Search

Background: Intestinal mucins escape digestion and enter the large bowel where they are degraded by the microbiota. To what extent and how mucins impact large-bowel physiology remain unclear., Objective: This study examined the large-bowel fermentation characteristics of mucins and mucin-derived O-glycan sugars and whether they affect gut immunity., Methods: Mucin secretion from the terminal ileum was determined from feces of ileorectostomized male Wistar rats (age 6 wk) fed an AIN76-based control diet (CD) for 15 d (experiment 1). Normal male Wistar rats (age 6 wk; 4 wk for experiment 4) were fed CD ± porcine stomach mucin (PM) at 6 or 12 g/kg diet, equivalent to 1.5 and 3 times the daily mucin secretion, for 14 d (experiment 2); CD ± N-acetylglucosamine (GlcNAc), fucose, or N-acetylneuraminic acid at 10 g/kg diet for 14 d (experiment 3); or CD ± PM (15 g/kg diet) or GlcNAc (10 g/kg diet) for 29 d (experiment 4). SCFAs, microbial composition, and cecal O-glycan content were assessed. IgA+ plasma cells and regulatory T cells and inflammatory cytokine expression in the cecum were evaluated (experiment 4)., Results: Daily mucin secretion corresponded to 43.2 μmol of O-glycans. Cecal O-glycan contents were comparable between CD- and PM-fed rats. PM-fed rats harbored more mucin-degrading bacteria. Cecal concentrations of acetate (+37%) and n-butyrate (+73%) were higher in 12-g/kg PM diet-fed rats versus CD (P < 0.05). Among O-glycan sugars, only GlcNAc produced higher n-butyrate concentrations (+68%) versus CD (P < 0.05), with increased numbers of butyrate-producing bacteria. GlcNAc increased the abundance of IgA+ plasma cells (+29%) and regulatory T cells (+33%) versus CD, whereas PM increased IgA+ plasma cells (+25%) (all P < 0.05). GlcNAc and PM decreased expression of Tnfa (-30%, -40%) and Ifng (-30%, -70%) versus CD (all P < 0.05)., Conclusions: Mucin-derived O-glycans act as endogenous fiber and maintain mucosal immune homeostasis via large-bowel SCFA production in rats., (Copyright © The Author(s) 2020.)

Background: Chronic kidney disease (CKD) is a worldwide health problem. Although the pathogenesis of CKD is still unclear, recent studies suggest that systemic inflammation caused by a dysregulated microflora and an impaired intestinal barrier is involved in CKD development., Objective: We investigated the effects of the fermentable dietary fibers (DFs), unmodified guar gum (GG), and partially hydrolyzed GG (PHGG) (i.e., substances with distinct viscosity characteristics) on CKD development, with a particular focus on colonic tight junction (TJ) barriers in mice., Methods: Male 7-wk-old ICR mice were fed an AIN-93G diet that contained 0.25% adenine for 2 wk to induce CKD. Mice fed adenine were then divided into 3 groups and fed the unsupplemented diet (CKD) or a diet containing 10% PHGG (CKD+PHGG) or GG (CKD+GG) for 3 wk. Control (CON) mice were fed an AIN-93G diet without adenine throughout the 5-wk experiment. Plasma urea concentration; the colonic TJ proteins zonula occludens (ZO) 1, ZO2, occludin, junctional adhesion molecule A (JAMA), and claudin isoforms; renal inflammatory cytokines tumor necrosis factor α (Tnfa), interleukin (Il ) 1β (Il1b), and Il6; and cecal short-chain fatty acids (SCFAs) and microflora were analyzed., Results: Compared with the CON, CKD+PHGG, and CKD+GG groups, the CKD group had a 2.2- to 4.4-fold higher plasma urea concentration and greater expression of inflammatory cytokine genes in the kidney, including Tnfa (4.4- to 48-fold), Il1b (4.6- to 56-fold), and Il6 (8.8- to 115-fold). The CON, CKD+PHGG, and CKD+GG groups had greater expression of colonic TJ proteins including ZO1 (2.9- to 3.7-fold), ZO2 (3.4- to 4.3-fold), occludin (3.0- to 3.3-fold), JAMA (4.4- to 5.4-fold), and claudin 7 (2.1- to 2.6-fold) and higher cecal SCFA (1.8- to 3.5-fold) and Lactobacillus (2.7- to 4.0-fold) concentrations than the CKD group., Conclusion: Supplemental feeding with fermentable DFs, such as GG and PHGG, might be effective for the prevention or management of CKD by restoring colonic barrier integrity and microflora composition, as shown in mice.

