In vitro fermentability of dextran, oligodextran and maltodextrin by human gut bacteria

Anaerobic batch culture fermenters were used for a preliminary screening of the in vitro utilization by human gut microflora of dextran and novel oligodextrans (I, II and III) produced in the University of Reading (UK). Glucose and fructooligosaccharides (FOS) were used as reference carbohydrates. As expected, FOS acted as a good prebiotic in that it selectively increased numbers of bifidobacteria in the early stages of the fermentation. Dextran and oligodextrans each resulted in an enrichment of bifidobacteria in the batch cultures, with high levels of persistence up to 48 h. They also produced elevated levels of butyrate ranging from 5 to 14.85 mmol/l. To more effectively simulate conditions that prevail in different regions of the large intestine, a three-stage continuous culture cascade system was used to study further the fermentation of dextran, a low-molecular-mass oligodextran (IV) and maltodextrin. Oligodextran IV was shown to be the best substrate for bifidobacteria and lactobacilli with steady-state populations of bifidobacteria and lactobacilli being higher in all three vessels of the gut model than the respective populations resulting from dextran and maltodextrin. A maximum difference of 1.9 log was observed in vessel 1 for both bifidobacteria and lactobacilli in the case of dextran fermentation, with 1.4 log and 0.8 log in vessel 3 were the maximum differences for bifidobacteria and lactobacilli when maltodextrin was used as the carbohydrate source. Moreover, dextran and oligodextran appeared to stimulate butyrate production, with a maximum production up to 25.39 mmol/l in vessel 3 when fermenting dextran, followed by 21.70 mmol/l in the case of oligodextran IV and only 12.64 mmol/l in the case of maltodextrin.     

In vitro fermentation of gum acacia – impact on the faecal microbiota

Interest in the consumption of gum acacia (GA) has been associated with beneficial health effects, which may be mediated in part by prebiotic activity. Two doses of GA and fructooligosaccharide (FOS) (1 and 2%) were tested for their efficacy over 48?h in pH- and temperature-controlled anaerobic batch cultures inoculated with human faeces. Samples were taken after 0, 5, 10, 24 and 48?h of fermentation. The selective effects of GA (increases in Bifidobacterium spp. and Lactobacillus spp.) were similar to those of the known prebiotic FOS. The 1% dose of substrates showed more enhanced selectivity compared to the 2% dose. The fermentation of GA also led to SCFA production, specifically increased acetate after 10, 24 and 48?h of fermentation, propionate after 48?h and butyrate after 24 and 48?h. In addition, FOS led to significant increase in the main SCFAs. These results suggest that GA displays potential prebiotic properties.     

Carbohydrates blended with polydextrose lower gas production and short-chain fatty acid production in an in vitro system

Maximizing health benefits of prebiotics, while limiting negative side effects, is of importance to the food industry. This study examined several oligosaccharides and their blends in an in vitro fermentation model. Substrates included medium- and long-chain fructooligosaccharides (FOS), oligofructose-enriched inulin, galactooligosaccharide, polydextrose (POL), and 50:50 substrate blends. Substrates and blends were fermented in vitro using human fecal inoculum, and fermentation characteristics were quantified at 0, 4, 8, and 12 hours. We hypothesized that mixtures of short- and long-chain oligosaccharides would generate less gas than do short-chain oligosaccharides and modulate gut microflora to a greater extent than do long-chain oligosaccharides. Carbohydrates blended with POL had decreased (P < .01) total gas volume and H₂ produced after 4, 8, and 12 hours of fermentation compared with individual carbohydrates. Mixing of 2 oligofructose-enriched inulin products led to less (P < .05) gas produced and a slower (P < .05) rate of production. When mixed with POL, all carbohydrates tested in the present study produced less total short-chain fatty acids (P < .04) and butyrate (P < .0001) after 12 hours of in vitro fermentation, compared with individual carbohydrates. The bifidogenic effect of medium-chain FOS and oligofructose-enriched inulin after 12 hours of in vitro fermentation was lower (P < .05) when mixed with POL. Mixing the pure carbohydrates with galactooligosaccharide increased (P < .05) bifidobacteria counts measured after 12 hours of in vitro fermentation, except when mixed with medium-chain FOS. In general, when mixed with POL, all carbohydrates had lower gas production, gas production rates, butyrate and total short-chain fatty acid production, and bifidobacteria counts than when fermented alone for 12 hours.     

