Postprandial triglyceride response in normolipidemic, hyperlipidemic and obese subjects - the influence of polydextrose, a non-digestible carbohydrate.

BACKGROUND: Three independent trials were conducted to evaluate postprandial triglyceride (TG) responses in subjects with different lipid metabolism. The effect of polydextrose (PDX), a soluble non-digestible carbohydrate, on postprandial response was also studied using practically relevant, high fat meal interventions. METHODS: A total of 19 normolipidemic (average BMI 24.1 kg/m(2)), 21 overweight/hyperlipidemic (average BMI 29.6 kg/m(2)) and 18 obese/non-diabetic subjects (average BMI 33.6 kg/m(2)) were included in the study. On two separate occasions all subjects ate two high-fat meals (4293 kJ, 36% from fat), one with PDX (either 12.5 g or 15 g) and one without PDX during placebo-controlled, double-blind, crossover and randomized trials. To obtain the triglyceride measurements venous blood samples were taken before the consumption of the test meal and five times afterwards, up to 6 h post-test meal. The triglyceride responses were modeled using a mixed-effects linear model. RESULTS: The key variables that explain the variation of the postprandial triglyceride response in the different subject groups were: baseline triglyceride concentration, time point, and PDX vs. placebo treatment (p < 0.05). The maximum postprandial TG concentration was more pronounced in hyperlipidemic group compared to normolipidemic (p < 0.001) or obese groups (p < 0.01). The modeled TG response analysis showed that irrespective of the study population PDX supplementation was one of the factors significantly reducing triglyceride response compared to the placebo treatment (p < 0.05). CONCLUSIONS: Subjects with elevated fasting triglyceride levels display exaggerated and prolonged postprandial triglyceride responses. PDX, a soluble non-digestible carbohydrate, may offer a dietary concept for reducing the postprandial triglyceride response after the consumption of a meal containing a high concentration of fat.

Postprandial effects of polydextrose on satiety hormone responses and subjective feelings of appetite in obese participants.

BACKGROUND: Dietary fibers are associated with enhanced satiety. However, the mechanism of different dietary fibers contributing to satiety-related gastrointestinal (GI) peptide release, especially in an obese population, is still poorly understood. Polydextrose (PDX), a water-soluble glucose polymer, has demonstrated its ability to reduce energy intake at a subsequent meal, but its mechanism of action requires further research. Also, there is limited evidence on its capacity to regulate subjective feelings of appetite. This study examines the effects of PDX on postprandial secretion of satiety-related GI peptides, short chain fatty acids (SCFAs), lactic acid, and subjective appetite ratings in obese participants. METHODS: 18 non-diabetic, obese participants (42.0 y, 33.6 kg/m2) consumed a high-fat meal (4293 kJ, 36% from fat) with or without PDX (15 g) in an acute, multicenter, randomized, double-blind, placebo-controlled and crossover trial. Postprandial plasma concentrations of satiety-related peptides, namely ghrelin, cholecystokinin (CCK), glucagon-like peptide 1 (GLP-1), and peptide YY (PYY), as well as SCFAs and lactic acid were assessed. GI peptide, SCFA and lactate concentrations were then modeled using a linear mixed-effects model.The subjective feelings of hunger, satisfaction, and desire to eat were evaluated using visual analogue scales (VAS), which were analyzed as incremental areas under the curve (iAUC) during the satiation and satiety periods. RESULTS: We found that PDX supplementation increased plasma GLP-1 levels more than the placebo treatment (P = 0.02). In the whole group, GLP-1 concentrations found in participants older than 40 years old were significantly lower (P = 0.01) as compared to those aged 40 years or less. There were no statistically significant differences in postprandial ghrelin, CCK, or PYY responses. The lactic acid concentrations were significantly (P = 0.01) decreased in the PDX group, while no significant changes in SCFAs were found. PDX reduced iAUC for hunger by 40% (P = 0.03) and marginally increased satisfaction by 22.5% (P = 0.08) during the post-meal satiety period. CONCLUSION: Polydextrose increased the postprandial secretion of the satiety hormone GLP-1 and reduced hunger after a high-fat meal. PDX also reduced the elevated postprandial lactic acid levels in plasma. Therefore, PDX may offer an additional means to regulate inter-meal satiety and improve postprandial metabolism in obese participants.

