Search

The gut microbiota has been indicated to play a crucial role in health and disease. Apart from changes in composition between healthy individuals and those with a disease or disorder, it has become clear that also microbial activity is important for health. For instance, butyrate has been proven to be beneficial for health, because, amongst others, it is a substrate for the colonocytes, and modulates the host’s immune system and metabolism. Here, we studied the effect of a blend of three mushrooms (Ganoderma lucidum GL AM P-38, Grifola frondosa GF AM P36 and Pleurotus ostreatus PO AM-GP37)) on gut microbiota composition and activity in a validated, dynamic, computer-controlled in vitro model of the colon (TIM-2). Predigested mushroom blend at three doses (0.5, 1.0 and 1.5 g/day of ingested mushroom blend) was fed to a pooled microbiota of healthy adults for 72 h, and samples were taken every day for microbiota composition (sequencing of amplicons of the V3-V4 region of the 16S rRNA gene) and activity (short-chain fatty acid (SCFA) production). The butyrate producing genera Lachnospiraceae UCG-004, Lachnoclostridium, Ruminococcaceae UCG-002 and Ruminococcaceae NK4A214-group are all dose-dependently increased when the mushroom blend was fed. Entirely in line with the increase of these butyrate-producers, the cumulative amount of butyrate also dose-dependently increased, to roughly twice the amount compared to the control (medium without mushroom blend) on the high-dose mushroom blend. Butyrate proportionally made up 53.1% of the total SCFA upon feeding the high-dose mushroom blend, compared to 27% on the control medium. In conclusion, the (polysaccharides in the) mushroom blend led to substantial increase in butyrate by the gut microbiota. These results warrant future mechanistic research on the mushroom blend, as butyrate is considered to be one of the microbial metabolites that contributes to health, by increasing barrier function and modulating inflammation.

Sizeable scientific evidence indicates the health benefits related to phenolic compounds and dietary fiber. Various phenolic compounds-rich foods or ingredients are also rich in dietary fiber, and these two health components may interrelate via noncovalent (reversible) and covalent (mostly irreversible) interactions. Notwithstanding, these interactions are responsible for the carrier effect ascribed to fiber toward the digestive system and can modulate the bioaccessibility of phenolics, thus shaping health-promoting effects in vivo. On this basis, the present review focuses on the nature, occurrence, and implications of the interactions between phenolics and food components. Covalent and noncovalent interactions are presented, their occurrence discussed, and the effect of food processing introduced. Once reaching the large intestine, fiber-bound phenolics undergo an intense transformation by the microbial community therein, encompassing reactions such as deglycosylation, dehydroxylation, alpha- and beta-oxidation, dehydrogenation, demethylation, decarboxylation, C-ring fission, and cleavage to lower molecular weight phenolics. Comparatively less information is still available on the consequences on gut microbiota. So far, the very most of the information on the ability of bound phenolics to modulate gut microbiota relates to in vitro models and single strains in culture medium. Despite offering promising information, such models provide limited information about the effect on gut microbes, and future research is deemed in this field. Fundacao para a Ciencia e Tecnologia (FCT)Portuguese Foundation for Science and TechnologyEuropean Commission [PTDC/SAU-NUT/30448/2017, SFRH/BPD/85293/2012] This work was partially supported by PTDC/SAU-NUT/30448/2017 with funding from FundacAo para a Ciencia e Tecnologia (FCT) through national funds. Perez-Gregorio acknowledges her FCT research contract (SFRH/BPD/85293/2012) and LAQV-REQUIMTE as the host institution.

Infusions of the short-chain fatty acid (SCFA) acetate in the distal colon improved metabolic parameters in men. Here, we hypothesized that combining rapidly and slowly fermentable fibers will enhance distal colonic acetate production and improve metabolic health. In vitro cultivation studies in a validated model of the colon were used to identify fiber mixtures that yielded high distal colonic acetate production. Subsequently, in two randomized crossover studies, lean and prediabetic overweight/obese men were included. In one study, participants received supplements of either long-chain inulin+resistant starch (INU+RS), INU or maltodextrin (PLA) the day prior to a clinical investigation day (CID). The second trial studied beta glucan+RS (BG+RS) versus BG and PLA. During each CID, breath hydrogen, indirect calorimetry, plasma metabolites/hormones were assessed during fasting and postprandial conditions. Additionally, fecal microbiota composition and SCFA were determined. In prediabetic men, INU+RS increased plasma acetate compared to INU or PLA (P < .05), but did not affect metabolic parameters. In lean men, INU+RS increased breath hydrogen and fasting plasma butyrate, which was accompanied by increased energy expenditure, carbohydrate oxidation and PYY and decreased postprandial glucose concentrations (all P < .05) compared to PLA. BG+RS increased plasma butyrate compared to PLA (P < .05) in prediabetic individuals, but did not affect other fermentation/metabolic markers in both phenotypes. Fiber-induced shifts in fecal microbiota were individual-specific and more pronounced with INU+RS versus BG+RS. Administration of INU+RS (not BG+RS) the day prior to investigation improved metabolic parameters in lean but not in prediabetic individuals, demonstrating that effects were phenotype- and fiber-specific. Further research should study whether longer-term supplementation periods are required to elicit beneficial metabolic health in prediabetic individuals. Trial registration numbers: Clinical trial No. NCT03711383 (Inulin study) and Clinical trial No. NCT03714646 (Beta glucan study).

The effect of a Citrus Fruit Extract high in the polyphenols hesperidin and naringin (CFE) on modulation of the composition and activity of the gut microbiota was tested in a validated, dynamic in vitro model of the colon (TIM-2). CFE was provided at two doses (250 and 350 mg/day) for 3 days. CFE led to a dose-dependent increase in Roseburia, Eubacterium ramulus, and Bacteroides eggerthii. There was a shift in production of short-chain fatty acids, where acetate production increased on CFE, while butyrate decreased. In overweight and obesity, acetate has been shown to increase fat oxidation when produced in the distal gut, and stimulate secretion of appetite-suppressive neuropeptides. Thus, the data in the in vitro model point towards mechanisms underlying the effects of the polyphenols in CFE with respect to modulation of the gut microbiota, both in composition and activity. These results should be confirmed in a clinical trial.

The aim of the study was to investigate the potential prebiotic effects of food-by-products (cassava bagasse (n=3), orange bagasse (n=2) and passion fruit peel (n=3)) using an in vitro model simulating the proximal colon, and to assess possible differences in fermentation when using faecal microbiota from lean or obese people. Fermentation of the by-products was compared to a control medium and the prebiotic inulin. The effects of the by-products on the dynamics of the gut microbiota differed according to the type of microbiota, as well as the type of by-product used. Principal Coordinate Analysis of the microbiota showed evidence of a clear separate clustering of lean and obese microbiota before the addition of substrates, which disappeared after fermentation, and instead, distinct clusters due to primary carbohydrate composition of the by-products (starch, fructan and pectin) were present. This is evidence that the substrates drove the obese microbiota to a healthier profile, more similar to that of the lean microbiota. Cassava bagasses enriched the beneficial genus Bifidobacterium in the obese microbiota. The production of total SCFA by cassava bagasses by the obese microbiota was higher than for control medium and inulin. Orange bagasses stimulated the growth of the butyrate-producing genus Coprococcus. Passion fruit peels were poorly fermented and generated negligible amounts of intermediate metabolites, indicating slow fermentation. Nevertheless, passion fruit peel fermentation resulted in a microbiota with the highest diversity and evenness, a positive trait regarding host health. In conclusion, the use of food-by-products could be an important step to tackle obesity and decrease the waste of valuable food material and consequently environmental pollution. They are an inexpensive and non-invasive way to be used as a dietary intervention to improve health, as they were shown here to drive the composition of the obese microbiota to a healthier profile.