Fructooligosaccharides and Lactobacillus acidophilus modify gut microbial populations,total tract nutrient digestibilities and fecal protein catabolite concentrations in healthy adult dogs

The objective of this research was to determine whether fructooligosaccharides (FOS) and (or) Lactobacillus acidophilus (LAC) affected concentrations of gut microbial populations, fermentative end products and nutrient digestibilities in healthy adult dogs. Two experiments were performed using 40 adult dogs (20 dogs/experiment). Dogs in each experiment were randomly assigned to one of 4 treatments. Twice daily, treatments were given orally via gelatin capsules: 1) 2 g sucrose + 80 mg cellulose; 2) 2 g FOS + 80 mg cellulose; 3) 2 g sucrose + 1 x 10(9) colony forming units (cfu) LAC; or 4) 2 g FOS + 1 x 10(9) cfu LAC. Data were analyzed by the General Linear Models procedure of SAS. In Experiment 1, FOS resulted in lower (P = 0.08) Clostridium perfringens and greater fecal butyrate (P = 0.06) and lactate (P < 0.05) concentrations. In Experiment 2, FOS supplementation increased (P < 0.05) bifidobacteria, increased lactobacilli (P = 0.08), increased fecal lactate (P = 0.06) and butyrate (P < 0.05), and decreased (P < 0.05) fecal ammonia, isobutyrate, isovalerate and total branched-chain fatty acid concentrations. Dogs fed LAC had the highest fecal concentrations of hydrogen sulfide and methanethiol in Experiment 1 and dimethyl sulfide in Experiment 2, whereas dogs fed FOS had the lowest concentrations of these compounds. Overall, FOS appeared to enhance indices of gut health by positively altering gut microbial ecology and fecal protein catabolites, whereas LAC was more effective when fed in combination with FOS rather than fed alone.     

Dietary fructo-oligosaccharides in healthy adults do not negatively affect faecal cytotoxicity: a randomised, double-blind, placebo-controlled crossover trial

Fructo-oligosaccharides (FOS) are widely used in commercial food products. Most studies on FOS concern the health benefits, but some negative effects were recently reported concerning the faecal cytotoxicity and excretion of mucin-type oligosaccharides in combination with a Ca-restricted diet. The present study was performed to investigate whether these effects of FOS are observed in adults consuming a regular diet unrestricted in Ca. The study was a randomised, double-blind, placebo-controlled crossover trial, involving eleven healthy adults, who consumed 25-30 g FOS or maltodextrin (control) in a random order for 2 weeks in addition to their regular diet. Stools were collected for analysis of pH and SCFA (as markers of fermentation), for the assessment of faecal water cytotoxicity, and for the analysis of alkaline phosphatase activity (as a marker of epithelial cell turnover) and O-linked oligosaccharides (to estimate the excretion of mucin-type oligosaccharides). FOS consumption significantly altered bacterial fermentation (increased percentage of acetate, decreased percentage of butyrate) and tended to decrease stool pH. Furthermore, FOS consumption resulted in a significantly higher stool frequency and in significantly more complaints of flatulence. No significant differences between the control and FOS period were observed in the mean cytotoxicity of faecal water (37.5 (SEM 6.9)% v. 18.5 (SEM 6.9)%; P=0.084), in mean alkaline phosphatase activity (27.7 (SEM 2.9) v. 24.6 (SEM 3.2) U/g dry faeces; P=0.496) or in the mean excretion of mucin-type oligosaccharides (49.9 (sem 4.0) v. 53.5 (SEM 4.3) mg/g dry faeces; P=0.553). We conclude that dietary FOS in a dose up to 25-30 g/d altered the bacterial fermentation pattern but did not affect faecal cytotoxicity or the faecal concentration of mucin-type oligosaccharides in human adults consuming a regular diet.     