Treatment of bran containing bread by baking enzymes; effect on the growth of probiotic bacteria on soluble dietary fiber extract in vitro.

Different ways of treating bran by baking enzymes prior to dough making and the baking process were used to increase the amount of water-soluble dietary fiber (DF) in wheat bread with added bran. Soluble DF was extracted from the bread with water and separated from the digestible material with gastrointestinal tract enzymes and by solvent precipitation. The baking enzyme mixtures tested (xylanase and glucanase/cellulase, with and without lipase) increased the amounts of soluble arabinoxylan and protein resistant to digestion. The isolated fiber was used as a growth substrate for 11 probiotic and intestinal Bifidobacterium strains, for commensal strains of Bacteroides fragilis and Escherichia coli, and for potential intestinal pathogenic strains of E. coli O157:H7, Salmonella typhimurium, and Clostridium perfringens. Fermentation analyses indicated that the tested strains had varying capacity to grow in the presence of the extracted fiber. Of the tested probiotic strains B. longum species generally showed the highest ability to utilize the fiber extracts, although the potential pathogens tested also showed an ability to grow on these fiber extracts. In sum, the enzymes used to improve the baking process for high-fiber bread can also be used to produce in situ soluble fiber material, which in turn can exert prebiotic effects on certain potentially beneficial microbes.

Probiotic cheese containing Lactobacillus rhamnosus HN001 and Lactobacillus acidophilus NCFM® modifies subpopulations of fecal lactobacilli and Clostridium difficile in the elderly.

Aging is associated with alterations in the intestinal microbiota and with immunosenescence. Probiotics have the potential to modify a selected part of the intestinal microbiota as well as improve immune functions and may, therefore, be particularly beneficial to elderly consumers. In this randomized, controlled cross-over clinical trial, we assessed the effects of a probiotic cheese containing Lactobacillus rhamnosus HN001 and Lactobacillus acidophilus NCFM on the intestinal microbiota and fecal immune markers of 31 elderly volunteers and compared these effects with the administration of the same cheese without probiotics. The probiotic cheese was found to increase the number of L. rhamnosus and L. acidophilus NCFM in the feces, suggesting the survival of the strains during the gastrointestinal transit. Importantly, probiotic cheese administration was associated with a trend towards lower counts of Clostridium difficile in the elderly, as compared with the run-in period with the plain cheese. The effect was statistically significant in the subpopulation of the elderly who harbored C. difficile at the start of the study. The probiotic cheese was not found to significantly alter the levels of the major microbial groups, suggesting that the microbial changes conferred by the probiotic cheese were limited to specific bacterial groups. Despite that the administration of the probiotic cheese to the study population has earlier been shown to significantly improve the innate immunity of the elders, we did not observe measurable changes in the fecal immune IgA concentrations. No increase in fecal calprotectin and ß-defensin concentrations suggests that the probiotic treatment did not affect intestinal inflammatory markers. In conclusion, the administration of probiotic cheese containing L. rhamnosus HN001 and L. acidophilus NCFM, was associated with specific changes in the intestinal microbiota, mainly affecting specific subpopulations of intestinal lactobacilli and C. difficile, but did not have significant effects on the major microbial groups or the fecal immune markers.

Gut microbiota of healthy elderly NSAID users is selectively modified with the administration of Lactobacillus acidophilus NCFM and lactitol.

Ageing changes gut microbiota composition and alters immune system function. Probiotics, prebiotics and synbiotics may improve the health status of elderly individuals by modifying the intestinal environment and the microbiota composition, and by stimulating the immune system. In this work, we studied the effects of synbiotic supplementation on the gut microbiota of healthy elderly volunteers. Fifty-one elders were randomly assigned to consume either a synbiotic dietary supplement or a placebo in addition to their usual diet for a 2-week period. The synbiotic product consisted of the probiotic Lactobacillus acidophilus NCFM and the prebiotic lactitol and was ingested twice a day, with a total daily dose of 10 g lactitol and 2 × 10(10) cells of probiotic bacteria. Before, during and after the intervention period fecal quantities of six phylogenetic bacterial groups were determined using quantitative PCR, and relative changes in total microbiota composition were assessed by percent guanine-plus-cytosine profiling. The microbiota profiles showed certain relative changes within the microbial community, and indicated an increase of bifidobacteria levels during synbiotic supplementation. Quantification by PCR confirmed the in changes in the microbiota composition; for example increases in total levels of endogenous bifidobacteria and lactobacilli were recorded. Throughout the 6-week study period there was a decrease unrelated to intervention in the Blautia coccoides-Eubacterium rectale bacterial group levels and Clostridium cluster XIVab levels, but this decrease appeared to be halted during the synbiotic intervention. In conclusion, putatively beneficial changes in microbiota were observed in the elderly subjects supplemented with the synbiotic product.

