scholarly journals Pathogen Challenge and Dietary Shift Alter Microbiota Composition and Activity in a Mucin-Associated in vitro Model of the Piglet Colon (MPigut-IVM) Simulating Weaning Transition

2021 ◽  
Vol 12 ◽  
Author(s):  
Raphaële Gresse ◽  
Frédérique Chaucheyras-Durand ◽  
Juan J. Garrido ◽  
Sylvain Denis ◽  
Angeles Jiménez-Marín ◽  
...  
Keyword(s):  
Intestinal Inflammation ◽  
Animal Experiments ◽  
Short Chain Fatty Acids ◽  
Stress Factors ◽  
Intestinal Cell ◽  
Gut Health ◽  
Dietary Shift ◽  
Pathogen Challenge ◽  
Weaning Piglets

Enterotoxigenic Escherichia coli (ETEC) is the principal pathogen responsible for post-weaning diarrhea in newly weaned piglets. Expansion of ETEC at weaning is thought to be the consequence of various stress factors such as transient anorexia, dietary change or increase in intestinal inflammation and permeability, but the exact mechanisms remain to be elucidated. As the use of animal experiments raise more and more ethical concerns, we used a recently developed in vitro model of piglet colonic microbiome and mucobiome, the MPigut-IVM, to evaluate the effects of a simulated weaning transition and pathogen challenge at weaning. Our data suggested that the tested factors impacted the composition and functionality of the MPigut-IVM microbiota. The simulation of weaning transition led to an increase in relative abundance of the Prevotellaceae family which was further promoted by the presence of the ETEC strain. In contrast, several beneficial families such as Bacteroidiaceae or Ruminococcaceae and gut health related short chain fatty acids like butyrate or acetate were reduced upon simulated weaning. Moreover, the incubation of MPigut-IVM filtrated effluents with porcine intestinal cell cultures showed that ETEC challenge in the in vitro model led to an increased expression of pro-inflammatory genes by the porcine cells. This study provides insights about the etiology of a dysbiotic microbiota in post-weaning piglets.

scholarly journals Gluten-induced RNA methylation changes regulate intestinal inflammation via allele-specific XPO1 translation in epithelial cells

Gut ◽  
2021 ◽  
pp. gutjnl-2020-322566
Author(s):  
Ane Olazagoitia-Garmendia ◽  
Linda Zhang ◽  
Paula Mera ◽  
Julie K Godbout ◽  
Maialen Sebastian-DelaCruz ◽  
...  
Keyword(s):  
Intestinal Inflammation ◽  
Autoimmune Disorder ◽  
Intestinal Cell ◽  
In Vivo Models ◽  
Knock Out ◽  
New Therapeutic Approaches ◽  
Intestinal Cell Line ◽  
Environment Characteristic

ObjectivesCoeliac disease (CD) is a complex autoimmune disorder that develops in genetically susceptible individuals. Dietary gluten triggers an immune response for which the only available treatment so far is a strict, lifelong gluten free diet. Human leucocyte antigen (HLA) genes and several non-HLA regions have been associated with the genetic susceptibility to CD, but their role in the pathogenesis of the disease is still essentially unknown, making it complicated to develop much needed non-dietary treatments. Here, we describe the functional involvement of a CD-associated single-nucleotide polymorphism (SNP) located in the 5’UTR of XPO1 in the inflammatory environment characteristic of the coeliac intestinal epithelium.DesignThe function of the CD-associated SNP was investigated using an intestinal cell line heterozygous for the SNP, N6-methyladenosine (m6A)-related knock-out and HLA-DQ2 mice, and human samples from patients with CD.ResultsIndividuals harbouring the risk allele had higher m6A methylation in the 5’UTR of XPO1 RNA, rendering greater XPO1 protein amounts that led to downstream nuclear factor kappa B (NFkB) activity and subsequent inflammation. Furthermore, gluten exposure increased overall m6A methylation in humans as well as in in vitro and in vivo models.ConclusionWe identify a novel m6A-XPO1-NFkB pathway that is activated in CD patients. The findings will prompt the development of new therapeutic approaches directed at m6A proteins and XPO1, a target under evaluation for the treatment of intestinal disorders.

scholarly journals In Vitro Fecal Fermentation Patterns of Arabinoxylan from Rice Bran on Gut Microbiota in Normal Weight and Overweight/Obese Subjects