Supplemental fructooligosaccharides and mannanoligosaccharides influence immune function, ileal and total tract nutrient digestibilities, microbial populations and concentrations of protein catabolites in the large bowel of dogs

The goal of this study was to examine whether supplemental fructooligosaccharides (FOS) and (or) mannanoligosaccharides (MOS) influenced indices of gut health of dogs. Adult female dogs (n = 4) surgically fitted with ileal cannulas were fed a dry, extruded, kibble diet twice daily. At each feeding, the following treatments were administered: 1) Control (no FOS or MOS); 2) 1 g FOS; 3) 1 g MOS; or 4) 1 g FOS + 1 g MOS. Fecal, ileal and blood samples were collected during the last 4 d of each 14-d period to measure protein catabolite concentrations, microbial populations, immune characteristics and nutrient digestibilities. Treatment means were compared using preplanned orthogonal contrasts. Dogs supplemented with MOS had lower (P = 0.05) fecal total aerobes and tended to have greater (P = 0.13) Lactobacillus populations. Ileal immunoglobulin (Ig) A concentrations were greater (P = 0.05) in dogs supplemented with FOS + MOS vs. control. Lymphocytes (% of total white blood cells) were greater (P < 0.05) in dogs supplemented with MOS. Serum IgA concentrations also tended (P = 0.13) to be greater in dogs supplemented with MOS. Dogs supplemented with FOS and FOS + MOS had lower (P < 0.05) fecal total indole and phenol concentrations. Dogs supplemented with MOS tended to have lower ileal DM (P = 0.149) and OM (P = 0.146) digestibilities vs. control. Results of this study suggest that dietary supplementation of FOS and MOS may have beneficial effects on colonic health and immune status of dogs.     

Dietary fructo-oligosaccharides dose-dependently increase translocation of salmonella in rats

Prebiotics, such as fructo-oligosaccharides (FOS), stimulate the protective gut microflora, resulting in an increased production of organic acids. This may result in increased luminal killing of acid-sensitive pathogens. However, host defense against invasive pathogens, like salmonella, also depends on the barrier function of the intestinal mucosa. Rapid fermentation of prebiotics leading to high concentrations of organic acids may impair the barrier function. Therefore, we determined the dose-dependent effect of dietary FOS on the resistance of rats to Salmonella enteritidis. Male Wistar rats were fed restricted quantities of a "humanized" purified diet supplemented with 0, 3 or 6 g/100 g of FOS (n = 7 in the 6% FOS group and n = 8 in the other diet groups). After an adaptation period of 2 wk, rats were orally infected with 1.7 x 10(10) colony-forming units of S. enteritidis. Supplement-induced changes in the intestinal microflora and fecal cation excretion were determined before and after infection. Cytotoxicity of fecal water was determined with an in vitro bioassay, and fecal mucins were quantified fluorimetrically. Colonization of S. enteritidis was determined by quantification of salmonella in cecal contents and mucosa. Translocation of S. enteritidis was quantified by analysis of urinary nitric oxide metabolites in time. Before infection, FOS decreased cecal and fecal pH, increased fecal lactic acid concentration and increased bifidobacteria and enterobacteria. FOS also increased cytotoxicity of fecal water and fecal mucin excretion, indicating mucosal irritation. Remarkably, FOS dose-dependently increased salmonella numbers in cecal contents and mucosa and caused a major increase in infection-induced diarrhea. In addition, FOS enhanced translocation of salmonella. Thus, in contrast to most expectations, FOS dose-dependently impairs the resistance to salmonella infection in rats. These results await verification by other controlled animal and human studies.     

