Total Short-chain Fatty Acid Production Research Articles

Flavones interact with fiber to affect fecal bacterial communities in vitro

This study investigated the effects of the sugarcane flavones diosmin, diosmetin, luteolin, and tricin, and their interactions with sugarcane fiber on the modulation of gut microbiota using in vitro batch fermentation. The alteration of fecal fermentation bacterial profile was analyzed using 16S rRNA sequencing data, while the bioavailability of fiber was indicated by short-chain fatty acid (SCFA) production and metabolism of polyphenols was measured directly by phenolic metabolites. Application of diosmin, diosmetin, luteolin, and tricin without fiber had no significant effect on the overall microbiota profile after 24 h of fermentation. When fiber alone was added, total SCFA production increased, specifically that of propionic and valeric acids. However, when flavones were combined with fiber, synergistic effects on the modulation of relative abundances of different bacterial taxa was noted. In particular, the proportion of Prevotella spp. was significantly increased by the combinations of diosmin, luteolin, and tricin with fiber.

In Vitro Colonic Fermentation Profiles and Microbial Responses of Cellulose Derivatives with Different Colloidal States.

Although cellulose derivatives are widely applied in the food industry, the effects of their structural properties on colonic health is unknown. Here, four types of cellulose derivatives, including microcrystalline cellulose (MCC), TEMPO-oxidized nanofibrillated cellulose (TOCNF), TEMPO-oxidized nanocrystalline cellulose (TOCNC), and carboxymethyl cellulose (CMC) were selected to investigate their in vitro fermentation profiles. TOCNF exhibited the highest production of total short-chain fatty acids (SCFAs), followed by TOCNC. The results suggested that reduced particle size and increased aspect ratio improved the fermentability of insoluble cellulose derivatives. MCC and CMC were barely fermented with similar total SCFAs production as the blank. 16S rRNA sequencing revealed that the fermentation of cellulose derivatives resulted in divergent microbial community structures. Moreover, Bacteroides cellulosilyticus showed high specificity to utilize TOCNF and TOCNC. The findings demonstrated that the colloidal states of cellulose derivatives, such as size and solubility, were important factors governing microbial community composition and metabolites.

Assessment of the bioaccessibility of phenolics from Australian grown lettuces by in vitro simulated gastrointestinal digestion and colonic fermentation

Lettuce varieties are rich in phytochemicals with strong antioxidant properties, mainly due to the presence of many phenolic compounds. The health effects of these phenolic compounds are dependent on their bioaccessibility, bioavailability and bioactivities. This study aimed to evaluate the differences in the content and bioaccessibility of phenolic compounds in green lettuce (iceberg, baby cos, and green little gem) and red lettuce (red oak leaf, lollo rosso, and red little gem), their antioxidant properties and short chain fatty acids (SCFAs) production after in vitro gastrointestinal digestion and colonic fermentation. Lollo rosso showed the highest content of phenolic compounds (1.23 mg GAE/g) and flavonoids (0.36 mg QE/g), as well as the antioxidant capacities after gastrointestinal digestion. After colonic fermentation, only baby cos and red little gem showed significantly higher TPC (5.13 mg GAE/g) and TFC (1.95 mg GAE/g), respectively. Most phenolic compounds in lettuce were mostly bioaccessible after gastric and small intestinal digestion, except for the kaempferol which relied more on the colonic fermentation. However, high percentage of kaempferol still could be found in the colonic fermented residual, probably due to the presence of dietary fiber and the interactions with other components. All lettuce varieties achieved the peak production of SCFAs after 16 h, except iceberg which displayed the highest total SCFAs production just after 8 h fermentation. It could be inferred that the iceberg (green lettuce) could be more beneficial to gut health while lollo rosso (red lettuce) could contribute more antioxidant properties after consumption.