Diet-related modulation of gut microbiota and its metabolic activity represents an intriguing research context, particularly in the case of disorders related to imbalances in gut microbial communities. We here explored the effects of Bacillus coagulans GBI-30, 6086 (BC30), β-glucans, and innovative whole-grain pastas, with or without these functional ingredients, on gut microbiota from three groups of children, presenting different susceptibility to type 1 diabetes, by using the well-controlled TNO in vitro model of the proximal colon (TIM-2). Short- and branched-chain fatty acids production and microbiota composition were assessed by means of gas chromatography and 16S rRNA gene profiling, respectively. In most cases, in vitro dietary interventions caused microbiota-dependent modulations as a result of intergroup variability, but also specific changes in microbial groups were shared between the three microbiotas, highlighting specific diet-microbial taxa connections.

BACKGROUND & AIMS: Biomarker assays could increase the accuracy of noninvasive detection of colorectal cancer (CRC); fecal immunochemical tests (FITs) are estimated to miss 27%-47% of CRCs and 70%-80% of advanced adenomas per round of screening. We investigated the conditions under which biomarker screens would be cost-effective compared with FIT screens of average-risk individuals. METHODS: We used the MISCAN-Colon microsimulation model to estimate the effects of various CRC screening test characteristics on life-years gained (LYG) and; age-specific all-cause mortality was based on the 2010 Dutch life tables. Simulated CRC incidence rate and CRC stage distribution were calibrated to observed data in The Netherlands from 1999 through 2003 (before opportunities for screening). Survival rates after diagnosis of CRC at an age younger than 75 years were based on CRC relative survival data from 1985 through 2004; survival for individuals diagnosed at an age of 75 years or older was adjusted to fit the observed age-increasing mortality/incidence ratio. We modeled FIT along with hypothetical biomarker tests with different test performance levels. For each biomarker test we calculated the maximum unit cost for the test to be cost-effective compared with FIT, assuming a willingness-to-pay threshold of (sic)50,000 ($ 56,000) per LYG. RESULTS: Biennial FIT screening of subjects 55-75 years old provided 84.9 LYG at a cost of (sic)122,000 ($ 137,000) per 1000 participants. Considering a unit cost of (sic)7 ($ 8) for FIT (including kit and analysis only, excluding organizational costs), a biomarker test that detects CRC with higher levels of specificity and sensitivity (100%) and advanced adenomas at a proportionally higher level of sensitivity (53%) should never exceed a cost of (sic)51 ($ 57). The threshold cost could increase to more than (sic)200 ($ 224) for high-performing biomarker tests in cases of limited colonoscopy capacity or higher uptake of this test. CONCLUSIONS: By using the MISCAN-Colon microsimulation model to estimate effects of CRC screening tests, we foundthat for a biomarker test with increased overall performance to be cost-effective, it should not exceed 7-fold the unit cost of FIT. This maximum would increase substantially if colonoscopy becomes more expensive or scarce, or if the new test has higher screening uptake. These values could be used to estimate the added value of new biomarkers compared with current FIT screening.

The digestive system of the chicken plays an important role in metabolism, immunity, and chicken health and production performance. The chicken ceca harbor a diverse microbial community and play a crucial role in the microbial fermentation and production of energy-rich short-chain fatty acids (SCFA). For humans, dogs, and piglets in vitro digestive system models have been developed and are used to study the microbiota composition and metabolism after intervention studies. For chickens, most research on the cecal microbiota has been performed in in vivo experiments or in static in vitro models that may not accurately resemble the in vivo situations. This paper introduces an optimized digestive system model that simulates the conditions in the ceca of the chicken, i.e., the Chicken ALIMEntary tRact mOdel-2 (CALIMERO-2). The system is based on the well-validated TNO in vitro model of the colon-2 (TIM-2) and is the first dynamic in vitro digestion model for chickens species. To validate this model, the pH, temperature, and different types of microbial feeding were compared and analyzed, to best mimic the conditions in the chicken ceca. The bacterial composition, as well as the metabolite production at 72 h, showed no significant difference between the different microbial feedings. Moreover, we compared the CALIMERO-2 digestive samples to the original inoculum and found some significant shifts in bacterial composition after the fermentation started. Over time the bacterial diversity increased and became more similar to the original inoculum. We can conclude that CALIMERO-2 is reproducible and can be used as a digestive system model for the chicken ceca, in which the microbial composition and activity can be maintained and shows similar results to the in vivo cecum. CALIMERO-2 can be used to study effects on composition and activity of the chicken cecum microbiota in response to in-feed interventions.

Billions of bacteria inhabit the gastrointestinal tract. Immune-microbial cross talk is responsible for immunological homeostasis, and symbiotic microbial species induce regulatory immunity, which helps to control the inflammation levels. In this study we aimed to identify species within the equine intestinal microbiota with the potential to induce regulatory immunity. These could be future targets for preventing or treating low-grade chronic inflammation occurring as a result of intestinal microbial changes and disruption of the homeostasis. 16S rRNA gene amplicon sequencing was performed on samples of intestinal microbial content from ileum, cecum, and colon of 24 healthy horses obtained from an abattoir. Expression of genes coding for IL-6, IL-10, IL-12, IL-17, 18 s, TNFα, TGFβ, and Foxp3 in the ileum and mesenteric lymph nodes was measured by qPCR. Intestinal microbiota composition was significantly different in the cecum and colon compared to the ileum, which contains large abundances of Proteobacteria. Especially members of the Clostridiales order correlated positively with the regulatory T-cell transcription factor Foxp3 and so did the phylum Verrucomicrobia. We conclude that Clostridiales and Verrucomicrobia have the potential to induce regulatory immunity and are possible targets for intestinal microbial interventions aiming at regulatory immunity improvement.

Alterations in colonic motility are implicated in the pathophysiology of bowel disorders, but high-resolution manometry of human colonic motor function has revealed that our knowledge of normal motor patterns is limited. Furthermore, various terminologies and definitions have been used to describe colonic motor patterns in children, adults and animals. An example is the distinction between the high-amplitude propagating contractions in humans and giant contractions in animals. Harmonized terminology and definitions are required that are applicable to the study of colonic motility performed by basic scientists and clinicians, as well as adult and paediatric gastroenterologists. As clinical studies increasingly require adequate animal models to develop and test new therapies, there is a need for rational use of terminology to describe those motor patterns that are equivalent between animals and humans. This Consensus Statement provides the first harmonized interpretation of commonly used terminology to describe colonic motor function and delineates possible similarities between motor patterns observed in animal models and humans in vitro (ex vivo) and in vivo. The consolidated terminology can be an impetus for new research that will considerably improve our understanding of colonic motor function and will facilitate the development and testing of new therapies for colonic motility disorders., This Consensus Statement provides a conceptual and methodological framework to expand research on colonic motility in experimental animals and humans. The work is intended to facilitate the development of new drugs for common colonic motility disorders and of appropriate diagnostic and therapeutic algorithms for the management of paediatric and adult patients.