Fructo-oligosaccharides enhance the mineral absorption and counteract the adverse effects of phytic acid in mice

ObjectiveWe explored the effects of fructo-oligosaccharides (FOS) and phytic acid (PA) on the absorption of minerals and their interaction.MethodsA 3 × 2 factorial experiment was designed to evaluate the effects of FOS (in the presence or absence of PA) on the apparent absorption rate of minerals and the mineral status (plasma, hepatic, and bone) in mice. Sixty Kun-Ming mice were randomized into six groups: basal diet group; basal diet + 1% PA group (PA); basal diet + 0.8 g/kg of body weight FOS group (FOS1); FOS1 + 1% PA group (FOS1 + PA); basal diet + 2.5 g/kg of body weight group (FOS2); and FOS2 + 1% PA group (FOS2 + PA). The mice received FOS by gavage for consecutive 4 wk, and the PA was added in the diet. The mice were housed individually in the last week. The food intake was recorded and the feces were collected for calculation of the apparent absorption rate. Then the mice were sacrificed, the ceca were removed and weighed, and the cecum contents were used for the detection of pH and short-chain fatty acids. The blood, liver, and the left femur were collected for the measurement of the minerals.ResultsFOS supplementation resulted in the enlargement of the cecum and increased cecal acidification (P < 0.01). In addition, FOS effectively boosted the apparent absorption rate of calcium (FOS1, +7%; FOS2, +9%, P < 0.05), magnesium (FOS1, +26%; FOS2, +19%, P < 0.05), and iron (FOS1, +17%; FOS2, +22%, P < 0.05), and restored the PA-impaired magnesium and iron apparent absorption rates (P < 0.01). In addition, FOS significantly increased hepatic zinc levels (P < 0.01) and femoral magnesium levels (P < 0.01).ConclusionThese data indicate that FOS effectively enhances the mineral apparent absorption rate and counteracts the deleterious effects of PA.     

Glucose-based oligosaccharides exhibit different in vitro fermentation patterns and affect in vivo apparent nutrient digestibility and microbial populations in dogs

To evaluate the potential of indigestible oligosaccharides (OS) to serve as "dietary fiber-like" ingredients, it is necessary to determine their extent of indigestibility. In vitro fermentation characteristics of two novel OS, alpha-glucooligosaccharides (GOS) and a maltodextrin-like OS (MD), were compared to those of fructooligosaccharides (FOS), gum arabic (GA), guar gum (GG) and guar hydrolysate (GH). Total short-chain fatty acid (SCFA) production (micromol/g dry matter) as a result of MD fermentation was higher initially compared with GA (P<0.01), but GA was more extensively fermented at 24 h (P<0.01). Total SCFA production for GOS was similar to that for FOS, GG, GH and GA. In the second experiment, GOS and MD were added at 6% to an enteral formula control diet (Control) and fed to ileal-cannulated dogs in a 3x3 replicated Latin-square design. Ileal digestibility of glucose was lower (P<0.05) and carbohydrate (CHO) numerically lower (P = 0.08) for both GOS and MD compared with the Control. Total tract digestibility of CHO and glucose was lower only for MD (P<0.01) compared with the Control. Total fecal weights were higher (P<0.01) for both GOS and MD treatments. Fecal concentration of bifidobacteria was numerically increased by GOS and MD supplementation (P = 0.13 and 0.23, respectively). Thus, GOS and MD are indigestible yet fermentable OS, and may act as "dietary fiber-like" ingredients.     