Starchy and fibrous feedstuffs differ in their in vitro digestibility and fermentation characteristics and differently modulate gut microbiota of swine

BackgroundAlternative feedstuffs may contribute to reducing feed costs of pig production. But these feedstuffs are typically rich in fiber and resistant starch (RS). Dietary fibers and RS are fermented in the gastrointestinal tract (GIT) and modulate the microbial community. Certain microbes in the GIT can promote host health, depending on the type of fermentation substrates available. In this study, six alternative feedstuffs (three starchy: Okinawan sweet potato, OSP; yam, and taro, and three fibrous: wheat millrun, WMR; barley brewers grain, BBG; and macadamia nut cake, MNC) were evaluated for their in vitro digestibility and fermentation characteristics and their effects on pig’s hindgut microbial profile. After 2 steps of enzymatic digestion assay, residues were fermented using fresh pig feces as microbial inoculum, and gas production was recorded periodically for 72 h and modeled for fermentation kinetics. After fermentation, the residual liquid phase was analyzed for short-chain fatty acid (SCFA), and the solid phase was used to determine the nutrient’s digestibility and microbial community.ResultsIn vitro ileal digestibility of dry matter and gross energy was higher in starchy than fibrous feedstuffs. Total gas and SCFA production were significantly higher (P < 0.001) in starchy feedstuffs than fibrous feedstuffs. Both acetate and propionate production was significantly higher (P < 0.001) in all starchy feedstuffs than BBG and MNC; WMR was in between. Overall alpha diversity was not significantly different within and between starchy and fibrous feedstuffs. Beta diversity (measured using bray Curtis dissimilarity distance) of starchy feedstuffs was significantly different (P < 0.005) than fibrous feedstuffs.ConclusionStarchy feedstuffs acted as a substrate to similar types of microbes, whereas fibrous feedstuffs resulted in a more diverse microbial population. Such alternative feedstuffs may exert comparable beneficial effects, thus may be included in swine diets to improve gut health.

Hermetia illucens fat affects the gastrointestinal tract selected microbial populations, their activity, and the immune status of broiler chickens

Abstract The present study investigated the effect of Hermetia illucens larvae (BSFL) fat, derived using supercritical CO2 extraction and added to broiler chickens’ diets as a partial (50%) or total replacement for commonly used soybean oil, on the gastrointestinal tract (GIT) microbial population, its activity, and selected physiological and immune traits. A total of 576 one-day-old female Ross 308 chicks were randomly assigned to 3 dietary treatments with 16 replicates each. The following treatments were applied: SO – 100% soybean oil, BSFL50 – a mixture of BSFL and soybean oils in a 50:50 ratio, and BSFL100 – 100% BSFL fat. Digesta samples from the crop, jejunum and caeca were collected for further analyses, i.e., pH measurements, fluorescent in situ hybridization, and short-chain fatty acid (SCFA) concentrations. Additionally, the selected plasma biochemical parameters and immunological traits were assessed. In general, the implementation of BSFL fat in broilers’ diets resulted in increased proliferation of potentially pathogenic bacterial populations in the crop, such as Enterobacteriaceae, Bacteroides–Prevotella cluster, and Clostridium perfringens. Furthermore, BSFL100 enhanced microbial activity via total SCFA production and lowered the pH in this segment. However, no detrimental effects were observed in terms of other GIT segments, i.e., the jejunal and cecal microecosystems. The strongest impact on reduction of select components of the microbial population in the cecum was observed with the BSFL50 treatment for potentially pathogenic bacteria such as Enterobacteriaceae, Bacteroides–Prevotella cluster, while commensal populations were also limited, i.e., Bacillus spp., C. leptum subgroup, and C. coccoides–Eubacterium rectale cluster. Additionally, BSFL100 reduced the cholesterol concentration in the blood, while both experimental treatments decreased the ALT level. In conclusion, due to the insufficient release of lauric acid from the BSFL fat in the crop, an adverse shift in the microbiota can be noted. However, a positive suppressive effect on the select components of the cecal microbiota, as well as improvement of liver health suggests implying the BSFL fat in broiler nutrition.