Intestinal mucosal adherence and translocation of commensal bacteria at the early onset of type 2 diabetes: molecular mechanisms and probiotic treatment.

A fat-enriched diet modifies intestinal microbiota and initiates a low-grade inflammation, insulin resistance and type-2 diabetes. Here, we demonstrate that before the onset of diabetes, after only one week of a high-fat diet (HFD), live commensal intestinal bacteria are present in large numbers in the adipose tissue and the blood where they can induce inflammation. This translocation is prevented in mice lacking the microbial pattern recognition receptors Nod1 or CD14, but overtly increased in Myd88 knockout and ob/ob mouse. This 'metabolic bacteremia' is characterized by an increased co-localization with dendritic cells from the intestinal lamina propria and by an augmented intestinal mucosal adherence of non-pathogenic Escherichia coli. The bacterial translocation process from intestine towards tissue can be reversed by six weeks of treatment with the probiotic strain Bifidobacterium animalis subsp. lactis 420, which improves the animals' overall inflammatory and metabolic status. Altogether, these data demonstrate that the early onset of HFD-induced hyperglycemia is characterized by an increased bacterial translocation from intestine towards tissues, fuelling a continuous metabolic bacteremia, which could represent new therapeutic targets.

Simulated colon fiber metabolome regulates genes involved in cell cycle, apoptosis, and energy metabolism in human colon cancer cells.

High level of dietary fiber has been epidemiologically linked to protection against the risk for developing colon cancer. The mechanisms of this protection are not clear. Fermentation of dietary fiber in the colon results in production of for example butyrate that has drawn attention as a chemopreventive agent. Polydextrose, a soluble fiber that is only partially fermented in colon, was fermented in an in vitro colon simulator, in which the conditions mimic the human proximal, ascending, transverse, and distal colon in sequence. The subsequent fermentation metabolomes were applied on colon cancer cells, and the gene expression changes studied. Polydextrose fermentation down-regulated gene ontology classes linked with cell cycle, and affected number of metabolically active cells. Furthermore, up-regulated effects on classes linked with apoptosis, with increased caspase 2 and 3 activity, implicate that polydextrose fermentation plays a role in induction of apoptosis in colon cancer cells. The up-regulated genes involved also key regulators of lipid metabolism, such as PPARα and PGC-1α. These results offer hypotheses for the mechanisms of two health benefits linked with consumption of dietary fiber, reducing risk of development of colon cancer, and dyslipidemia.

The effect of age and non-steroidal anti-inflammatory drugs on human intestinal microbiota composition.

Ageing has been suggested to cause changes in the intestinal microbial community. In the present study, the microbiota of a previously well-defined group of elderly subjects aged between 70 and 85 years, both non-steroidal anti-inflammatory drugs (NSAID) users (n 9) and non-users (n 9), were further compared with young adults (n 14) with a mean age of 28 years, by two DNA-based techniques: percentage guanine+cytosine (%G+C) profiling and 16S rDNA sequencing. Remarkable changes in microbiota were described with both methods: compared with young adults a significant reduction in overall numbers of microbes in both elderly groups was measured. Moreover, the total number of microbes in elderly NSAID users was higher than in elderly without NSAID. In 16S rDNA sequencing, shifts in all major microbial phyla, such as lower numbers of Firmicutes and an increase in numbers of Bacteroidetes in the elderly were monitored. On the genus level an interesting link between reductions in the proportion of known butyrate producers belonging to Clostridium cluster XIVa, such as Roseburia and Ruminococcus, could be demonstrated in the elderly. Moreover, in the Actinobacteria group, lower numbers of Collinsella spp. were evident in the elderly subjects with NSAID compared both with young adults and the elderly without NSAID, suggesting that the use of NSAID along with age may also influence the composition of intestinal microbiota. Furthermore, relatively high numbers of Lactobacillus appeared only in the elderly subjects without NSAID. In general, the lowered numbers of microbial members in the major phyla, Firmicutes, together with changes in the epithelial layer functions can have a significant effect on the colon health of the elderly.