Effect of a prebiotic mixture on intestinal comfort and general wellbeing in health

Specific carbohydrates, i.e. prebiotics such as fructo-oligosaccharide (FOS), are not digested in the small intestine but fermented in the colon. Besides beneficial health effects of an enhanced bifidobacteria population, intestinal gas production resulting from fermentation can induce abdominal symptoms. Partial replacement with slowly fermented acacia gum may attenuate side effects. The aim was to compare the effects of FOS with those of a prebiotic mixture (50 % FOS and 50 % acacia gum; BLEND) and a rapidly absorbed carbohydrate (maltodextrin) on general intestinal wellbeing, abdominal comfort and anorectal sensory function. Twenty volunteers (eight male and twelve female; age 20-37 years) completed this double-blind, randomised study with two cycles of a 2-week run-in phase (10 g maltodextrin) followed by 5 weeks of 10 g FOS or BLEND once daily, separated by a 4-week wash-out interval. Abdominal symptoms and general wellbeing were documented by telephone interview or Internet twice weekly. Rectal sensations were assessed by a visual analogue scale during a rectal barostat test after FOS and BLEND treatment. Both FOS and BLEND induced more side effects than maltodextrin. Belching was more pronounced under FOS compared with BLEND (P = 0.09 for females; P = 0.01 for males), and for self-reported general wellbeing strong sex differences were reported (P = 0.002). Urgency scores during rectal barostat were higher with FOS than BLEND (P = 0.01). Faced with a growing range of supplemented food products, consumers may benefit from prebiotic mixtures which cause fewer abdominal side effects. Sex differences must be taken in consideration when food supplements are used.     

A randomized controlled trial of phytoestrogen supplementation,growth and bone turnover in adolescent males

OBJECTIVE: To assess the effect of phytoestrogens on bone turnover and growth in adolescent boys. DESIGN: Randomized double-blind placebo-controlled trial. SETTING: Single school in northwest Tasmania. PARTICIPANTS: Adolescent boys (treatment n=69, placebo n=59, mean age 16.8 y). INTERVENTIONS: Six weeks of isoflavone supplementation (Novasoy, 50 mg daily of isoflavone equivalents). Bone turnover markers (bone specific alkaline phosphatase (BAP) and pyridinoline creatinine ratio (PYR)) were measured at baseline and follow-up. RESULTS: Despite marked increases in urinary genistein and daidzein in the treatment arm (both P<0.001), there were no significant differences in BAP, PYR or short-term height or weight change. This applied to both intention-to-treat and per protocol analysis. Neither was there a significant correlation between urinary genistein and daidzein levels and BAP or PYR. CONCLUSIONS: Phytoestrogen supplementation to the level of usual Japanese dietary intake has no measurable effect on bone turnover in adolescent boys. Longer-term studies of bone density may be desirable but it is unlikely that there will be a large effect in either girls or boys given the lower endogenous oestrogen levels in boys.     

Dietary fructooligosaccharides modify intestinal bioavailability of a single dose of genistein and daidzein and affect their urinary excretion and kinetics in blood of rats

The influence of dietary fructooligosaccharides (FOS) on bioavailability of genistein and daidzein in rats was estimated by measuring their concentrations in plasma collected from three different veins and in urine after a single intragastric administration of isoflavone conjugates. Sprague-Dawley male rats (6 wk old, n = 22) were fed a purified control (AIN-93G) diet or a FOS diet (AIN-93G + 5% FOS) for 7 d. A single dose of soy isoflavone conjugates, i.e., 8.5 mg as genistein and 33 mg as daidzein/kg body, was administered via a stomach tube at d 5. Blood samples were collected after administration via catheters in the portal and central veins and by puncture of the tail vein. The isoflavones in plasma and urine were analyzed by time-resolved fluoroimmunoassay. The genistein concentration in the portal blood increased rapidly, reaching a peak of 3.5 micromol/L in both groups at 1 h after administration. The concentrations in the central and tail venous blood were approximately half of those in the portal blood. In the FOS-fed group, both genistein and daidzein remained detectable at 24 and 48 h in the tail venous plasma. The urinary excretion of both isoflavones in the 24- to 48-h period after administration was significantly higher in the FOS-fed group than in the control group. The difference between the portal and central veins indicated hepatic uptake, probably leading to conjugation of aglycones and excretion into bile. FOS modified the absorption and enterohepatic recirculation of isoflavones.     