Background: The mechanism underlying transient increases in immunoglobulin (Ig) A concentrations in the cecal contents of rats fed fructo-oligosaccharide (FOS) is unclear. Objective: This study was designed to test whether increased IgA concentrations represent one aspect of the inflammatory response to increased permeability induced by FOS in the cecum. Methods: Seven-week-old male Wistar rats were fed a fiber-free semipurified diet (FFP) with or without supplemental FOS (60 g/kg diet) for 9 or 58 d [experiment (expt.) 1], 7 d (expt. 2), or 7 or 56 d (expt. 3). In addition to measuring IgA concentrations in cecal content, we assessed gut permeability, inflammatory responses (expt. 1), the number of IgA plasma cells in the cecal lamina propria, polymeric Ig receptor (pIgR) expression in the cecal mucosa (expt. 2), and the condition of the cecal mucus layer (expt. 3). Results: The cecal IgA concentration in the FOS-fed rats was 15-fold higher than that of the rats fed FFP for 9 d ( P < 0.05). Gut permeability estimated by urinary chromium-EDTA excretion, bacterial translocation to mesenteric lymph nodes, myeloperoxidase activity, and expression of inflammatory cytokine genes in the cecal mucosa was greater in the FOS-fed rats than in the rats fed FFP for 9 d. These effects were not observed in the rats fed FOS for 58 d (expt. 1). Accompanying the higher cecal IgA concentration, pIgR protein and the number of IgA plasma cells in the cecal mucosa were higher in the FOS-fed rats than in the rats fed FFP for 7 d (expt. 2). Destruction of the mucus layer on the epithelial surface, as evidenced by Alcian blue staining in the cecal sections, was evident in the rats fed FOS for 7 d, but the mucus layer appeared normal in the rats fed FOS for 56 d (expt. 3). Conclusions: These findings suggest that transient increases in cecal IgA concentrations induced by FOS in rats are associated with mucosal inflammation in response to increased gut permeability; these are presumably evoked by disruption of the cecal mucus barrier. The observed responses could contribute to the maturation of the gut immune system., Competing Interests: Author disclosures: TG, YS, TK, SH, NN, TT, KS, and TM, no conflicts of interest., (© 2017 American Society for Nutrition.)