2020 ◽  
Vol 4 (Supplement_2) ◽  
pp. 1560-1560
Author(s):  
Inah Gu ◽  
Wing Shun Lam ◽  
Daya Marasini ◽  
Cindi Brownmiller ◽  
Brett Savary ◽  
...  
Keyword(s):  
Gut Microbiota ◽  
Rice Bran ◽  
Block Design ◽  
16S Rrna Gene Sequencing ◽  
Short Chain Fatty Acids ◽  
Normal Weight ◽  
Rrna Gene ◽  
Gut Health ◽  
The Impact

Abstract Objectives Arabinoxylan is a non-starch polysaccharide and rich in wheat, rice and many other cereal grains. Diets high in fiber help promoting gut health in obesity. The objective of this study was to investigate the impact of arabinoxylan from rice bran on the gut microbiota and short chain fatty acids (SCFA) in normal weight (NW) and overweight/obese (OO) subjects through in vitro fecal fermentation. Methods Arabinoxylan was extracted from rice bran fiber. For in vitro fecal fermentation, each fecal sample from NW (n = 6, 3 males and 3 females) and OO (n = 7, 3 males and 4 females) was diluted into anaerobic medium with three treatments: control (no substrates), fructooligosaccharides (FOS, a well-known prebiotic), and arabinoxylan. Samples were incubated at 37˚C and aliquots were taken at 0, 4, 8, 12 and 24 h. SCFA content from samples at all timepoints was analyzed using HPLC. Samples at 0 and 24 h were used for gut microbiota analysis through 16S rRNA gene sequencing. Statistical analyses were performed for the randomized complete block design, where the weight classes are confounded with blocks (subjects). Friedman test was used to determine the difference at 5% level of significance. Results As a result, arabinoxylan treatment significantly increased total SCFA concentration in both NW and OO subjects than control (P < 0.05), comparable to FOS treatment. Between weight classes under arabinoxylan treatment, OO group showed a significantly higher total SCFA content than NW group (P < 0.05). Arabinoxylan changed gut microbial population at the genus level, stimulating Bifidobacterium, Collinsella and Blautia and decreasing Clostridium XIVa and b, Dorea and Oscillibacter (P < 0.05). In addition, different microbiome population was shown in weight classes with three treatments, showing higher Bacteroides in NW and higher Prevotella in OO. Conclusions These results showed that arabinoxylan from rice bran modified gut microbiota in both weight classes, increasing total SCFA content. This study suggests that arabinoxylan from rice bran may have a potential impact on microbial gut health in obesity with prebiotic activities. Funding Sources University of Arkansas.

scholarly journals Assessing the Effects of Ginger Extract on Polyphenol Profiles and the Subsequent Impact on the Fecal Microbiota by Simulating Digestion and Fermentation In Vitro

Nutrients ◽  
2020 ◽  
Vol 12 (10) ◽  
pp. 3194
Author(s):  
Jing Wang ◽  
Yong Chen ◽  
Xiaosong Hu ◽  
Fengqin Feng ◽  
Luyun Cai ◽  
...  
Keyword(s):  
Gut Microbiota ◽  
Ph Value ◽  
Illumina Miseq ◽  
Short Chain Fatty Acids ◽  
Fecal Microbiota ◽  
Rrna Gene ◽  
Gut Health ◽  
Ginger Extract ◽  
Mixed Culture Fermentation

The beneficial effects of ginger polyphenols have been extensively reported. However, their metabolic characteristics and health effects on gut microbiota are poor understood. The purpose of this study was to investigate the digestion stability of ginger polyphenols and their prebiotic effects on gut microbiota by simulating digestion and fermentation in vitro. Following simulated digestion in vitro, 85% of the polyphenols were still detectable, and the main polyphenol constituents identified in ginger extract are 6-, 8-, and 10-gingerols and 6-shogaol in the digestive fluids. After batch fermentation, the changes in microbial populations were measured by 16S rRNA gene Illumina MiSeq sequencing. In mixed-culture fermentation with fecal inoculate, digested ginger extract (GE) significantly modulated the fecal microbiota structure and promoted the growth of some beneficial bacterial populations, such as Bifidobacterium and Enterococcus. Furthermore, incubation with GE could elevate the levels of short-chain fatty acids (SCFAs) accompanied by a decrease in the pH value. Additionally, the quantitative PCR results showed that 6-gingerol (6G), as the main polyphenol in GE, increased the abundance of Bifidobacterium significantly. Therefore, 6G is expected to be a potential prebiotic that improves human health by promoting gut health.

