitro Selection of Probiotics for Microbiota Modulation in Normal-Weight and Severely Obese Individuals: Focus on Gas Production and Interaction With Intestinal Epithelial Cells

“…In brief, seven independent pH-free batch fermentations were performed for each human donor. We used as a carbon source 0.3% (w/v) of the fructooligosaccharide 1-kestose (β-Food Science Co. Ltd., Japan) which in previous experiments demonstrated to be more fermentable than other fructooligosaccharides not just by bifidobacteria and other intestinal anaerobes but also by lactobacilli ( Nogacka et al, 2020b ). Bacterial strains were added at a final concentration of 1 × 10 8 CFU/mL to fecal cultures in bottles.…”

“…Different models of GIM-host interaction, such as co-cultivation of fecal samples or isolated GIM from different population groups or added with pro/prebiotics, with epithelial and/or immune cells, have been used ( Arboleya et al, 2015 ; Nogacka et al, 2018 , 2020a ; Richards et al, 2019 ). Interestingly, several of these studies have demonstrated differences in the response induced by microbiotas from different population groups ( Nogacka et al, 2018 , 2020a ) and between isolated GIM or among GIM added with probiotics or prebiotics ( Arboleya et al, 2015 ; Nogacka et al, 2020b ). These suggest that the inter-population differences on the gut microbiota may partly explain the high variability observed in the response to probiotics and the low correlation between in vivo and in vitro data.…”

Comparison of Different Dietary Indices as Predictors of Inflammation, Oxidative Stress and Intestinal Microbiota in Middle-Aged and Elderly Subjects

“…After deposition, samples were immediately frozen at −20 °C and transported to the laboratory. For analyses, samples were thawed at room temperature (24 ± 2 °C), weighed, diluted 1/10 in sterile PBS and homogenized using a LabBlender 400 Stomacher (Seward Medical, London, UK) for 4 min; the DNA was extracted using the QIAamp DNA stool mini kit (Qiagen, Hilden, Germany) following previously described procedures [ 41 ]. 7500 Fast Real-Time PCR System (Applied Biosystems, Foster City, CA, USA) and SYBR Green PCR Master Mix (Applied Biosystems) were used to achieve the quantification of bacterial populations, including the major bacterial groups present in the human gut ( Supplementary Table S2 ).…”

“…7500 Fast Real-Time PCR System (Applied Biosystems, Foster City, CA, USA) and SYBR Green PCR Master Mix (Applied Biosystems) were used to achieve the quantification of bacterial populations, including the major bacterial groups present in the human gut ( Supplementary Table S2 ). Procedure instructions were published elsewhere [ 41 ]. Fecal DNA extracts were analyzed and the mean quantity per gram of fecal wet weight was calculated for each bacterial group.…”