Bioaccessibility and bioactivities of phenolic compounds from roasted coffee beans during in vitro digestion and colonic fermentation

Bioaccessibility and bioactivity of phenolic compounds in coffee beans relate to roasting and digestion process. This study aimed to estimate phenolic content, antioxidant potential, bioaccessibility, and changes in short chain fatty acids (SCFAs) production during in vitro digestion and colonic fermentation of commercial roasted (light, medium and dark) coffee beans. There was no significant difference found among all three different roasting levels. TPC and DPPH were enhanced 15 mg GAE/g and 60 mg TE/g during gastrointestinal digestion, respectively. For colonic fermentation, the highest TPC and FRAP of all coffee beans was found at 2 and 4 h, respectively. The gastric bioaccessibility of most of the phenolic compounds were relatively higher due to thermal phenolic degradation. Total SCFAs production was only up to 0.02 mM because of thermal polysaccharide decomposition. Light roasted beans exhibited relatively higher phenolic bioaccessibility, antioxidant activities and SCFAs production, which would be more beneficial to gut health.

In vitro fecal fermentation characteristics of bamboo insoluble dietary fiber and its impacts on human gut microbiota

Bamboo contains abundant hemicellulose, cellulose, and lignin, which are a high-quality insoluble dietary fiber (IDF) raw material. To investigate IDF- induced changes in the production of short-chain fatty acids (SCFAs) and the composition of human gut microbiota, IDF was extracted by alkaline hydrogen peroxide (named BIDF), complex enzymatic hydrolysis method (named OIDF) from bamboo, and commercial bamboo fiber BF90. The in vitro fecal fermentation characteristics of BIDF, OIDF, BF90 and its impacts on human gut microbiota were studied for the first time. Results showed that BIDF, OIDF, and BF90 could promote the production of total SCFAs after 24 h fermentation. Additionally, BIDF could alter the composition and microbial diversity of gut microbiota, especially increase the relative abundance of Bacteroides and decrease the ratio of Firmicutes to Bacteroidetes (F/B) value. These results revealed that the IDF from bamboo could be partially utilized by specific bacteria in human intestines and provide a reference for the study of the effects of IDF fermentation on SCFAs production and microbial composition.

Sugarcane polyphenol and fiber to affect production of short-chain fatty acids and microbiota composition using in vitro digestion and pig faecal fermentation model

This study aimed to examine the effects of sugarcane polyphenol and fiber (Phytolin + Fiber) on gut microbiota, short-chain fatty acids (SCFAs) production and phenolic metabolites production using in vitro digestion and fermentation model. Microbial profiling by 16S rRNA sequencing was used to analyze the pig faecal microbiota profile. SCFAs were identified and quantified by GC-FID, and phenolic metabolites were characterized by LC-ESI-QTOF-MS/MS. The results showed that Phytolin + Fiber exert synergistic effects on the pig gut microbiota by increasing the relative abundances of Lactobacillus and Catenibacterium, and decreasing the relative abundances of Mogibacterium, Dialister, and Escherichia-Shigella. Phytolin + Fiber also significantly increased the total SCFAs production, particularly the propionic and butyric acids. Production of phenolic metabolites related to major polyphenols in Phytolin were tentatively identified. These results suggest that Phytolin + Fiber could be beneficial to human colon health given the similarities between pig and human intestine in terms of physiology and microbiome.

Effects of in ovo delivered xylo- and mannan- oligosaccharides on growth performance, intestinal immunity, cecal short-chain fatty acids, and cecal microbiota of broilers