Short-chain fatty acids (SCFA), such as sodium butyrate (SB), sodium propionate (SPr), and sodium acetate (SAc), are metabolic end-products of the fermentation of dietary fibers. They are linked with multiple beneficial effects on the general mammalian health, based on the sophisticated interplay with the host immune response. Equine herpesvirus 1 (EHV1) is a major pathogen, which primarily replicates in the respiratory epithelium, and disseminates through the body via a cell-associated viremia in leukocytes, even in the presence of neutralizing antibodies. Infected monocytic CD172a(+) cells and T-lymphocytes transmit EHV1 to the endothelium of the endometrium or central nervous system (CNS), causing reproductive or neurological disorders. Here, we questioned whether SCFA have a potential role in shaping the pathogenesis of EHV1 during the primary replication in the URT, during the cell-associated viremia, or at the level of the endothelium of the pregnant uterus and/or CNS. First, we demonstrated the expression of SCFA receptors, FFA2 and FFA3, within the epithelium of the equine respiratory tract, at the cell surface of immune cells, and equine endothelium. Subsequently, EHV1 replication was evaluated in the URT, in the presence or absence of SB, SPr, or SAc. In general, we demonstrated that SCFA do not affect the number of viral plaques or virus titer upon primary viral replication. Only SB and SPr were able to reduce the plaque latitudes. Similarly, pretreatment of monocytic CD172a(+) cells and T-lymphocytes with different concentrations of SCFA did not alter the number of infected cells. When endothelial cells were treated with SB, SPr, or SAc, prior to the co-cultivation with EHV1-inoculated mononuclear cells, we observed a reduced number of adherent immune cells to the target endothelium. This was associated with a downregulation of endothelial adhesion molecules ICAM-1 and VCAM-1 in the presence of SCFA, which ultimately lead to a significant reduction of the EHV1 endothelial plaques. These results indicate that physiological concentrations of SCFA may affect the pathogenesis of EHV1, mainly at the target endothelium, in favor of the fitness of the horse. Our findings may have significant implications to develop innovative therapies, to prevent the devastating clinical outcome of EHV1 infections.

In vitro gas production studies are routinely used to assess the metabolic capacity of intestinal microbiota to ferment dietary fibre sources. The faecal inocula used during the in vitro gas production procedure are most often obtained from animals adapted to a certain diet. The present study was designed to assess whether 19 days of adaptation to a diet are sufficient for faecal inocula of pigs to reach a stable microbial composition and activity as determined by in vitro gas production. Eighteen multiparous sows were allotted to one of two treatments for three weeks: a diet high in fibre (H) or a diet low in fibre (L). After this 3-week period, the H group was transferred to the low fibre diet (HL-treatment) while the L group was transferred to the diet high in fibre (LH-treatment). Faecal samples were collected from each sow at 1, 4, 7, 10, 13, 16 and 19 days after the diet change and prepared as inoculum used for incubation with three contrasting fermentable substrates: oligofructose, soya pectin and cellulose. In addition, inocula were characterised using a phylogenetic microarray targeting the pig gastrointestinal tract microbiota. Time after diet change had an effect (P

International audience; Faeces could be used for evaluating the balance of the equine hindgut microbial ecosystem, which would offer a practical method for assessing gut health and how this relates to disease. However, previous studies concluded that faeces microbial ecosystem was not representative of the proximal hindgut (caecum and ventral colon). This study aimed to evaluate if variations of the faecal microbial ecosystem were similar to those observed in the proximal hindgut. Six horses, fistulated in the caecum and right ventral (RV) colon, were subjected to a gradual change of diet, from a 100% hay (high fibre) diet (2.2 DM kg/day per 100 kg BW) to a 57% hay + 43% barley (high starch) diet (0.8 DM kg/day per 100 kg BW hay and 0.6 DM kg/day per 100 kg BW barley). The two diets were iso-energetic and fed over a 3-week trial period. Samples of digesta from the caecum, RV colon and faeces were collected two times on the 10th and 20th day of the trial, for each diet to assess the microbial ecosystem parameters by both classical culture technics and biochemical methods. The variations observed in the caecal and colonic bacterial composition (increase in total anaerobic, amylolytic and lactate-utilizing and decrease in cellulolytic bacteria concentrations) and microbial activity (changes in volatile fatty acids concentrations and increase in lactate concentrations) demonstrated that the hay+barley diet caused changes in the hindgut microbial ecosystem. Similar variations were observed in the faecal microbial ecosystem. Feeding the hay+barley diet resulted in higher concentrations of faecal lipopolysaccharides. The functional bacterial group concentrations (cellulolytics, amylolytics and lactate utilizers) were significantly correlated between caecum and faeces and between colon and faeces. From analyses of the metabolites produced from microbial activity, only valerate concentration in the caecum and the proportion of propionate were significantly correlated with the same parameters in the faeces. Results of the principal component analysis performed between all the caecal/faecal and colonic/faecal parameters revealed that the total anaerobic and cellulolytic bacteria concentrations, as well as valerate, L-lactate and lipopolysaccharide concentrations were strongly correlated with several microbial parameters in the caecum (P < 0.027; r >|0.45|) and in the colon (P < 0.013; r > |0.50|). This demonstrated that faecal samples and their bacterial analyses could be used to represent caecum and RV colon hindgut microbial ecosystem in terms of variations during a change from a high-fibre to a high-starch diet, and thus could be markers of particular interest to diagnostic proximal hindgut microbial disturbances.

Scope: We aimed to investigate the effects of three different soluble pectins on the digestion of other consumed carbohydrates, and the consequent alterations of microbiota composition and SCFA levels in the intestine of pigs.Methods and results: Piglets were fed a low-methyl esterified pectin enriched diet (LMP), a high-methyl esterified pectin enriched diet (HMP), a hydrothermal treated soybean meal enriched diet (aSBM) or a control diet (CONT). LMP significantly decreased the ileal digestibility of starch resulting in more starch fermentation in the proximal colon. In the ileum, low-methyl esterified pectin present was more efficiently fermented by the microbiota than high-methyl esterified pectin present which was mainly fermented by the microbiota in the proximal colon. Treated soybean meal was mainly fermented in the proximal colon and shifted the fermentation of cereal dietary fiber to more distal parts, resulting in high SCFA levels in the mid colon. LMP, HMP, and aSBM decreased the relative abundance of the genus Lactobacillus and increased that of Prevotella in the colon.Conclusion: The LMP, HMP, and aSBM, differently affected the digestion processes compared to the control diet and shaped the colonic microbiota from a Lactobacillus-dominating flora to a Prevotella-dominating community, with potential health-promoting effects.