Human intestinal microbiota and healthy ageing.

Earlier studies have indicated a decrease in anaerobes and bifidobacteria and a concomitant increase in enterobacteria in the intestinal microbiota with ageing. However, new data obtained with molecular techniques suggests decreased stability and increased diversity of the gut microbiota with advancing age. Further, no simple marker change in microbiota composition can be identified. Except for the reduced immune function, ageing itself may have relatively little effect on overall gastrointestinal function. Concomitant changes in nutrition, increased incidence of disease and corresponding use of medication with advancing age modify the composition of the microbial community of the gastrointestinal tract. This mini-review will focus on the recent findings on the gut microbiota of the elderly and on the potential benefits of probiotics, prebiotics and synbiotics.

Effect of overweight on gastrointestinal microbiology and immunology: correlation with blood biomarkers.

A cross-sectional study was carried out in order to compare intestinal microbiological and immunological biomarkers with blood glucose and lipids, satiety-related hormones and inflammatory biomarkers characterising differences between obese and normal weight subjects. Faecal and blood samples were obtained from twenty obese subjects with an average BMI of 32.9 kg/m2 and twenty normal weight subjects with an average BMI of 23.3 kg/m2. Blood insulin, TAG and leptin were significantly elevated, whereas concentrations of HDL and ghrelin were significantly decreased in the obese subjects. Inflammatory status in the obese subjects was characterised by a trend for elevated blood C-reactive protein (CRP; P = 0.06) and IL-6 (P = 0.02). The faecal microbial composition differed between the groups; less sulphate-reducing bacteria (P = 0.05) and a trend for less Bacteroides (P = 0.07) were measured for overweight subjects. Furthermore, an inverse correlation was demonstrated between faecal Bacteroides levels and waist circumference (P = 0.05). The faecal microbial metabolites differed between the groups; increased concentrations of branched-chain fatty acids, phenolics, valeric acid, di- and hydroxy acids were described in the obese subjects. No differences between the measured intestinal inflammatory biomarkers were detected. However, systemic inflammation (CRP and IL-6) was correlated with the faecal concentrations of phenolics and lactic acid (P < 0.05 and 0.05, and P < 0.01 and 0.05, respectively). In summary, weight-related differences were observed both in the intestinal microbial composition and its activity. The role of intestinal signals, such as phenolics and lactic acid in the development of weight-related problems, needs to be studied further.

Specific probiotics alleviate allergic rhinitis during the birch pollen season.

AIM: To investigate whether birch pollen allergy symptoms are linked with gut microbiota changes and whether probiotics have an effect on these. METHODS: Forty seven children with confirmed birch pollen allergy were randomized to receive either a probiotic combination of Lactobacillus acidophilus (L. acidophilus) NCFM (ATCC 700396) and Bifidobacterium lactis (B. lactis) Bl-04 (ATCC SD5219) or placebo in a double-blind manner for 4 mo, starting prior to onset of the birch pollen season. Symptoms were recorded in a diary. Blood samples were taken for analysis of cytokines and eosinophils. Fecal samples were analysed for microbiota components, calprotectin and IgA. Nasal swabs were taken for analysis of eosinophils. RESULTS: The pollen season induced a reduction in Bifidobacterium, Clostridium and Bacteroides which could not be prevented by the probiotic intervention. During the intervention, significantly higher numbers of B. lactis 11.2 x 10(7) +/- 4.2 x 10(7) vs 0.1 x 10(7) +/- 0.1 x 10(7) bacteria/g feces (P < 0.0001) and L. acidophilus NCFM 3.5 x 10(6) +/- 1.3 x 10(6) vs 0.2 x 10(6) +/- 0.1 x 10(6) bacteria/g feces (P < 0.0001) were observed in the probiotic group compared to the placebo group. During May, there was a tendency for fewer subjects, (76.2% vs 95.2%, P = 0.078) to report runny nose, while during June, fewer subjects, 11.1% vs 33.3%, reported nasal blocking in the probiotics group (P = 0.101). Concomitantly, fewer subjects in the probiotic group had infiltration of eosinophils in the nasal mucosa compared to the placebo group, 57.1% vs 95% (P = 0.013). Eye symptoms tended to be slightly more frequent in the probiotic group, 12.5 d [interquartile range (IQR) 6-18] vs 7.5 d (IQR 0-11.5) (P = 0.066) during May. Fecal IgA was increased in the placebo group during the pollen season; this increase was prevented by the probiotics (P = 0.028). CONCLUSION: Birch pollen allergy was shown to be associated with changes in fecal microbiota composition. The specific combination of probiotics used was shown to prevent the pollen-induced infiltration of eosinophils into the nasal mucosa, and indicated a trend for reduced nasal symptoms.