BackgroundThis study investigated a novel in ovo feeding strategy to determine the prebiotic effects of xylo- and mannan- oligosaccharides (XOS and MOS) differing in the degree of polymerization. A total of 192 fertilized eggs were divided into 6 treatment groups: i) normal saline control (NSC), ii) xylotriose (XOS3), iii) xylotetraose (XOS4), iv) mannotriose (MOS3), v) mannotetraose (MOS4), and vi) no injection control (NIC), each containing 4 replicate trays with 8 eggs per replicate. On d 17 of incubation, 3 mg of oligosaccharides (except for controls) dissolved in 0.5 mL of 0.85% normal saline were injected into the amnion of Cobb 500 broilers eggs. After hatch, the chicks were raised for 28 d under standard husbandry practices and were fed a commercial broilers diet ad libitum, and samples were collected periodically.ResultsThe hatchability, growth performance, and relative weights of breast, drumstick, liver, and proventriculus were not different among the treatments (P > 0.05). The XOS3 injection increased the total short-chain fatty acid production at d 28 compared with both control groups (P < 0.05). The villus height to crypt depth ratio was significantly higher in the XOS4 group than both controls on the hatch day (P < 0.01) but were not different among any treatments on d 7 and 28 (P > 0.05). On the hatch day, the expression level of the CD3 gene (a T cell marker) was increased by XOS3, while the IL-10 gene (a marker of anti-inflammatory cytokine) was reduced by MOS4 (P < 0.05) compared with both controls. Compared with both controls, XOS3 exhibited a trend of reduction for IL-10 (P = 0.074). No cytokines or lymphocyte markers were affected by the treatments on d 7 (P > 0.05), except XOS4 increased IL-4 compared with NSC (P < 0.05). The broilers in the MOS4 group had higher operational taxonomic units (OTUs) and had more differentially abundant taxa, including order Lactobacillales and family Leuconostocaceae (P < 0.05) than both controls on d 28. The predictive functional profiling indicated that the linoleic acid metabolism pathway was enriched in the cecal microbiota of the XOS3 group compared with both controls (P < 0.05).ConclusionThe effects of these XOS and MOS on ileal mucosa and immunity are transient, but the effects on fermentation and cecal microbiota are prolonged, and further research is warranted to determine their use as a gut health promoter in poultry.

Color, Starch Digestibility, and In Vitro Fermentation of Roasted Highland Barley Flour with Different Fractions.

Highland barley (HB) is commonly milled into flour for direct consumption or further processed with other food formulations. Nevertheless, the association between milling and HB flour properties remains lacking. This work studied the effect of particle sizes (coarse, 250–500 μm; medium, 150–250 μm; fine, <150 μm) on physicochemical and nutritional properties of raw and sand-roasted HB flour. Gelatinization enthalpy decreased with increasing particle sizes of raw HB flour, while no endothermic transitions were observed in sand-roasted flour. Sand roasting destroyed starch granules and decreased short-range molecular order. Starch digestibility increased while total short-chain fatty acids (SCFAs) production decreased with decreasing particle sizes in all samples. The relative crystallinity of sand-roasted HB flour decreased by 80–88% compared with raw samples. Sand roasting raised in vitro starch digestibility, while total SCFAs during in vitro fecal fermentation decreased. Sand-roasted HB flour with particle sizes <150 μm had the highest starch digestibility (94.0%) but the lowest production of total SCFAs (1.89–2.24 mM). Pearson’s correlation analysis confirmed the relationship between the nutritional qualities of HB flour and milling.

Sources of Dietary Fiber Affect the SCFA Production and Absorption in the Hindgut of Growing Pigs.

Effects of different dietary fiber (DF) sources on short-chain fatty acids (SCFA) production and absorption in the hindgut of growing pigs were studied by an in vivo–vitro (ileal cannulated pigs and fecal inoculum-based fermentation) method. Thirty-six cannulated pigs (body weight: 48.5 ± 2.1 kg) were randomly allocated to 6 treatments containing the same DF content (16.5%), with either wheat bran (WB), corn bran (CB), sugar beet pulp (SBP), oat bran (OB), soybean hulls (SH), or rice bran (RB) as DF sources. Pigs were allowed 15 days for diet adaptation, and then, fresh ileal digesta and feces were collected to determine SCFA concentration which was normalized for food dry matter intake (DMI) and the hindgut DF fermentability. Fecal microbiota was inoculated into the freeze-dried ileal digesta samples to predict the ability of SCFA production and absorption in the hindgut by in vitro fermentation. The SH group had the largest concentration of total SCFA and propionate in ileal digesta and fecal samples of growing pigs (p < 0.05). Nonetheless, the predicted acetate, total SCFA production, absorption in the SBP group were the highest (p < 0.01), but the lowest in the OB group (p < 0.01) among all groups. Even SBP and OB group had a similar ratio of soluble DF (SDF) to insoluble DF (IDF). The CB group had high determined ileal and fecal butyrate concentration but the lowest butyrate production and absorption in the hindgut (p < 0.01). Overall, the source of DF had a great impact on the hindgut SCFA production and absorption, and SBP fiber had a great potential to increase hindgut SCFA production and absorption.

Bioaccessibility and movement of phenolic compounds from tomato (Solanum lycopersicum) during in vitro gastrointestinal digestion and colonic fermentation.