Background: Whole grain consumption reduces the risk of major chronic diseases. It is not clear how whole grains exert their beneficial effects., Objective: The aim was to compare the physiologic effects of whole grain oat (WGO) flour with low bran oat (LBO) flour., Methods: Two AIN-93G-based diets were formulated with either WGO or LBO flour. Five-week-old male C57BL/6J mice were fed LBO (n = 11) and WGO (n = 13) diets for 8 wk. Cecal microbiota was profiled by pyrosequencing of the 16S ribosomal RNA gene. Data are reported as means ± SEMs or antilogs of the mean (mean - SEM, mean + SEM)., Results: The weight gain was 14.6% less in the WGO group during week 7 (P = 0.04). WGO improved insulin sensitivity as reflected by significantly lower plasma insulin [1500 (1370, 1650) ng/L vs. 2340 (2090, 2620) ng/L; P = 0.006], C-peptide (3980 ± 548 ng/L vs. 7340 ± 1050 ng/L; P = 0.007), and homeostasis model assessment-estimated insulin resistance (21.4 ± 2.3 vs. 34.7 ± 4.9; P = 0.03). Plasma total cholesterol was 9.9% less and non-HDL cholesterol was 11% less in the WGO group. A comparison of relative abundance indicated Prevotellaceae, Lactobacillaceae, and Alcaligenaceae families were 175.5% (P = 0.03), 184.5% (P = 0.01), and 150.0% (P = 0.004), respectively, greater in the WGO group and Clostridiaceae and Lachnospiraceae families were 527% (P = 0.004) and 62.6% (P = 0.01), respectively, greater in the LBO group. Cecal microbiota composition predicts 63.9% variation in plasma insulin and 88.9% variation in plasma non-HDL cholesterol., Conclusions: In mice, WGOs improved insulin sensitivity and plasma cholesterol profile compared with LBOs, and the effects were associated with the changes in cecal microbiota composition. Increasing WGO consumption may help improve insulin sensitivity and dyslipidemia in chronic diseases., (© 2015 American Society for Nutrition.)

Resistant starch (RS) is highly fermentable by microbiota in the colon, resulting in the production of SCFAs. RS is thought to mediate a large proportion of its health benefits, including increased satiety, through the actions of SCFAs. The aim of this study was to investigate the effects of a diet high in RS on luminal microbiota composition, luminal SCFA concentrations, and the expression of host genes involved in SCFA uptake, SCFA signaling, and satiety regulation in mucosal tissue obtained from small intestine, cecum, and colon. Twenty adult female pigs were either assigned to a digestible starch (DS) diet or a diet high in RS (34%) for a period of 2 wk. After the intervention, luminal content and mucosal scrapings were obtained for detailed molecular analysis. RS was completely degraded in the cecum. In both the cecum and colon, differences in microbiota composition were observed between DS- and RS-fed pigs. In the colon these included the stimulation of the healthy gut-associated butyrate-producing Faecalibacterium prausnitzii, whereas potentially pathogenic members of the Gammaproteobacteria, including Escherichia coli and Pseudomonas spp., were reduced in relative abundance. Cecal and colonic SCFA concentrations were significantly greater in RS-fed pigs, and cecal gene expression of monocarboxylate transporter 1 (SLC16A1) and glucagon (GCG) was induced by RS. In conclusion, our data show that RS modulates microbiota composition, SCFA concentrations, and host gene expression in pig intestine. Combined, our data provide an enhanced understanding of the interaction between diet, microbiota, and host.

The gut microbiota plays an essential role in intestinal immunity. Prebiotics, including galacto-oligosaccharides (GOS), are fermentable fibers that beneficially affect the host by stimulating the growth of specific microbial populations. We investigated the effect of GOS on colitis development and on immune variables in Smad3-deficient mice treated with the pathogen Helicobacter hepaticus. Mice were supplemented daily with 5000 mg GOS/kg body weight 2 wk prior to infection and 4 wk postinfection, a time period during which colitis severity peaks in this model. Mice (n = 4-8/treatment at each time) were killed preinfection (0 d) and at 3, 7, and 28 d postinfection to evaluate immune variables in the spleen and in mesenteric lymph nodes (MsLN) by flow cytometry. Colon and cecum samples were collected for histopathologic analysis. Fecal pellets (n = 8-9/treatment) were collected prior to infection to measure relative changes in Bifidobacterium ssp. and Lactobacillum ssp. by real-time PCR. GOS significantly reduced colitis severity in response to H. hepaticus (P < 0.0001). This was associated with a significant increase in the percentage of NK cells in the spleen (P < 0.001) and in MsLN (P < 0.001) at 3 d postinfection and a 1.5-fold increase in fecal Bifidobacterium ssp. (P = 0.003). GOS stimulated NK expression of CCR9, a chemokine receptor involved in lymphocyte trafficking to the gut preinfection (0 d) in the blood (P = 0.02), spleen (P = 0.033), and MsLN (P = 0.017). In addition, GOS stimulated colonic IL-15 production 3 d postinfection (P < 0.001). These data suggest that GOS reduces colitis by modulating the function and trafficking of NK cells and may provide a novel therapeutic strategy for individuals with inflammatory bowel disease.