Influence of a combination of Lactobacillus acidophilus NCFM and lactitol on healthy elderly: intestinal and immune parameters.

With increasing age, a number of physiological changes take place which are reflected in immune and bowel function. These changes may relate to the commonly assumed age-related changes in intestinal microbiota; most noticeably a reduction in bifidobacteria. The current study aimed at modifying the intestinal microbiota with a potential synbiotic on selected immune and microbiota markers. Healthy elderly subjects were randomised to consume during 2 weeks either a placebo (sucrose) or a combination of lactitol and Lactobacillus acidophilus NCFM twice daily in a double-blind parallel trial. After the intervention, stool frequency was higher in the synbiotic group than in the placebo group and a significant increase in faecal L. acidophilus NCFM levels was observed in the synbiotic group, after baseline correction. In contrast to the generally held opinion, the study subjects had faecal Bifidobacterium levels that were similar to those reported in healthy young adults. These levels were, nevertheless, significantly increased by the intervention. Levels of SCFA were not changed significantly. Of the measured immune markers, PGE2 levels were different between treatments and IgA levels changed over time. These changes were modest which may relate to the fact that the volunteers were healthy. Spermidine levels changed over time which may suggest an improved mucosal integrity and intestinal motility. The results suggest that consumption of lactitol combined with L. acidophilus NCFM twice daily may improve some markers of the intestinal microbiota composition and mucosal functions.

Effect of four probiotic strains and Escherichia coli O157:H7 on tight junction integrity and cyclo-oxygenase expression.

Controversy exists as to whether contact between a probiotic bacterial cell and an epithelial cell in the gut is needed to confer beneficial effects of probiotics, or whether metabolites from probiotics are sufficient to cause this effect. To address this question, Caco-2 cells were treated with cell-free supernatants of four probiotics, Bifidobacterium lactis 420, Bifidobacterium lactis HN019, Lactobacillus acidophilus NCFM, Lactobacillus salivarius Ls-33, and by a cell-free supernatant of a pathogenic bacteria, Escherichia coli O157:H7 (EHEC). Tight junction integrity as well as expression of cyclo-oxygenases, which are prostaglandin-producing enzymes, were measured. Probiotic-specific as well as EHEC-specific effects on tight junction integrity and cyclo-oxygenase expression were evident, indicating that live bacterial cells were not necessary for the manifestation of the effects. B. lactis 420 cell-free supernatant increased tight junction integrity, while EHEC cell-free supernatant induced damage on tight junctions. In general, EHEC and probiotics had opposite effects upon cyclo-oxygenase expression. Furthermore, B. lactis 420 cell-free supernatant protected the tight junctions from EHEC-induced damage when administered prior to the cell-free supernatant of EHEC. These results indicate that probiotics produce bioactive metabolites, suggesting that consumption of specific probiotic bacteria might be beneficial in protecting intestinal epithelial cells from the deleterious effects of pathogenic bacteria.

Lactobacillus acidophilus 74-2 and butyrate induce cyclooxygenase (COX)-1 expression in gastric cancer cells.