The aim of this research was to study the in vitro fermentation of chicory root pulp (CRP) and ensiled CRP (ECRP) using human fecal inoculum. Analysis of carbohydrate levels in fermentation digests showed that 51% of all CRP carbohydrates were utilized after 24 h of fermentation. For ECRP, having the same cell wall polysaccharide composition as CRP, but with solubilization of 4 times more of CRP pectin due to ensiling, the fermentation was quicker than with CRP as 11% more carbohydrates were utilized within the first 12 h. The level of fiber utilization for ECRP after 24 h was increased by 8% compared to CRP. This effect on fiber utilization from ECRP seemed to arise from (i) increased levels of soluble pectin fibers (arabinan, homogalacturonan, and galactan) and (ii) ahypothesized more open structure of the remaining cell walls in ECRP, which was more accessible to degradation than the CRP cell wall network. KEYWORDS: chicory root pulp, ensiled chicory root pulp, insoluble

Commonly used feed processing technologies are not sufficient to affect recalcitrant non-starch polysaccharides (NSP) such as arabinoxylans present in maize distillers dried grain with solubles (DDGS). Instead, hydrothermal treatments combined with acid catalysts might be more effective to modify these NSP. The objective of this experiment was to investigate the effects of hydrothermal maleic acid treatment (acid-extrusion) on the degradability of maize DDGS in growing pigs. It was hypothesized that acid-extrusion modifies DDGS cell wall architecture and, thereby, increases fermentability of NSP. Two diets, containing either 40% (wt/wt) unprocessed or acid-extruded DDGS, were restrictedly fed to groups of gilts (n = 11, with 4 pigs per group; initial mean BW: 20.8 ± 0.2 kg) for 18 d and performance and digestibility were analyzed. Acid-extrusion tended to decrease apparent ileal digestibility (AID) of CP (~3 percentage units, P = 0.063) and starch (~1 percentage unit, P = 0.096). Apparent digestibility of CP and starch measured at the mid colon (2 percentage units, P = 0.030 for CP and 0.3 percentage units, P

Objective To estimate benefits and harms of different colorectal cancer screening strategies, stratified by (baseline) 15-year colorectal cancer risk. Design Microsimulation modelling study using MIcrosimulation SCreening ANalysis-Colon (MISCAN-Colon). Setting A parallel guideline committee ( BMJ Rapid Recommendations) defined the time frame and screening interventions, including selection of outcome measures. Population Norwegian men and women aged 50-79 years with varying 15-year colorectal cancer risk (1-7%). Comparisons Four screening strategies were compared with no screening: biennial or annual faecal immunochemical test (FIT) or single sigmoidoscopy or colonoscopy at 100% adherence. Main outcome measures Colorectal cancer mortality and incidence, burdens, and harms over 15 years of follow-up. The certainty of the evidence was assessed using the GRADE approach. Results Over 15 years of follow-up, screening individuals aged 50-79 at 3% risk of colorectal cancer with annual FIT or single colonoscopy reduced colorectal cancer mortality by 6 per 1000 individuals. Single sigmoidoscopy and biennial FIT reduced it by 5 per 1000 individuals. Colonoscopy, sigmoidoscopy, and annual FIT reduced colorectal cancer incidence by 10, 8, and 4 per 1000 individuals, respectively. The estimated incidence reduction for biennial FIT was 1 per 1000 individuals. Serious harms were estimated to be between 3 per 1000 (biennial FIT) and 5 per 1000 individuals (colonoscopy); harms increased with older age. The absolute benefits of screening increased with increasing colorectal cancer risk, while harms were less affected by baseline risk. Results were sensitive to the setting defined by the guideline panel. Because of uncertainty associated with modelling assumptions, we applied a GRADE rating of low certainty evidence to all estimates. Conclusions Over a 15 year period, all screening strategies may reduce colorectal cancer mortality to a similar extent. Colonoscopy and sigmoidoscopy may also reduce colorectal cancer incidence, while FIT shows a smaller incidence reduction. Harms are rare and of similar magnitude for all screening strategies.

Background: High-amylose-maize resistant starch type 2 (HAMRS2) is a fermentable dietary fiber known to alter the gut milieu, including the gut microbiota, which may explain the reported effects of resistant starch to ameliorate obesity associated metabolic dysfunction.Objective: Our working hypothesis was that HAMRS2-induced microbiome changes alter gut-derived signals (i.e., xenometabolites) reaching the liver via the portal circulation, in turn altering liver metabolism by regulating gene expression and other pathways.Methods: We used a multi-omics systems biology approach to characterize HAMRS2-driven shifts to the cecal microbiome, liver metabolome, and transcriptome, identifying correlates between microbial changes and liver metabolites under obesogenic conditions that, to our knowledge, have not previously been recognized. Five-week-old male C57BL/6J mice were fed an energy-dense 45% lard-based-fat diet for 10 wk supplemented with either 20% HAMRS2 by weight (n = 14) or rapidly digestible starch (control diet; n = 15).Results: Despite no differences in food intake, body weight, glucose tolerance, fasting plasma insulin, or liver triglycerides, the HAMRS2 mice showed a 15-58% reduction in all measured liver amino acids, except for Gln, compared with control mice. These metabolites were equivalent in the plasma of HAMRS2 mice compared with controls, and transcripts encoding key amino acid transporters were not different in the small intestine or liver, suggesting that HAMRS2 effects were not simply due to lower hepatocyte exposure to systemic amino acids. Instead, alterations in gut microbial metabolism could have affected host nitrogen and amino acid homeostasis: HAMRS2 mice showed a 62% increase (P Conclusion: Together, these differences indicate that HAMRS2 dramatically alters hepatic metabolism and gene expression concurrent with shifts in specific gut bacteria in C57BL/6J mice.

Background An aberrant metabolic activity or a compositional alteration of the gut microbiota has been proposed as a factor that makes us more prone to disease. Therefore, we explored the effect of two dietary fibers (arabinogalactan and inulin) on the microbiota from lean and obese subjects during 72 h in vitro fermentation experiments using the validated TNO dynamic in vitro model of the proximal colon: TIM-2. Metabolically, arabinogalactan fermentation showed a higher production of propionate when compared to n-butyrate in the obese microbiota fermentations. In general, lean microbiota produced more n-butyrate from the fermentation of both substrates when compared to the obese microbiota. Furthermore, the obese microbiota extracted more energy from the fermentation of both fibers. Results Compositionally, bacteria belonging to Gemmiger, Dorea, Roseburia, Alistipes, Lactobacillus and Bifidobacterium genera were found to be highly abundant or stimulated by the prebiotics in the lean microbiota suggesting a potential role in leanness. Furthermore, a significant correlation between known butyrogenic strains including B. adolescentis,an unclassified Bifidobacterium and F. prausnitzii with this metabolite in the fermentation of inulin in both microbiotas was found. Conclusions Although supplementary in vivo studies are needed, the current study provides more evidence for the consumption of specific ingredients with the aim of modulating the gut microbiota in the context of obesity. © 2016 Aguirre et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

The natural diet of felids contains highly digestible animal tissues but also fractions resistant to small intestinal digestion, which enter the large intestine where they may be fermented by the resident microbial population. Little information exists on the microbial degradability of animal tissues in the large intestine of felids consuming a natural diet. This study aimed to rank animal substrates in their microbial degradability by means of an in vitro study using captive cheetahs fed a strict carnivorous diet as fecal donors. Fresh cheetah fecal samples were collected, pooled, and incubated with various raw animal substrates (chicken cartilage, collagen, glucosamine-chondroitin, glucosamine, rabbit bone, rabbit hair, and rabbit skin; 4 replicates per substrate) for cumulative gas production measurement in a batch culture technique. Negative (cellulose) and positive (casein and fructo-oligosaccharides; FOS) controls were incorporated in the study. Additionally, after 72 h of incubation, short-chain fatty acids (SCFA), including branched-chain fatty acids (BCFA), and ammonia concentrations were determined for each substrate. Glucosamine and glucosamine-chondroitin yielded the greatest OM cumulative gas volume (OMCV) among animal substrates (P