Effects of infant formula containing a mixture of galacto- and fructo-oligosaccharides or viable Bifidobacterium animalis on the intestinal microflora during the first 4 months of life

Adding prebiotics or probiotics to infant formula to improve the intestinal flora of formula-fed infants is considered to be a major innovation. Several companies have brought relevant formulations onto the market. However, comparative data on the effects of pre- and probiotics on the intestinal microflora of infants are not available. The present study aimed to compare the effects of infant formula containing a mixture of galacto- and fructo-oligosaccharides or viable Bifidobacterium animalis on the composition and metabolic activity of the intestinal microflora. Before birth, infants were randomised and double blindly allocated to one of three formulas. The prebiotic (GOS/FOS) group (n 19) received regular infant formula supplemented with a mixture of galacto-oligosaccharides and fructo-oligosaccharides (6 g/l). The probiotic (Bb-12) group (n 19) received the same formula supplemented with 6.0x10(10) viable cells of B. animalis per litre. The standard group (n 19) received non-supplemented regular formula. A group of sixty-three breast-fed infants was included as a reference group. Faecal samples were taken at postnatal day 5 and 10, and week 4, 8, 12 and 16. Compared with the groups fed Bb-12 and standard formula, the GOS/FOS formula group showed higher faecal acetate ratio (69.7 % (sem 2.7), 69.9 % (sem 3.9) and 82.2 % (sem 5.3); P<0.05) and lactate concentration (11.3 (sem 7.9), 3.1 (sem 2.3) and 34.7 (sem 10.7) mmol/kg faeces) and lower pH (6.6 (sem 0.2), 7.1 (sem 0.2) and 5.6 (sem 0.2); P<0.05) at 16 weeks. Differences in percentage of bifidobacteria between the GOS/FOS (59.2 % (sem 7.7)), Bb-12 (52.7 % (sem 8.0)) and the standard (51.8 % (sem 6.4)) groups were not statistically significant at 16 weeks. Feeding infants GOS/FOS formula resulted in a similar effect on metabolic activity of the flora as in breast-fed infants. In the Bb-12 group, composition and metabolic activity of the flora were more similar to those of the standard group.     

Quantification of fructans,galacto‐oligosacharides and other short‐chain carbohydrates in processed grains and cereals

Background: Wholegrain grains and cereals contain a wide range of potentially protective factors that are relevant to gastrointestinal health. The prebiotics best studied are fructans [fructooligosaccharides (FOS), inulin] and galactooligosaccharides (GOS). These and other short‐chain carbohydrates can also be poorly absorbed in the small intestine (named fermentable oligo‐, di‐ and monosaccharides and polyols; FODMAPs) and may have important implications for the health of the gut. Methods: In the present study, FODMAPs, including fructose in excess of glucose, FOS (nystose, kestose), GOS (raffinose, stachyose) and sugar polyols (sorbitol, mannitol), were quantified using high‐performance liquid chromatography with an evaporative light scattering detector. Total fructan was quantified using an enzymic hydrolysis method. Results: Fifty‐five commonly consumed grains, breakfast cereals, breads, pulses and biscuits were analysed. Total fructan were the most common short‐chain carbohydrate present in cereal grain products and ranged (g per portion as eaten) from 1.12 g in couscous to 0 g in rice; 0.6 g in dark rye bread to 0.07 g in spelt bread; 0.96 g in wheat‐free muesli to 0.11 g in oats; and 0.81 g in muesli fruit bar to 0.05 g in potato chips. Raffinose and stachyose were most common in pulses. Conclusions: Composition tables including FODMAPs and prebiotics (FOS and GOS) that are naturally present in food will greatly assist research aimed at understanding their physiological role in the gut.     