Cyclo-oxygenase (COX) profile predicts prognosis of gastric cancer; COX-2 positive tumors are more often aggressive, and COX-2 suppression is protective against gastric cancer. In contrast, COX-1 suppression is harmful to the intestinal mucosa. The COX-1, COX-2, and COX-1ir expression profiles were measured with real-time PCR in primary (AGS) and metastatic (NCI-N87) gastric adenocarcinoma cell lines treated with butyrate, hyperosmolar medium, and, in the case of NCI-N87, cell-free supernatants of probiotic bacteria Lactobacillus acidophilus 74-2 and Bifidobacterium lactis 420. The cell lines showed differences in the profile when treated with either hyperosmolar medium or butyrate. In NCI-N87 COX-2 expression was higher but only COX-1 expression was significantly upregulated by butyrate. Similarly to butyrate, the cell-free supernatant of L. acidophilus 74-2 upregulated COX-1, while COX-2 expression remained unchanged. COX-1ir, including COX-3, was upregulated by probiotics and osmotic stress. In conclusion, consumption of L. acidophilus 74-2 could be beneficial for the expression of cytoprotective COX-1.

The effect of ageing with and without non-steroidal anti-inflammatory drugs on gastrointestinal microbiology and immunology.

Elderly individuals are more susceptible to gastrointestinal problems such as constipation than young adults. Furthermore, the common use of non-steroidal anti-inflammatory drugs (NSAID) among the elderly is known to further increase such gastrointestinal ailments. To describe the specific changes in elderly, intestinal microbes, their metabolites and immune markers were measured from faecal samples obtained from fifty-five elderly individuals (aged 68-88 years), using either NSAID or not, and fourteen young adults (aged 21-39 years). The faecal DM content increased with age but was significantly lower among the elderly NSAID users. The microbial metabolism was especially influenced by NSAID use and/or ageing, although fewer changes were observed in the composition of the microbial community, whilst the level of aerobes was increased in the elderly and the level of Clostridium coccoides-Eubacterium rectale reduced in the elderly NSAID users as compared with young adults. An increase in the concentrations of some branched SCFA and l-lactate but a decrease in some major SCFA concentrations were observed. Evidently, the decreased defecation frequency in the elderly directed colonic fermentation toward an unfavourable microbial metabolism but this was partially offset by the use of NSAID. Irrespective of the use of NSAID, the elderly subjects had significantly lower concentrations of faecal PGE2 than the young adults, reflecting possibly a reduced immune response. According to the present study more attention should be paid to the development of dietary products that seek to enhance bowel function, saccharolytic fermentation and immune stimulation in the elderly population.

Effect of prebiotic supplementation on a probiotic bacteria mixture: comparison between a rat model and clinical trials.

The effects of a probiotic mixture (PRO), supplemented with either galacto-oligosaccharide (GOS) or polydextrose (PDX), on cell numbers of lactic acid bacteria (LAB) and bifidobacteria (BIF) were studied in conventional rats and healthy human subjects. In rats the baseline BIF cell numbers were below the detection limit and were increased by the 2-week GOSPRO intervention. In contrast baseline LAB numbers in rats were high and not affected by the treatments. The human study consisted of two independent but concurrent trials; both started with PRO followed by GOSPRO or PDXPRO periods. In the human subjects variation in numbers of BIF and LAB were high. The GOSPRO group exhibited high counts of faecal LAB and BIF at the start and showed little or no effects of the interventions. In contrast, the PDX group had low faecal LAB and BIF numbers at the start and clearly increased cell numbers of BIF after the PDXPRO period, and LAB after the PRO and PDXPRO period, compared with the run-in period. We propose here that responses to pro- and prebiotics are dependent on baseline numbers of LAB and/or BIF, and that the conventional rat model does not predict well the treatment responses in humans. The survival of PRO was presumably enhanced by the use of prebiotic supplementation and advocates the use of particular combinations of pro- and prebiotics.

The effect of cocoa and polydextrose on bacterial fermentation in gastrointestinal tract simulations.

Effects of cocoa mass and supplemented dietary fiber (polydextrose) on microbial fermentation were studied by combining digestion simulations of stomach and small intestine with multi-staged colon simulations. During the four phases of digestion, concentrations of available soluble proteins and reducing sugars reflected in vivo absorption of nutrients in small intestine. In colon simulation vessels, addition of polydextrose to digested cocoa mass significantly increased concentrations of total short-chain fatty acids and butyric acid, from 103 to 468 mM (P<0.01) and from 12 to 22 mM (P<0.01), respectively. Long-chain fatty acid concentrations (decreasing from 1,222 to 240 mM) were mainly affected by the presence of digested cocoa mass. Cocoa mass with or without polydextrose addition significantly decreased production of cadaverine (P<0.02) and branched-chain fatty acids compared to control during colon simulations. Results indicate beneficial effects on metabolism of colonic microbiota after digestion of cocoa mass, and even more so with polydextrose addition.