This study introduces a stable-isotope metabolic approach employing [U-(13)C]glucose that, as a novelty, allows selective profiling of the human intestinal microbial metabolic products of carbohydrate food components, as well as the measurement of the kinetics of their formation pathways, in a single experiment. A well-established, validated in vitro model of human intestinal fermentation was inoculated with standardized gastrointestinal microbiota from volunteers. After culture stabilization, [U-(13)C]glucose was added as an isotopically labeled metabolic precursor. System lumen and dialysate samples were taken at regular intervals. Metabolite concentrations and isotopic labeling were determined by NMR, GC, and enzymatic methods. The main microbial metabolites were lactate, acetate, butyrate, formate, ethanol, and glycerol. They together accounted for a (13)C recovery rate as high as 91.2%. Using an NMR chemical shift prediction approach, several minor products that showed (13)C incorporation were identified as organic acids, amino acids, and various alcohols. Using computer modeling of the (12)C contents and (13)C labeling kinetics, the metabolic fluxes in the gut microbial pathways for synthesis of lactate, formate, acetate, and butyrate were determined separately for glucose and unlabeled background substrates. This novel approach enables the study of the modulation of human intestinal function by single nutrients, providing a new rational basis for achieving control of the short-chain fatty acids profile by manipulating substrate and microbiota composition in a purposeful manner.

BACKGROUND: In organic farming, roughage has to be added to pigs' daily rations for animal welfare reasons. Since little is known about how roughage affects pigs, seven carbohydrate-rich feedstuffs (corn silage, ryegrass, turnip leaf, turnip tuber, Jerusalem artichoke and two types of clover-ryegrass silage) were tested using two different in vitro methods - total tract digestibility and the gas production technique - to determine apparent digestibility by observing organic matter loss. The aim was to determine whether the results of different methods and treatments lead to similar results or rankings. RESULTS: The in vitro enzymatic incubation method showed 3-20% lower fermentabilities than the in vitro fermentation process. The pretreatment of substrates with enzymes before fermentation led to similar fermentabilities but different gas production kinetics compared to the fermentation of untreated substrates. Using two different inocula generated no significant differences. When the feedstuffs were ranked by their fermentability, the rank order was relatively similar for both methods. CONCLUSIONS: Different methods and pretreatments lead to different fermentabilities while a ranking of the results gives a similar order. Both methods seem to be appropriate tools for assessing feedstuffs. In order to determine which feedstuff is best utilised by the animal, further investigations of both digestibility and gas production kinetics are required

An experiment was conducted to examine changes in VFA and ammonia concentrations at different time points using 4 fermentable carbohydrate-rich feed ingredients as substrates and feces of unweaned piglets as inoculum. Fecal inoculum was collected, pooled, and mixed from 9 specially raised (no creep feed or antibiotics) crossbred piglets at 3 wk of age. Inulin, lactulose, molasses-free sugar beet pulp, and wheat starch were used as substrates and were fermented in vitro for 72 h (3 replicates per substrate). Cumulative gas production was measured as an indicator of the kinetics of fermentation. In addition, 3 bottles of substrate per time point with similar contents (amounts of substrate, inoculum, and media) were incubated but were allowed to release their gas throughout incubation. For these latter bottles, fermentation fluid was sampled at incubation time points including every hour between 1 and 24 h and at 48 h, and fermentation end products (VFA, lactate, and ammonia) and OM disappearance were measured. Dry matter and ash were analyzed from the postfermentative samples. The pH of the contents from these bottles was also recorded. The correlation in time between fermentation end products and cumulative gas produced was determined. The results showed that the prolongation of fermentation to 72 h, especially in the case of fast-fermenting inulin and lactulose, may lead to a different end product profile (P < 0.001) compared with the profile observed at the time at which most of the substrate has disappeared. Therefore, we concluded that the fermentation product profile at the end of in vitro fermentation at a specific time point cannot be used to compare fermentability of carbohydrate sources with different fermentation kinetics in terms of gas production.

An experiment was conducted to examine changes in VFA and ammonia concentrations at different time points using 4 fermentable carbohydrate-rich feed ingredients as substrates and feces of unweaned piglets as inoculum. Fecal inoculum was collected, pooled, and mixed from 9 specially raised (no creep feed or antibiotics) crossbred piglets at 3 wk of age. Inulin, lactulose, molasses-free sugar beet pulp, and wheat starch were used as substrates and were fermented in vitro for 72 h (3 replicates per substrate). Cumulative gas production was measured as an indicator of the kinetics of fermentation. In addition, 3 bottles of substrate per time point with similar contents (amounts of substrate, inoculum, and media) were incubated but were allowed to release their gas throughout incubation. For these latter bottles, fermentation fluid was sampled at incubation time points including every hour between 1 and 24 h and at 48 h, and fermentation end products (VFA, lactate, and ammonia) and OM disappearance were measured. Dry matter and ash were analyzed from the postfermentative samples. The pH of the contents from these bottles was also recorded. The correlation in time between fermentation end products and cumulative gas produced was determined. The results showed that the prolongation of fermentation to 72 h, especially in the case of fast-fermenting inulin and lactulose, may lead to a different end product profile (P

An experiment was conducted to examine differences in the in vitro fermentability of four carbohydrate-rich feed ingredients and two weaning piglet diets with and without these ingredients, using both the ileal contents and the faeces of unweaned piglets as inocula. In the first part of the experiment, cumulative gas production was measured over time, using faecal inocula mixed from nine specially raised crossbred piglets (no creep feed or antibiotics) at 3 weeks of age. Inulin, lactulose, unmolassed sugar beet pulp, wheat starch and the two diets were used as substrates and fermented in vitro for 72 h. Gas production was measured as an indicator of the kinetics of fermentation. Fermentation end-products, including volatile fatty acids and ammonia, and organic matter loss, were also measured. For the fermentations of feed ingredients, samples were also collected for polymerase chain reaction/denaturing gradient gel electrophoresis analyses initially and after the fermentation process, to study changes in the composition of the bacterial community. This procedure was repeated 1 week later, using ileal contents from the same piglets as inoculum. There were significant differences between the inocula, in terms of both overall fermentation characteristics and composition and between the substrates. There was also a significant interaction between inocula and substrates, which suggests that there were potentially important differences in the microbial activity occurring in these two areas of the gastrointestinal tract (GIT). For the two diets, one with and one without addition of these fermentable ingredients, there were significant differences in terms of the kinetics, but less so in terms of the end-products of fermentation. It was concluded that inocula from both the small and large intestine should be used to obtain a more accurate assessment of potential feed ingredients which will stimulate fermentation in the piglet GIT. Copyright © 2005 Society of Chemical Industry

Recently, it has become apparent that GIT fermentation is not only of interest for ruminant animals, but also for monogastrics. While it is now widely accepted that the fermentation process and its resultant end-products can have important influences on animal health, little is known about the microbiological and immunological processes involved. In terms of animal health, most interest at the moment is focussed on those moments in animals' lives when they are faced with sudden changes resulting in stress. The period of weaning in piglets is a typical example of this. The most easily accomplished and appropriate way to influence GIT fermentation processes is that of dietary intervention. This is reflected by the widespread interest in so-called pre- and pro-biotics. Given the complexities of the interactions occurring in the animal itself, it is hardly surprising that in vitro techniques are being widely used: firstly to examine potential substrates for their fermentability and possible inclusion in diets, and secondly, to assess changes in the microbial populations in response to these substrates. This paper will review the techniques currently in use for these two aspects of monogastric fermentation, and provide examples of their use.