scholarly journals Impact of Grain Sorghum Polyphenols on Microbiota of Normal Weight and Overweight/Obese Subjects during In Vitro Fecal Fermentation

Nutrients ◽  
2019 ◽  
Vol 11 (2) ◽  
pp. 217 ◽  
Author(s):  
Danielle Ashley ◽  
Daya Marasini ◽  
Cindi Brownmiller ◽  
Jung Lee ◽  
Franck Carbonero ◽  
...  
Keyword(s):  
Gut Microbiota ◽  
Batch Fermentation ◽  
Short Chain Fatty Acids ◽  
Negative Control ◽  
Normal Weight ◽  
Weight Class ◽  
Gut Health ◽  
Sorghum Bran ◽  
Before And After

The human gut microbiota is considered as a crucial mediator between diet and gut homeostasis and body weight. The unique polyphenolic profile of sorghum bran may promote gastrointestinal health by modulating the microbiota. This study evaluated gut microbiota and modulation of short-chain fatty acids (SCFA) by sorghum bran polyphenols in in vitro batch fermentation derived from normal weight (NW, n = 11) and overweight/obese (OO, n = 11) subjects’ fecal samples. Six separate treatments were applied on each batch fermentation: negative control (NC), fructooligosaccharides (FOS), black sorghum bran extract (BSE), sumac sorghum bran extract (SSE), FOS + BSE, or FOS + SSE; and samples were collected before and after 24 h. No significant differences in total and individual SCFA production were observed between NW and OO subjects. Differential responses to treatment according to weight class were observed in both phyla and genera. Sorghum bran polyphenols worked with FOS to enhance Bifidobacterium and Lactobacillus, and independently stimulated Roseburia and Prevotella (p < 0.05). Our results indicate that sorghum bran polyphenols have differential effects on gut health and may positively impact gut ecology, with responses varying depending on weight class.

scholarly journals Xylo-oligosaccharides from sugarcane show prebiotic potential in a dynamic computer-controlled in vitro model of the adult human large intestine

2020 ◽  
Vol 11 (2) ◽  
pp. 191-200 ◽  
Author(s):  
K. Venema ◽  
J. Verhoeven ◽  
S. Verbruggen ◽  
D. Keller
Keyword(s):  
In Vitro Model ◽  
Short Chain Fatty Acids ◽  
Rrna Gene ◽  
Microbiota Composition ◽  
Gut Health ◽  
Unidentified Species ◽  
Human Adults ◽  
Computer Controlled ◽  
Vitro Model

The aim of the study was to investigate the prebiotic potential of xylo-oligosaccharides (XOS) from sugarcane in a validated, dynamic, computer-controlled in vitro model of the colon (TIM-2) simulating human adults. In two sets of experiments, each with a different microbiota, 3 different doses of XOS were tested at 1.0 g/day, 1.5 g/day and 3.0 g/day. The in vitro model was run for 72 h, and at the start and subsequently every 24 h samples were taken and analysed for short-chain fatty acids (SCFA) and gut microbiota composition. SCFA were analysed using ion chromatography, whereas microbiota composition was analysed using sequencing of the V3-V4 region of the 16S rRNA gene. XOS showed a similar SCFA production per gram of substrate as the control medium, including butyrate, which is considered to be important for gut health. In both sets of experiments XOS showed a consistent dose-dependent increase in abundance over time of the genus Bifidobacterium, and within that of the species B. adolescentis and an unidentified species (labelled ‘sp.1’). The results show the potential prebiotic effect of XOS from sugarcane, by its capacity to generate butyrate and increase the health-beneficial bifidobacteria.

scholarly journals Characterization of Kazachstania slooffiae, a Proposed Commensal in the Porcine Gut

2021 ◽  
Vol 7 (2) ◽  
pp. 146
Author(s):  
Katie Lynn Summers ◽  
Juli Foster Frey ◽  
Ann M. Arfken
Keyword(s):  
Genetic Relatedness ◽  
Growth Parameters ◽  
Fungal Growth ◽  
Short Chain Fatty Acids ◽  
Growth Conditions ◽  
Antifungal Drugs ◽  
Positive Interactions ◽  
Gut Health