Restoration of the integrity of rat caeco-colonic mucosa by resistant starch,but not by fructo-oligosaccharides,in dextran sulfate sodium-induced experimental colitis

Butyrate is recognised as efficient in healing colonic inflammation, but cannot be used as a long-term treatment. Dietary fibre that produces a high-butyrate level when fermented represents a promising alternative. We hypothesised that different types of dietary fibre do not have the same efficiency of healing and that this could be correlated to their fermentation characteristics. We compared short-chain fructo-oligosaccharides (FOS) and type 3 resistant starch (RS) in a previously described dextran sulfate sodium (DSS)-induced colitis model. Seventy-two Sprague-Dawley rats received water (control rats) or DSS (50 g DSS/l for 7 d then 30 g DSS/l for 7 (day 7) or 14 (day 14) d). The rats were fed a basal diet (BD), or a FOS or RS diet creating six groups: BD-control, BD-DSS, FOS-control, FOS-DSS, RS-control and RS-DSS. Caeco-colonic inflammatory injuries were assessed macroscopically and histologically. Short-chain fatty acids (SCFA) were quantified in caeco-colon, portal vein and abdominal aorta. At days 7 and 14, caecal and distal macroscopic and histological observations were improved in RS-DSS compared with BD-DSS and also with FOS-DSS rats. Caeco-colonic SCFA were reduced in FOS-DSS and RS-DSS groups compared with healthy controls. The amount of butyrate was higher in the caecum of the RS-DSS rats than in the BD-DSS and FOS-DSS rats, whereas distal butyrate was higher in FOS-DSS rats. Partially explained by higher luminal levels of SCFA, especially butyrate, the healing effect of RS confirms the involvement of some types of dietary fibre in inflammatory bowel disease. Moreover, the ineffectiveness of FOS underlines the importance of the type of dietary substrate.     

Colonic In Vitro Model Assessment of the Prebiotic Potential of Bread Fortified with Polyphenols Rich Olive Fiber

The use of olive pomace could represent an innovative and low-cost strategy to formulate healthier and value-added foods, and bakery products are good candidates for enrichment. In this work, we explored the prebiotic potential of bread enriched with Polyphenol Rich Fiber (PRF), a defatted olive pomace byproduct previously studied in the European Project H2020 EcoProlive. To this aim, after in vitro digestion, the PRF-enriched bread, its standard control, and fructo-oligosaccharides (FOS) underwent distal colonic fermentation using the in vitro colon model MICODE (multi-unit colon gut model). Sampling was done prior, over and after 24 h of fermentation, then metabolomic analysis by Solid Phase Micro Extraction Gas Chromatography Mass Spectrometry (SPME GCMS), 16S-rDNA genomic sequencing of colonic microbiota by MiSeq, and absolute quantification of main bacterial species by qPCR were performed. The results indicated that PRF-enriched bread generated positive effects on the host gut model: (i) surge in eubiosis; (ii) increased abundance of beneficial bacterial groups, such as Bifidobacteriaceae and Lactobacillales; (iii) production of certain bioactive metabolites, such as low organic fatty acids; (iv) reduction in detrimental compounds, such as skatole. Our study not only evidenced the prebiotic role of PRF-enriched bread, thereby paving the road for further use of olive by-products, but also highlighted the potential of the in vitro gut model MICODE in the critical evaluation of functionality of food prototypes as modulators of the gut microbiota.     