Background: Hereditary non-polyposis colorectal cancer (HNPCC) is thought to arise from adenomas. HNPCC mostly occurs in the proximal colon. We investigated whether this proximal preponderance is due to a proximal preponderance of adenomas or (also) differences in transformation rates from adenomas to cancer between the distal and proximal colon. Methods: A total of 100 HNPCC adenomas were evaluated and compared with 152 sporadic adenomas for location, size, and dysplasia. Twenty five adenomas from patients with a known mismatch repair (MMR) gene mutation were stained for expression of MLH1 and MSH2. Results: HNPCC adenomas were more often located proximally (50% v 26%; p=0.018) and were smaller in comparison with sporadic adenomas. They were similarly dysplastic. However, all proximal HNPCC adenomas ≥5 mm were highly dysplastic compared with 17% of the larger proximal sporadic polyps (p

Background: Hereditary non-polyposis colorectal cancer (HNPCC) is thought to arise from adenomas. HNPCC mostly occurs in the proximal colon. We investigated whether this proximal preponderance is due to a proximal preponderance of adenomas or (also) differences in transformation rates from adenomas to cancer between the distal and proximal colon.Methods: A total of 100 HNPCC adenomas were evaluated and compared with 152 sporadic authors' affiliations adenomas for location, size, and dysplasia. Twenty five adenomas from patients with a known mismatch repair (MMR) gene mutation were stained for expression of MLH1 and MSH2.Results: HNPCC adenomas were more often located proximally (50% v 26%; p=0.018) and were smaller in comparison with sporadic adenomas. They were similarly dysplastic. However, all proximal HNPCC adenomas greater than or equal to5 mm were highly dysplastic compared with 17% of the larger proximal sporadic polyps (pConclusion: HNPCC adenomas are located mainly in the proximal colon. The progression to high grade dysplasia is more common in proximal than distal HNPCC adenomas, indicating a faster transformation rate from early adenoma to cancer in the proximal colon. MMR gene malfunction probably does not initiate adenoma development but is present at a very early stage of tumorigenesis and heralds the development of high grade dysplasia.

The aim of this study was to investigate the effect of galacto-oligosaccharides, lactulose, apple fiber and sugar beet pectin on the composition and activity of human colonic microbiota of lean and obese healthy subjects using an in vitro model of the proximal colon: TIM-2. Substrate fermentation was assessed by measuring the production of short-chain and branched-chain fatty acids, lactate and ammonia and by studying the composition of the bacterial communities over time. The results suggest that energy harvest (in terms of metabolites) of lean and obese microbiotas is different and may depend on the fermentable substrate. For galactooligosaccharides and lactulose, the cumulative amount of short-chain fatty acids plus lactate produced in TIM-2 was lower in the fermentation experiments with the lean microbiota (123 and 155 mmol, respectively) compared to the obese (162 and 173 mmol, respectively). This was reversed for the pectin and the fiber. The absolute amount produced of short-chain fatty acids including lactate was higher after 72 h in the fermentation experiments with apple fiber-L (108 mmol) than with apple fiber-O (92 mmol). Sugar beet-L was also higher (130 mmol) compared to sugar beet-O (103 mmol). Galacto-oligosaccharides and lactulose boosted the balance of health-promoting over toxic metabolites produced by the microbiota from obese subjects. Firmicutes were more predominant in the inoculum prepared from feces of obese subjects compared to lean subjects. The average abundance at time zero was 92% and 74%, respectively. On the other hand, Bacteroidetes were more dominant in the microbiota prepared with homogenates from lean subjects with an average abundance of 22% compared with the microbiota prepared with homogenates from obese subjects (3.6%). This study brings evidence that different fermentable carbohydrates are fermented differently by lean and obese microbiotas, which contributes to the understanding of the role of diet and the microbiota in tackling obesity. Copyright

The in vitro fermentation characteristics of different classes of sugar beet pectic oligosaccharides (SBPOS) were studied using human and pig fecal inocula. The SBPOS consisted mainly of partially acetylated rhamnogalacturonan oligosaccharides and partially methyl-esterified/acetylated homogalacturonan oligosaccharides. Some SBPOS contained an unsaturated galacturonic acid residue at their non-reducing end. It was shown that SBPOS could be completely fermented by human and pig fecal microbiota, thereby producing butyrate yet mainly acetate and propionate as metabolites. The degradation of SBPOS by pig fecal microbiota was different and much slower compared to human fecal microbiota. In general, rhamnogalacturonan oligosaccharides were degraded slower than homogalacturonan oligosaccharides. Acetylation of rhamnogalacturonan oligosaccharides lowered the degradation rate by pig fecal microbiota but not by human fecal microbiota. No classic bifidogenic effect was shown for SBPOS using human fecal inoculum. However, several other potentially interesting modifications in the microbiota composition that can be associated with host health were observed, which are discussed. © 2014 American Chemical Society.

Commonly used feed processing technologies are not sufficient to affect recalcitrant non-starch polysaccharides (NSP) such as arabinoxylans present in maize distillers dried grain with solubles (DDGS). Instead, hydrothermal treatments combined with acid catalysts might be more effective to modify these NSP. The objective of this experiment was to investigate the effects of hydrothermal maleic acid treatment (acid-extrusion) on the degradability of maize DDGS in growing pigs. It was hypothesized that acid-extrusion modifies DDGS cell wall architecture and, thereby, increases fermentability of NSP. Two diets, containing either 40% (wt/wt) unprocessed or acid-extruded DDGS, were restrictedly fed to groups of gilts (n = 11, with 4 pigs per group; initial mean BW: 20.8 ± 0.2 kg) for 18 d and performance and digestibility were analyzed. Acid-extrusion tended to decrease apparent ileal digestibility (AID) of CP (~3 percentage units, P = 0.063) and starch (~1 percentage unit, P = 0.096). Apparent digestibility of CP and starch measured at the mid colon (2 percentage units, P = 0.030 for CP and 0.3 percentage units, P

Background: Chronic use of sennoside laxatives often causes pseudomelanosis coli. A recent study suggested that pseudomelanosis coli is associated with an increased colorectal cancer risk. A single high dose of highly purified senna extract increased proliferation rate and reduced crypt length in the sigmoid colon compared to historical controls. Aims: To evaluate in a controlled study the effects of highly purified senna extract on cell proliferation and crypt length in the entire colon and on p53 and bcl-2 expression. Methods: Addition of a senna extract to colonic lavage was studied in 184 consecutive outpatients. From 32 randomised patients, 15 with sennosides (Sen), 17 without (NSen), biopsies were taken. Proliferative activity was studied in 4 areas of the colon, using 5-bromo-2′-deoxyuridine labelling and immunohistochemistry (labelling index, LI). Expression of p53 and bcl-2 in the sigmoid colon was determined immunohistochemically. Results: Crypts were shorter in Sen than in NSen in the transverse and sigmoid colon. LI was higher in Sen than in NSen in the entire colon. No difference in p53 expression was seen. Bcl-2 expression was higher in both groups when crypts were shorter and/or proliferation was increased. Conclusion: Sennosides induce acute massive cell loss probably by apoptosis, causing shorter crypts, and increased cell proliferation and inhibition of apoptosis to restore cellularity. These effects may reflect the mechanism for the suggested cancer-promoting effect of chronic sennoside use.