Tomatoes (Solanum lycopersicum) are highly involved in diets consumed worldwide, and are rich in bioactive compounds including phenolics, carotenoids and vitamins. In this study, four different varieties of fresh tomato pulp (Oxheart, Green Zebra, Kumato and Roma) were used to estimate the bioaccessibility of target phenolic compounds during in vitro gastrointestinal digestion and colonic fermentation, and to determine their antioxidant capacity. The production of short chain fatty acids (SCFAs) was also estimated during colonic fermentation. Among these, Roma displayed relatively higher total phenolic content (TPC) and free radical scavenging (2,2'-diphenyl-1-picrylhydrazyl (DPPH) assay) values after gastrointestinal digestion of 0.31 mg gallic acid equivalents (GAE) per g and 0.12 mg Trolox equivalents (TE) per g. Kumato exhibited the highest total flavonoid content (TFC) of 2.47 mg quercetin equivalents (QE) per g after 8 hours of colonic fermentation. Oxheart and Roma showed similar ferric reducing antioxidant power (FRAP) values of around 4.30 mg QE per g after 4 hours of faecal reaction. Catechin was the most bioaccessible phenolic compound in all fresh tomatoes, and could be completely decomposed after intestinal digestion, whereas the release of some bonded phenolic compounds required the action of gut microflora. Kumato and Green Zebra showed higher production of individual and total SCFAs for 16 hours of fermentation, which would provide more gut health benefits.

Microbial fermentation of Fossence™, a short-chain fructo-oligosaccharide, under simulated human proximal colonic condition and assessment of its prebiotic effects-a pilot study.

A short-chain fructo-oligosaccharide (sc-FOS) was tested in a simulator of the human gut microbial ecosystem (SHIME) in vitro model to quantify its prebiotic effects according to Prebiotic Index (PI) and Measure of prebiotic effect (MPE) equations. FossenceTM, (sc-FOS, 0.5%) was fermented in a simulated human proximal colonic condition, using a fecal inoculum from a healthy individual. We analysed the pH reduction, substrate utilization, lactate and short-chain fatty acid (SCFA) production and microbial community modulation. Microbial fermentation of sc-FOS strongly reduced the media pH indicating the production of lactate and SCFA with accumulation of lactate and enhanced levels of acetate (34.38±0.38 mM),propionate(20.93±0.56 mM)andbutyrate(4.93±0.03 mM)comparedto18.46±0.20 mM,6.24±0.10 mMand3.3±0.06 mMintheblank, respectively. Total SCFA production in test media was 61.91±0.87 mMcompared to 33.65±0.36 mMin blank and the contribution of free-sugars present in sc-FOS to SCFAs was negligible. Modulation of the microbial community was analysed through 16S rRNA sequencing and we found that sc-FOS greatly stimulated the beneficial bacteria such as Bifidobacteria and Lactobacillus. We report the PI and MPE values for FossenceTM, as 14.9 and 0.01 respectively at the end of 24 h,whichisanindicator of a strong prebiotic effect.

Nutrient load acts as a driver of gut microbiota load, community composition and metabolic functionality in the simulator of the human intestinal microbial ecosystem.

A recently introduced quantitative framework for gut microbiota analysis indicated that microbial load alterations can be linked to various diseases, making it essential to pinpoint its determinants. We identified nutrient load as a main driver of the quantitative microbial community composition and functionality in vitro by stepwise decreasing standardized feed concentrations from 100% to 33%, 20% and 10% in 5-day intervals. While the proportional composition and metabolic profile were mainly determined by the inter-individual variability (35% and 41%), nutrient load accounted for 58%, 23% and 65% of the observed variation in the microbial load, quantitative composition and net daily metabolite production, respectively. After the 10-fold nutrient reduction, the microbial load decreased by 79.72± 9% and 82.96± 1.66% in the proximal and distal colon, respectively, while the net total short-chain fatty acid production dropped by 79.42± 4.42% and 84.58± 2.42%, respectively. The majority of microbial taxa quantitatively decreased, whereas a select group of nutritional specialists, such as Akkermansia muciniphila and Bilophila wadsworthia, and a number of opportunistic pathogens remained unaffected. This shows that nutrient load is an important driver of the human gut microbiome and should be considered in future in vitro and in vivo dietary research.