Kazachstania slooffiae is a fungus commonly isolated from the gastrointestinal tract and feces of post-weaning pigs. Studies have implicated its ability to positively alter piglet gut health through potential symbioses with beneficial bacteria, including Lactobacillus and Prevotella, in providing amino acids as an energy source for microbial and piglet growth, and it has been found to be positively correlated with short-chain fatty acids in the piglet gut. However, basic mycological information remains limited, hampering in vitro studies. In this study, we characterized the growth parameters, biofilm formation ability, susceptibility to antimicrobials, and genetic relatedness of K. slooffiae to other fungal isolates. Optimal fungal growth conditions were determined, no antifungal resistance was found against multiple classes of antifungal drugs (azoles, echinocandins, polyenes, or pyrimidine analogues), and dimorphic growth was observed. K. slooffiae produced biofilms that became more complex in the presence of Lactobacillus acidophilus supernatant, suggesting positive interactions with this bacterium in the gut, while Enterococcus faecalis supernatant decreased density, suggesting an antagonistic interaction. This study characterizes the in vitro growth conditions that are optimal for further studies of K. slooffiae, which is an important step in defining the role and interactions of K. slooffiae in the porcine gut environment.

scholarly journals PSIII-18 Assessment of Salmonella Dublin infection of intestinal porcine epithelial cells (IPEC) in response to Zinc Oxide and a Yeast Mannan Rich Fraction

2020 ◽  
Vol 98 (Supplement_4) ◽  
pp. 365-366
Author(s):  
Niall Browne ◽  
Seána Jordan ◽  
Karina Horgan
Keyword(s):  
Gene Expression ◽  
Zinc Oxide ◽  
Tight Junction ◽  
Control Cell ◽  
Bacterial Attachment ◽  
Intestinal Cell ◽  
Gut Health ◽  
Suitable Alternative ◽  
Salmonella Dublin

Abstract Salmonella species are associated with post-weaning diarrhea, which results in poor weight gain, potential death, and economic cost. A yeast mannan rich fraction (MRF) was assessed alongside the industry standard treatment Zinc Oxide (ZO) in vitro to determine its impact on Salmonella Dublin infection of a pig intestinal cell line (IPEC). IPEC cells were exposed to MRF or ZO in the presence of S. Dublin (1x108/mL). IPEC cell RNA was isolated and cDNA synthesized. Gene expression for IL-1β, TNFα, IL-8 and cellular tight Junction genes Occludin, Claudin3 and Tight junction protein1 (TJP1) were assessed by qPCR. S. Dublin adhesion to IPEC cells (500:1) assessed in the presence or absence of either ZO or MRF for 1 hour at 37°C. IPEC cells with attached S. Dublin were lysed, diluted, plated and incubated overnight and enumeration. Three biological replicates were performed for all experiments and statistical analysis determined by One-way ANOVA. Proinflammatory gene TNFα was significantly reduced (P ≤ 0.001) following S. Dublin infection and treatment with MRF compared with ZO. IL-1β demonstrated no change between treatments although IL-8 gene expression was significantly reduced in both ZO (P≤0.01) and MRF (P ≤ 0.05) treated cells over the control. Significantly higher expression of Occludin (P ≤ 0.01), Claudin3 (P ≤ 0.001) and TJP1 (P ≤ 0.05) was observed in IPEC cells exposed to S. Dublin in the presence of MRF compared to ZO. Adhesion of S. Dublin to IPEC cells was significantly reduced in response to MRF addition compared to ZO treated cells (P ≤ 0.001) and the control cells (P ≤ 0.05). ZO treated cell demonstrated no improvement over the control cell levels of bacterial attachment. Both on a physical and molecular level bacterial infection of intestinal cells was more significantly impaired by MRF addition. With the ban on ZO, yeast MRF may prove to be a suitable alternative to support gut health in piglets.

scholarly journals Prebiotic Dietary Fiber and Gut Health: Comparing the In Vitro Fermentations of Beta-Glucan, Inulin and Xylooligosaccharide.

2017 ◽  
Author(s):  
Justin L. Caelson ◽  
Jennifer M. Erickson ◽  
Julie M. Hess ◽  
Trevor J. Gould ◽  
Joanne L. Slavin
Keyword(s):  
Dietary Fiber ◽  
Short Chain Fatty Acids ◽  
Fecal Microbiota ◽  
Gas Production ◽  
Gut Health ◽  
Beta Glucan ◽  
In Vitro Fermentation ◽  
Gastrointestinal Health ◽  
Oat Bran