Background Recent scientific developments have shed more light on the importance of the host-microbe interaction, particularly in the gut. However, the mechanistic study of the host-microbe interplay is complicated by the intrinsic limitations in reaching the different areas of the gastrointestinal tract (GIT) in vivo. In this paper, we present the technical validation of a new device - the Host-Microbiota Interaction (HMI) module - and the evidence that it can be used in combination with a gut dynamic simulator to evaluate the effect of a specific treatment at the level of the luminal microbial community and of the host surface colonization and signaling. Results The HMI module recreates conditions that are physiologically relevant for the GIT: i) a mucosal area to which bacteria can adhere under relevant shear stress (3 dynes cm−2); ii) the bilateral transport of low molecular weight metabolites (4 to 150 kDa) with permeation coefficients ranging from 2.4 × 10−6 to 7.1 × 10−9 cm sec−1; and iii) microaerophilic conditions at the bottom of the growing biofilm (PmO2 = 2.5 × 10−4 cm sec−1). In a long-term study, the host’s cells in the HMI module were still viable after a 48-hour exposure to a complex microbial community. The dominant mucus-associated microbiota differed from the luminal one and its composition was influenced by the treatment with a dried product derived from yeast fermentation. The latter - with known anti-inflammatory properties - induced a decrease of pro-inflammatory IL-8 production between 24 and 48 h. Conclusions The study of the in vivo functionality of adhering bacterial communities in the human GIT and of the localized effect on the host is frequently hindered by the complexity of reaching particular areas of the GIT. The HMI module offers the possibility of co-culturing a gut representative microbial community with enterocyte-like cells up to 48 h and may therefore contribute to the mechanistic understanding of host-microbiome interactions.

A validated in vitro model of the large intestine (TIM-2), set up with human or pig faeces, was used to evaluate the impact of potentially probiotic Lactobacillus amylovorus DSM 16698, administered alone (i), in the presence of prebiotic galactooligosaccharides (GOS) (ii), and co-administered with probiotic Bifidobacterium animalis ssp. lactis Bb-12 (Bb-12) (iii) on GOS degradation, microbial growth (L. amylovorus, lactobacilli, bifidobacteria and total bacteria) and metabolite production. High performance anion exchange chromatography revealed that GOS degradation was more pronounced in TIM-2 inoculated with pig faeces than with human faeces. Denaturing gradient gel electrophoresis profiling of PCR-amplified 16S rRNA genes detected a more complex Lactobacillus spp. community in pig faecal material than in human faecal inoculum. According to 16S rRNA gene-targeted qPCR, GOS stimulated the growth of lactobacilli and bifidobacteria in faecal material from both materials. The cumulative production of short chain fatty acids and ammonia was higher (P < 0.05) for pig than for human faeces. However, lactate accumulation was higher (P < 0.05) in the human model and increased after co-administration with GOS and Bb-12. This study reinforced the notion that differences in microbiota composition between target host organisms need to be considered when animal data are extrapolated to human, as is often done with pre- and probiotic intervention studies. © 2012 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.

The present study intended to evaluate the influence of the collection time of faeces on in vitro fermentation determined with the gas production technique and on the bacteriological counts of the fresh faeces, utilized as inoculum source. Three adult horses, fed hay and concentrates 3 times a day, were used as faeces donors. Fermentation of 5 feedstuffs (barley, oats, alfalfa, meadow hay, and wheat straw), was studied after four collection times: immediately before the morning meal (9:00 h, t0), 2 h after the morning meal (11:00 h, t2), 2 h after the 12:00 h meal (t5) and immediately before the 17:00 h meal (t8). Although there were no differences in pH and bacteriological counts, the proportion in volatile fatty acids, except for acetate, and the extent of fermentation were the highest for t2 (P

The natural diet of felids contains highly digestible animal tissues but also fractions resistant to small intestinal digestion, which enter the large intestine where they may be fermented by the resident microbial population. Little information exists on the microbial degradability of animal tissues in the large intestine of felids consuming a natural diet. This study aimed to rank animal substrates in their microbial degradability by means of an in vitro study using captive cheetahs fed a strict carnivorous diet as fecal donors. Fresh cheetah fecal samples were collected, pooled, and incubated with various raw animal substrates (chicken cartilage, collagen, glucosamine-chondroitin, glucosamine, rabbit bone, rabbit hair, and rabbit skin; 4 replicates per substrate) for cumulative gas production measurement in a batch culture technique. Negative (cellulose) and positive (casein and fructo-oligosaccharides; FOS) controls were incorporated in the study. Additionally, after 72 h of incubation, short-chain fatty acids (SCFA), including branched-chain fatty acids (BCFA), and ammonia concentrations were determined for each substrate. Glucosamine and glucosamine-chondroitin yielded the greatest OM cumulative gas volume (OMCV) among animal substrates (P

This study introduces a stable-isotope metabolic approach employing [U-(13)C]glucose that, as a novelty, allows selective profiling of the human intestinal microbial metabolic products of carbohydrate food components, as well as the measurement of the kinetics of their formation pathways, in a single experiment. A well-established, validated in vitro model of human intestinal fermentation was inoculated with standardized gastrointestinal microbiota from volunteers. After culture stabilization, [U-(13)C]glucose was added as an isotopically labeled metabolic precursor. System lumen and dialysate samples were taken at regular intervals. Metabolite concentrations and isotopic labeling were determined by NMR, GC, and enzymatic methods. The main microbial metabolites were lactate, acetate, butyrate, formate, ethanol, and glycerol. They together accounted for a (13)C recovery rate as high as 91.2%. Using an NMR chemical shift prediction approach, several minor products that showed (13)C incorporation were identified as organic acids, amino acids, and various alcohols. Using computer modeling of the (12)C contents and (13)C labeling kinetics, the metabolic fluxes in the gut microbial pathways for synthesis of lactate, formate, acetate, and butyrate were determined separately for glucose and unlabeled background substrates. This novel approach enables the study of the modulation of human intestinal function by single nutrients, providing a new rational basis for achieving control of the short-chain fatty acids profile by manipulating substrate and microbiota composition in a purposeful manner.

Increased carbohydrate fermentation, compared with protein fermentation, could benefit gut health. In two in vitro experiments, the effect of carbohydrate-rich feed ingredients on fermentation characteristics of ileal chyme from pigs was assessed, using the cumulative gas production technique. Ingredients of the first experiment included gums, inulins, pectins, transgalacto-oligosaccharides, lactose and xylan. In the second experiment, a gum, pectin and transgalacto-oligosaccharides were added at different starting weights, to determine their effects on fermentation characteristics of chyme, in relation to differences in the carbohydrate concentrations. In comparison to fermentation of chyme alone, added carbohydrates led to higher total gas production (p < 0.05), faster maximum rate of gas production (except for xylan) (p < 0.05), and a decreased branched-chain fatty acids to straight chain fatty acids ratio (BCR) (p < 0.05). In the second experiment, for all carbohydrate ingredients, the BCR decreased with increasing starting weights (p < 0.05). If these supplemented dietary carbohydrates were to reach the terminal ileum of the living animal, carbohydrate fermentation in the large intestine could be stimulated, which is known to have beneficial effects on host health.