Antioxidant activity and short-chain fatty acid production of lactic acid bacteria isolated from Korean individuals and fermented foods

Compounds of the cell walls of heat-killed lactic acid bacteria show immunomodulatory properties which boost immunological systems, and are used ad postbiotics (paraprobiotics). In this study, we used 17 different heat-killed isolates as postbiotics and evaluated their anti-inflammatory potential on the expression of proinflammatory mediators and cellular signaling pathways of murine macrophage, RAW 264.7 cells. Bifidobacterium bifidum MG731 showed the high 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activity (90.6%), followed by Bifidobacterium lactis MG741 (59.6%). The Bi. lactis MG741 showed the high ABTS free radical scavenging activity (99.5%), followed by Lactobacillus plantarum MG989 (98.9%), Lactobacillus salivarius MG242 (97.1%), and Bi. bifidum MG731 (96.1%). In addition, Bi. bifidum MG731 showed the lowest nitric oxide production (4.28 µM), followed by B. lactis MG741 (10.80 µM), L. salivarius MG242 (14.60 µM), and L. plantarum MG989 (19.60 µM). The selected strains showed a decreased nitric oxide production via downregulation of inducible nitric oxide synthase and cyclooxygenase 2, which were upregulated via LPS-stimulated RAW 264.7 macrophages. Short-chain fatty acids (SCFA) including acetic, propionic, and butyric acid were produced by four strains. The Bi. bifidum MG731 showed total SCFAs production (4998.6 µg/g), Bi. lactis MG741 (2613.9 µg/g), L. salivarius MG242 (1456.1 µg/g), and L. plantarum MG989 (630.2 µg/g). These results indicated that the various selected strains may possess an anti-inflammatory potential and provide a molecular basis for the development of functional probiotics.

Ammonia fiber expansion increases in vitro digestibility and fermentability of corn distillers dried grains with solubles with or without carbohydrases

A significant portion of gross energy (GE) in corn distillers dried grains with solubles (DDGS) is indigestible by non-ruminants, and the impact of ammonia fiber expansion (AFEX) on improving GE digestibility is unknown. The objective of this study was to determine the effect of AFEX and enzymes on crystallinity index and neutral detergent fiber (aNDF) degradation in DDGS using a porcine in vitro fermentation system. Two sources of DDGS samples (ICM and POET) were pretreated with liquid ammonia at 100 °C and 2069 kPa for 30 min. The DDGS samples with or without AFEX pretreatment were predigested with pepsin and pancreatin with or without carbohydrases (1500 U/g xylanase, 1100 U/g β-glucanase, 110 U/g mannanase, and 35 U/g galactosidase). Residues were then subjected to in vitro fermentation (directly inoculating DDGS samples with fecal inocula) and accumulated gas production recorded up to 72 h. Concentration of short-chain fatty acids (SCFA) were measured in the fermented solution. On a dry matter (DM) basis, non-protein nitrogen was increased by 20.8 g/kg, and aNDF was decreased by 146 g/kg in corn DDGS after AFEX pretreatment. Pretreatment with AFEX decreased the crystallinity index of fiber in DDGS from about 15 to 0 %, increased (P < 0.01) fermentability of DM and aNDF, and decreased (P < 0.01) unfermented aNDF content after in vitro fermentation for 12 and 72 h. The AFEX pretreated DDGS had greater (P < 0.05) in vitro digestibility of DM and GE, greater production of acetic acid and total SCFA (DM enzymatic hydrolyzed residue basis), greater in vitro digested GE (GE × IVDGE, GE) and digestible energy (GE + energy derived from SCFA) compared with DDGS without pretreatment. Carbohydrase supplementation increased (P < 0.05) in vitro digestibility of GE and DM in AFEX treated POET but not in ICM DDGS. In conclusion, AFEX pretreatment decreased the crystallinity index of fiber and improved the in vitro digestibility and fermentability of corn DDGS.

Lactobacillus rhamnosus from human breast milk ameliorates ulcerative colitis in mice via gut microbiota modulation.