Prebiotic dietary fiber supplements are commonly consumed to help meet fiber recommendations and improve gastrointestinal health by stimulating beneficial bacteria and the production of short-chain fatty acids (SCFAs), molecules beneficial to host health. The objective of this research project was to compare potential prebiotic effects and fermentability of five commonly consumed fibers using an in vitro fermentation system measuring changes in fecal microbiota, total gas production and formation of common SCFAs. Fecal donations were collected from three healthy volunteers. Materials analyzed included: pure beta-glucan, Oatwell (commercially available oat-bran containing 22% oat &beta;-glucan), xylooligosaccharides (XOS), WholeFiber (dried chicory root containing inulin, pectin, and hemi/celluloses), and pure inulin. Oatwell had the highest production of propionate at 12 h (4.76 &mu;mol/mL) compared to inulin, WholeFiber and XOS samples (p&lt;0.03). Oatwell&rsquo;s effect was similar to those of the pure beta-glucan samples, both samples promoted the highest mean propionate production at 24 h. XOS resulted in a significant increase in the genus Bifidobacterium after 24 h of fermentation (0 h: 0.67 OTUs; 24 h: 5.22 OTUs; p = 0.038). Inulin and WholeFiber increased the beneficial genus Collinsella, consistent with findings in clinical studies. All analyzed compounds were fermentable and promoted the formation of beneficial SCFAs.

scholarly journals Development of Anti-inflammatory Probiotic Limosilactobacillus reuteri EFEL6901 as Kimchi Starter: in vitro and In vivo Evidence

2021 ◽  
Vol 12 ◽  
Author(s):  
Hee Seo ◽  
Hyunbin Seong ◽  
Ga Yun Kim ◽  
Yu Mi Jo ◽  
Seong Won Cheon ◽  
...  
Keyword(s):  
Activity Index ◽  
Body Weight Loss ◽  
Disease Activity Index ◽  
Short Chain Fatty Acids ◽  
Fermented Foods ◽  
Gut Health ◽  
Gut Barrier Function ◽  
Anti Inflammatory

The use of probiotic starters can improve the sensory and health-promoting properties of fermented foods. In this study, we developed an anti-inflammatory probiotic starter, Limosilactobacillus reuteri EFEL6901, for use in kimchi fermentation. The EFEL6901 strain was safe for use in foods and was stable under human gastrointestinal conditions. In in vitro experiments, EFEL6901 cells adhered well to colonic epithelial cells and decreased nitric oxide production in lipopolysaccharide-induced macrophages. In in vivo experiments, oral administration of EFEL6901 to DSS-induced colitis mice models significantly alleviated the observed colitis symptoms, prevented body weight loss, lowered the disease activity index score, and prevented colon length shortening. Analysis of these results indicated that EFEL6901 played a probiotic role by preventing the overproduction of pro-inflammatory cytokines, improving gut barrier function, and up-regulating the concentrations of short-chain fatty acids. In addition, EFEL6901 made a fast growth in a simulated kimchi juice and it synthesized similar amounts of metabolites in nabak-kimchi comparable to a commercial kimchi. This study demonstrates that EFEL6901 can be used as a suitable kimchi starter to promote gut health and product quality.

scholarly journals In Vitro Digestion and Fermentation by Human Fecal Microbiota of Polysaccharides from Flaxseed

Molecules ◽  
2020 ◽  
Vol 25 (19) ◽  
pp. 4354
Author(s):  
Xin Zhou ◽  
Zhao Zhang ◽  
Fenghong Huang ◽  
Chen Yang ◽  
Qingde Huang
Keyword(s):  
Gut Microbiota ◽  
Reducing Sugars ◽  
In Vitro Digestion ◽  
Short Chain Fatty Acids ◽  
Fecal Microbiota ◽  
Gut Health ◽  
Human Gut ◽  
In Vitro Fermentation ◽  
Composition And Structure

The digestion of flaxseed polysaccharides (FSP) in simulated saliva, gastric and small intestine conditions was assessed, as well as in vitro fermentation of FSP by human gut microbiota. FSP was not degraded in the simulated digestive systems (there was no change in molecular weight or content of reducing sugars), indicating that ingested FSP would reach the large intestine intact. Changes in carbohydrate content, reducing sugars and culture pH suggested that FSP could be broken down and used by gut microbiota. FSP modulated the composition and structure of the gut microbiota by altering the Firmicutes/Bacteroidetes ratio and increasing the relative abundances of Prevotella, Phascolarctobacterium, Clostridium and Megamonas, which can degrade polysaccharides. Meanwhile, FSP fermentation increased the concentration of short-chain fatty acids, especially propionic and butyric acids. Our results indicate that FSP might be developed as a functional food that benefits gut health.

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