BACKGROUND: In organic farming, roughage has to be added to pigs' daily rations for animal welfare reasons. Since little is known about how roughage affects pigs, seven carbohydrate-rich feedstuffs (corn silage, ryegrass, turnip leaf, turnip tuber, Jerusalem artichoke and two types of clover-ryegrass silage) were tested using two different in vitro methods - total tract digestibility and the gas production technique - to determine apparent digestibility by observing organic matter loss. The aim was to determine whether the results of different methods and treatments lead to similar results or rankings. RESULTS: The in vitro enzymatic incubation method showed 3-20% lower fermentabilities than the in vitro fermentation process. The pretreatment of substrates with enzymes before fermentation led to similar fermentabilities but different gas production kinetics compared to the fermentation of untreated substrates. Using two different inocula generated no significant differences. When the feedstuffs were ranked by their fermentability, the rank order was relatively similar for both methods. CONCLUSIONS: Different methods and pretreatments lead to different fermentabilities while a ranking of the results gives a similar order. Both methods seem to be appropriate tools for assessing feedstuffs. In order to determine which feedstuff is best utilised by the animal, further investigations of both digestibility and gas production kinetics are required

8 pages, 3 figures, 4 tables.-- PMID: 17766450 [PubMed]., The human intestine harbors both lactate-producing and lactate-utilizing bacteria. Lactate is normally present at, The Rowett Research Institute and Biomathematics and Statistics Scotland are supported by the Scottish Executive Environment and Rural Affairs Department. A. Belenguer received financial support from the Spanish Ministry of Education and Science.

An in vitro experiment was conducted to evaluate the differences in microbial activity of five faecal inocula from weaned pigs and one faecal inoculum from unweaned pigs in combination with 6 substrates. The substrates tested were negative control diet, corn, soybean meal, oligofructose (OF), ground chicory roots and a mixture (60% chicory pulp and 40% OF). The inocula used were derived from pigs fed either a corn-soy based diet without antibiotics (NCON), the NCON diet supplemented with oligofructose (OF), a mixture of chicory pulp (40%) and OF (60%) (MIX), ground chicory roots (CHR) or the NCON diet supplemented with antibiotics (PCON). The cumulative gas production measured fermentation kinetics and end products, such as total gas production, ammonia and volatile fatty acids, were also determined. Both the substrate and the inoculum significantly affected the fermentation characteristics. The cumulative gas production curve showed that different substrates caused more differences in traits of fermentation kinetics than the different inocula. Inocula of weaned pigs gave a significantly higher VFA production compared to the inoculum from unweaned animals, whilst the rate of fermentation and the total gas produced did not differ. OF showed the highest fermentation kinetics and the lowest NH3, pH and OM loss compared to other substrates. It was concluded that the microbial activity was significantly affected by substrate and inoculum. Inoculum from weaned pigs had more potential for microbial fermentation of the carbohydrate ingredients and oligofructose than that of unweaned pigs. A combination of high and low polymer inulin may be more beneficial to the gut ecosystem than using high- or low-polymer inulin alone. (Key Words : Oligofructose, Fermentation, Pig, Faeces, In vitro)

Global competition for high standard feed-food resources between man and livestock, such as industrial broilers, is a concerning problem. In addition, the low productivity of scavenger chickens in developing countries leaves much to be desired. Changing the ingredients, and therefore, the nutrient composition of feed intake by commercial fed as well as scavenger chickens seems like an obvious solution. In this study, the ability of four broiler chicken breeds to perform on a commercial versus a scavenger diet was tested. The four broiler breeds differed genetically in growth potential. A significant (P < 0.01) negative effect of the scavenger diet on the bodyweight of the fast growing breeds was found and this effect decreased with decreasing growth rate in the other breeds. These differences in bodyweight gain could not be explained by differences in nutrient digestibility but were caused by the lack of ability of the fast growing breeds to increase their feed intake sufficiently.

The intestinal microbiota is of great importance to our health and wellbeing. Modulation of the intestinal microbiota by exogenous and endogenous substrates can be expected to improve various physiological functions of our body, not just those in the intestine. Recently, new targets such as immune function and areas outside the colon have been considered to be influenced by the intestinal microbiota. Novel approaches might include the application of prebiotics in different combinations or the provision of nutrients to different bacterial groups and to different parts of the intestine.

The in vitro cumulative gas production technique can be used to assess microbial activity of a complex community, in relation to fermentation of a particular energy source. Therefore, in combination with an in vivo study to examine the effects of two different diets for weaning piglets, microbial activities of faeces were compared from animals on the two different diets. The two diets were: CHO diet [containing added fermentable carbohydrates, including sugarbeet pulp (SBP) and wheat starch (WST)], and control diet without any added fermentable carbohydrates. Neither diet contained antibiotics or extra added copper. Twenty-four piglets were selected from 12 litters (two per litter), weaned at 4 weeks of age (neither creep feeding nor any antibiotic treatment before and during the study), and introduced to one of the two diets. After 9 days on the diet, faecal samples were collected from selected animals, and tested for their activity in terms of gas production kinetics, and end-products such as volatile fatty acids, ammonia and dry matter disappearance of the two test substrates SBP and WST. The bacterial diversity was also analysed before and after in vitro fermentation using denaturing gradient gel electrophoresis analysis of amplified 16S rRNA genes. There were differences both in kinetics and end-products of the substrates. More interestingly, significant differences were detected between inocula, although mainly in terms of fermentation kinetics of the two substrates. With the CHO inoculum, SBP was fermented faster than with the control, while this effect was reversed for WST. Significantly higher diversity, as measured by DGGE fingerprint analysis, was detected in the microbial community enrichment on SBP as compared with WST at the end of fermentation. The difference between the kinetics of SBP compared with WST fermentation by faecal microbiota from the CHO diet fed piglets suggests better adaptation to SBP fermentation than to WST fermentation. The WST fermentation was more unexpected, given that a significant amount of starch is known to be fermentable by the small intestinal microbiota. It was concluded that the microbial community composition and activity in the GIT may be changed in response to diet, and that this change can be detected in vitro. Copyright © 2005 Society of Chemical Industry

Preventive approaches against cancer have not been fully developed and applied. For example, the incidence of some types of cancer, including colon cancer, is highly dependent upon lifestyle, and therefore, amenable to prevention. Among the lifestyle factors, diet strongly affects the incidence of colon cancer; however, there are no definitive dietary recommendations that protect against this malignancy. The association between diet-derived bioactives and development of colonic neoplasms will remain ill defined if we do not take into account: (1) the identity of the metabolites present in the colonic lumen; (2) their concentrations in the colon; and (3) the effect of the colonic contents on the function of individual bioactives. We review two approaches that address these questions: the use of fecal water and in vitro models of the human colon. Compared to treatment with individual diet-derived compounds, the exposure of colon cancer cells to samples from fecal water or human colon simulators mimics closer the in vitro conditions and allows for more reliable studies on the effects of diet on colon cancer development. The rationale and the advantages of these strategies are discussed from the perspective of a specific question on how to analyze the combined effect of two types of bioactives, butyrate and polyphenol metabolites, on colon cancer cells.