Modification of an in vitro model simulating the whole digestive process to investigate cellular endpoints of chemoprevention

In vitro gut fermentation systems are relevant tools to study health benefits of foodstuffs. Most of them are commonly used to investigate the degradation of nutrients or the development of gut flora. Using these models, strong cytotoxic effects of the resulting samples on cultured cells were observed. Hence, the aim of the present study was to develop a modified in vitro fermentation model that simulates the whole digestive tract and generates fermented samples that are suitable for testing in cell culture experiments. Wholemeal wheat flour (wwf) was digested and fermented in vitro with a fermentation model using different ox gall concentrations (41·6 and 0·6?g/l). The resulting fermentation supernatants (fs) were characterised for metabolites and biological effects in HT29 cells. The fermentation of wwf increased chemopreventive SCFA and decreased carcinogenic deoxycholic acid (DCA). The strong cytotoxic effects of the fs, which were partly due to cholic acid and DCA, were diminished by lowering the ox gall concentration, allowing the use of the samples in cell culture experiments. In conclusion, an in vitro digestion model, which can be used to study the effects of foodstuffs on chemoprevention and gut health in colon cells, is introduced and its physiological relevance is demonstrated.     

Promotive effects of non-digestible disaccharides on rat mineral absorption depend on the type of saccharide

OBJECTIVE: We examined the effects of feeding non-digestible disaccharides, difructose anhydride III (DFAIII), maltitol, melibiose and, cellobiose, on calcium, magnesium, and iron absorption in comparison with fructo-oligosaccharide (FOS) in normal and ovariectomized rats. METHODS: In experiment 1, six groups of male Sprague-Dawley rats were fed a control diet (100 g of cellulose/kg of diet), test diets containing 30 g of FOS, or the four non-digestible disaccharides in place of the cellulose in the control diet for 4 wk. In experiment 2, two groups of female Sprague-Dawley rats (sham or ovariectomized) were assigned to one of four subgroups and fed the control or test diet containing FOS, DFAIII, and melibiose for 5 wk. Feces and cecal contents were collected to evaluate mineral absorption and cecal fermentation. RESULTS: In experiment 1, calcium absorption in all the disaccharides groups except the cellobiose group, magnesium absorption in all test diet groups, and iron absorption in the FOS, DFAIII, and melibiose groups were higher than those in the control group. In ovariectomized rats (experiment 2), calcium absorption in the DFAIII and melibiose groups, magnesium absorption in all test diet groups, and iron absorption in the DFAIII group alone were higher than those in the control group. Cecal organic acids were positively and pH was negatively correlated with the absorption of these minerals, although the effects varied. CONCLUSION: Non-digestible disaccharides increase calcium, magnesium, and iron absorption in normal and ovariectomized rats; however, the effects depend on the disaccharide tested, a fact that may be partly associated with the cecal fermentation of these disaccharides.     

The prebiotic effects of biscuits containing partially hydrolysed guar gum and fructo-oligosaccharides--a human volunteer study

Prebiotics are non-digestible food ingredients that target selected groups of the human colonic microflora, thus having the ability to alter the composition towards a more 'beneficial' community, i.e. selectively increasing populations of bifidobacteria and/or lactobacilli. In the present study the prebiotic potential of partially hydrolysed guar gum (PHGG) and fructo-oligosaccharides (FOS) in a biscuit was assessed in human volunteers. Fluorescent in situ hybridization using oligonucleotide probes targeting Bacteroides spp., Bifidobacterium spp., Clostridium spp. and Lactobacillus-Enterococcus spp. were used for the bacteriology and total bacteria were enumerated using the fluorescent stain 4',6-diamidino-2-phenylindole. Thirty-one volunteers consumed daily either three experimental biscuits (providing a total (g/d) of 6.6 FOS and 3.4 PHGG) or three placebo biscuits for two 21-d crossover periods. Bifidobacteria significantly increased in number on ingestion of the experimental biscuits compared with pre-treatment and placebo population levels. Bifidobacterial numbers returned to pretreatment levels within 7 d of the cessation of intake of experimental biscuits. A correlation was observed between the initial faecal bifidobacterial numbers and the magnitude of bifidogenesis, with volunteers who possessed low initial population levels of bifidobacteria experiencing the greatest increase in bifidogenesis. No changes were observed in the other bacterial groups monitored during the trial. Thus, the prebiotic nature of FOS and PHGG was maintained in a final food product as evidenced from the selective increase in bifidobacterial numbers.