Effect of polydextrose on intestinal microbes and immune functions in pigs.

Dietary fibre has been proposed to decrease risk for colon cancer by altering the composition of intestinal microbes or their activity. In the present study, the changes in intestinal microbiota and its activity, and immunological characteristics, such as cyclo-oxygenase (COX)-2 gene expression in mucosa, in pigs fed with a high-energy-density diet, with and without supplementation of a soluble fibre (polydextrose; PDX) (30 g/d) were assessed in different intestinal compartments. PDX was gradually fermented throughout the intestine, and was still present in the distal colon. Irrespective of the diet throughout the intestine, of the four microbial groups determined by fluorescent in situ hybridisation, lactobacilli were found to be dominating, followed by clostridia and Bacteroides. Bifidobacteria represented a minority of the total intestinal microbiota. The numbers of bacteria increased approximately ten-fold from the distal small intestine to the distal colon. Concomitantly, also concentrations of SCFA and biogenic amines increased in the large intestine. In contrast, concentrations of luminal IgA decreased distally but the expression of mucosal COX-2 had a tendency to increase in the mucosa towards the distal colon. Addition of PDX to the diet significantly changed the fermentation endproducts, especially in the distal colon, whereas effects on bacterial composition were rather minor. There was a reduction in concentrations of SCFA and tryptamine, and an increase in concentrations of spermidine in the colon upon PDX supplementation. Furthermore, PDX tended to decrease the expression of mucosal COX-2, therefore possibly reducing the risk of developing colon cancer-promoting conditions in the distal intestine.

Effects of lactose on colon microbial community structure and function in a four-stage semi-continuous culture system.

A semi-continuous four-channel colon simulator was used to study the effects of lactose for the first time on the growth and fermentation dynamics of colonic microbiota. In six separate simulations, lactose supplementation increased the total SCFA concentration by 3-5 fold as compared with the baseline in the respective vessels. The total bacterial density was inversely correlated with lactic acid production (P = 0.003), while production of butyrate (P = 0.007) and propionate (P = 0.02) correlated with higher numbers of bacteria. A major shift in the microbial community structure in the lactose supplemented vessels was demonstrated by bacterial genomic %G+C-profiling of the total population, where lactose supplementation induced a clearly dominant peak in the bifidobacteria prominent area, %G+C 60-65. The transient shift to increased numbers of bifidobacteria (23-27%) of all bacteria in the first two vessels was also confirmed by the bifidobacteria-specific QPCR-method. In conclusion, lactose produced dramatic changes in microbiota composition and activity as compared with the baseline fermentation.

In vitro effects on polydextrose by colonic bacteria and caco-2 cell cyclooxygenase gene expression.

A 4-stage colon simulator and a cell culture-based human intestinal epithelial function model were combined to study the effects of a soluble fiber, polydextrose (PDX), on intestinal microbes and mucosal functions relevant to the risk of colon cancer. We observed sustained degradation of PDX throughout the different stages of the model. The fermentation was characterized by gradual degradation of PDX, production of short-chain fatty acids, and no increasing in putrefactive markers. We observed less marked effects in the microbial densities. When we applied colon fermentation metabolites obtained from the simulators with PDX to Caco-2 colon cancer cell line, a significant dose-dependent decreasing effect on cyclooxygenase-2 (COX-2) and an increasing effect on COX-3 expression levels were observed. PDX concentration appeared not to have effect on the expression levels of COX-1. Overexpression of COX-2 and decreased expression of COX-1 have been suggested to be characteristics of colon cancer. The exact physiological role of COX-3, an intron-retaining splice variant of COX-1, is not known, but it is suspected to play a role in transcriptional regulation of COX-1 and COX-2. In vitro modulation of COX expression by colon microbial fermentation products of polydextrose offers an interesting starting point for further studies on possible risk-decreasing effect of PDX on the development of colon cancer.