Gut microbiota imbalance is one of the major causes of ulcerative colitis (UC). L. rhamnosus SHA113 (LRS), a strain isolated from healthy human milk, influences the regulation of gut flora. This study aims to determine whether this strain can ameliorate UC by modulating gut microbiota. Mouse models of UC were established using C57BL/6Cnc mice with intragastric administration of 3.0% (w/v) dextran sodium sulfate (DSS). LRS was used to treat the mouse models of UC with 109 cfu mL-1 cell suspension via intragastric administration. To verify the effect of gut microbiota on UC, fecal microbiota collected from the mice after the treatment with LRS were also used to treat the UC mouse models (FMT). The severity of UC was evaluated based on body weight, colon length, disease activity index (DAI), and hematoxylin-eosin staining. The microbial composition was analyzed by 16S rRNA sequencing. The mRNA expression levels of cytokines, mucins, tight junction proteins, and antimicrobial peptides in the gastrointestinal tract were detected by quantitative real-time polymerase chain reaction. The short-chain fatty acid (SCFAs) in the cecal contents of all mice were quantitatively detected by gas chromatography and mass spectrometry. Both LRS and FMT exerted excellent therapeutic effects on UC, as evidenced by the reduction in body weight loss, colon length, and colon structural integrity, as well as the increase in the DAI (disease activity index). LRS and FMT treatments showed similar effects: (1) an increase of total SCFA production in the cecal contents and the abundance of gut microbial diversity and flora composition; (2) decreases in two genera (Parabacteroides and Escherichia/Shigella) related to the DAI and the enhancement of SCFAs and IL-10 positively related genera in the gut microbiota (Bilophila, Roseburia, Akkermansia, and Bifidobacterium); (3) downregulation of the expression of tumor necrosis factor-α, interleukin IL-6, and IL-1β, and upregulation of the expression of the anti-inflammatory cytokine IL-10; and (4) upregulation of the expression of mucins (Muc1-4) and tight junction protein ZO-1. Overall, L. rhamnosus SHA113 relieves UC via the regulation of gut microbiota: increases in SCFA-producing genera and decreases in UC-related genera. In addition, a single strain is sufficient to induce a significant change in the gut microbiota and exert therapeutic effects on UC.

Active phase prebiotic feeding alters gut microbiota, induces weight-independent alleviation of hepatic steatosis and serum cholesterol in high-fat diet-fed mice

Growing evidence suggests that prebiotics may induce weight loss and alleviate non-alcoholic fatty liver disease (NAFLD) via modulation of the gut microbiota. However, key members of the gut microbiota that may mediate the beneficial effects of prebiotics remain elusive. Here, we find that restricted prebiotic feeding during active phase (HF-ARP) induced weight-independent alleviation of liver steatosis and reduced serum cholesterol in high-fat diet (HF) fed mice more significantly than unrestricted feeding (HF-UP). HF-ARP mice also showed concomitantly altered gut microbiota structure that was different from HF-UP group along with significantly increased production of total short-chain fatty-acids (SCFAs). Amplicon sequence variants (ASVs) were clustered into co-abundant groups (CAGs) as potential functional groups that may respond distinctively to prebiotic consumption and prebiotic feeding regime. Prebiotic feeding induces significant alterations in CAG abundances by day 7. Eight of 32 CAGs were promoted by prebiotics, including CAG17 with the most abundant ASV from Parabacteroides, CAG22 with Bacteroides thetaiotamicron and CAG32 with Fecalibaculum and Akkermansia. Among the prebiotic-promoted CAGs, CAG20 with ASVs from Lachnospiraceae and CAG21 with ASVs from Bifidobacterium and Lachnospiraceae were significantly enhanced in HF-ARP compared to HF-UP. Moreover, most of the prebiotic-promoted CAGs were also significantly associated with improvements in hepatic steatosis, reduction in serum cholesterol and increased cecal propionate production. Together, these results suggest that the impact of prebiotics on weight-independent alleviation of liver steatosis and cholesterol-lowering effect can be optimized by restricting prebiotic intake to active phase and is associated with a distinct change of gut microbiota with increased SCFA production.

Heat Stress Increases In Vitro Hindgut Fermentation of Distinct Substrates in Iberian Pigs.

Simple SummaryHeat stress is a major concern in pig production in summer, as pigs have a limited number of functional sweat glands to transfer body heat. Above 25 °C pigs are out of their comfort zone and mechanisms such as decreasing feed intake or diverting blood from the internal organs to the skin are triggered. Intestinal microbiota is also affected by high ambient temperature but the consequences on fermentation capacity are poorly known. Short-chain fatty acids are the end-products of bacterial metabolism of carbohydrates and protein mainly in the hindgut and, in addition to being a source of energy, they have beneficial effects on immune status and health. An understanding of the effects of heat stress on intestinal fermentation could help to develop strategies mitigating intestinal disorders. We used an in vitro method to assess gas and short-chain fatty acid production, utilizing as inoculum feces from Iberian pigs fed a commercial diet for 28 days under neutral (20 °C) or heat stress (30 °C) conditions. Four substrates with dissimilar fermentation characteristics were incubated in vitro with fecal inoculum for 24 h. Chronic heat stress increased in vitro production of short-chain fatty acids, suggesting a modification of intestinal microbiota activity.Heat stress reduces the feed intake and growth of pigs. We hypothesized that heat stress affects the intestinal fermentation capacity of pigs. Sixteen Iberian pigs (44 ± 1.0 kg) were randomly assigned to one of two treatments (eight pigs/treatment) for 4 weeks—heat stress (HS; 30 °C) ad libitum or thermoneutral (TN; 20 °C) pair feeding. Frozen rectum contents were used as inocula for 24 h in vitro incubations in which a mixture of starches, citrus pectin, inulin from chicory, and cellulose were the substrates. Cellulose was poorly degraded, whereas pectin and the mixture of starches were the most fermentable substrates according to total short-chain fatty acid (SCFA) production. The mixture of starches and inulin produced the greatest amount of gas. For all substrates, heat stress enhanced gas production (8%, p = 0.001), total SCFA production (16%, p = 0.001), and the production of acetate and propionate (12% and 42%, respectively; p = 0.001). The increased isoacid production (33%, p = 0.001) and ammonia concentration (12%, p = 0.001) may indicate protein fermentation under heat stress. In conclusion, the in vitro intestinal fermentation capacity of pigs under heat stress was increased compared to thermoneutral conditions, which may indicate an adaptive response to heat stress.

Pretreatment of Rapeseed Meal Increases Its Recalcitrant Fiber Fermentation and Alters the Microbial Community in an in vitro Model of Swine Large Intestine.

The aim of current study was to investigate in an in vitro study how enzymatic and chemical pretreated rapeseed meal (RSM) influences the fiber fermentation and microbial community in the swine large intestine. RSM was processed enzymatically by a cellulase (CELL), two pectinases (PECT), or chemically by an alkaline (ALK) treatment. 16S rRNA gene sequencing data was performed to evaluate changes in the gut microbiota composition, whereas short-chain fatty acid (SCFA) production (ion-chromatography) and non-starch polysaccharides (NSP) composition (using monoclonal antibodies; mAbs) were used to assess fiber degradation. The results showed that ALK, CELL, PECT1, and PECT2 changed microbial community composition, increased the predicted abundance of microbial fiber-degrading enzymes and pathways, and increased acetic acid, propionic acid, butyric acid, and total SCFA production. The increased microbial genera positively correlated with SCFA production. Monoclonal antibody analyses showed that the cell wall polysaccharide structures of RSM shifted after ALK, CELL, PECT1, and PECT2 treatment. The degradation of NSP during the fermentation period was dynamic, and not continuous based on the epitope recognition by mAbs. This study provides the first detailed analysis of changes in the swine intestinal microbiota due to RSM modified by ALK, CELL, PECT1, and PECT2, which altered the microbial community structure, shifted the predicted functional metagenomic profile and subsequently increased total SCFA production. Our findings that ALK, CELL, PECT1, and PECT2 increased fiber degradability in RSM could help guide feed additive strategies to improve efficiency and productivity in swine industry. The current study gave insight into how enzymatic treatment of feed can alter microbial communities, which provides good opportunity to develop novel carbohydrase treatments, particularly in swine feed.