There is a need for better understanding of the roles of dietary fats and fibers in colon cancer risk. We examined the effect of different dietary fiber and fat sources on an azoxymethane (AOM)-induced colon cancer in rats. In a 2 x 3 factorial design, rats were fed a semipurified diet containing soy-derived fiber (Fibrim), alpha-cellulose (Solkafloc) or resistant starch (RS; Hi-maize) at 10 g dietary fiber/100 g diet, combined with fish oil (FO) or sunflower seed oil (SSO) at 10 g/100 g diet, and lard added to all diets at 10 g/100 g, to provide a total of 20 g mixed fat/100 g diet. Sprague-Dawley rats (28 d of age) consumed diets for 4 wk and then two doses of AOM (15 mg/kg body) were administered 1 wk apart by subcutaneous injection. Rats were killed after 13 wk of consuming experimental diets. Colons were fixed in formalin and aberrant crypt foci (ACF) were quantified after staining. ACF counts were higher (+66%, P < 0.01) in rats fed SSO and RS, than in those fed alpha-cellulose and FO. Rats fed FO had 19% fewer ACF than those fed SSO (P < 0.05). alpha-Cellulose was associated with the highest cecal butyrate concentration (P < 0.001), the highest beta-glucuronidase specific activity (P < 0.001) and the lowest cecal water cytotoxicity (P < 0.001) relative to soy fiber- and RS-fed rats. There were inverse correlations between the number of ACF and cecal butyrate concentration (r = -0.33, P < 0.05) and between cecal water cytotoxicity and beta-glucuronidase activity (r = -0.70, P < 0.001). The greatest protection was associated with alpha-cellulose as the fiber source and FO as the fat source as measured by colon ACF numbers in rats.

Pectin is a soluble indigestible polysaccharide that stimulates cecal polyamine formation in rats. Bacteroides and fusobacteria, two numerically dominant bacterial population groups in the large intestine, were found to synthesize in vitro high amounts of spermidine and putrescine. The purpose of this study was to elucidate the effect of pectin on the polyamine production by defined bacterial species in vivo. Germfree male Wistar rats (n = 18) were randomly assigned to one of three treatments: (i) monoassociation with Bacteroides thetaiotaomicron + fiber-free diet; (ii) diassociation with B. thetaiotaomicron + Fusobacterium varium + fiber-free diet or (iii) diassociation with B. thetaiotaomicron + F. varium + fiber-free diet + 10% pectin. The cecal contents of monoassociated rats fed fiber-free diet contained large amounts (1.51+/-0.21 micromol/dry total cecum content) of spermidine which was the major polyamine. The cecum of diassociated rats fed the fiber-free diet contained even higher concentrations of spermidine (2.53+/-0.21 micromol/dry total cecum content) and also putrescine, which was now the dominant polyamine (putrescine 0.32+/-0.28 vs. 3.01+/-0.28 micromol/dry total cecum content; monoassociation vs. diassociation). Pectin consumption by diassociated rats led to an additional increase in the cecal concentrations of all polyamines: putrescine, spermidine and spermine were 40, 37 and 100%, respectively, higher in the diassociated rats consuming the pectin diet than in those consuming the pectin-free diet. Since the microbial counts in the cecum did not differ in the diassociated treatment groups, the elevated concentrations of polyamines observed in the pectin group must have been due to stimulated bacterial polyamine synthesis. The decline of individual polyamines from cecum to feces detected at the end of the study in all treatment groups and the high microbial counts in the cecum and in feces suggest that bacterial polyamines are absorbed in cecum and colon. Pectin stimulates intestinal microbes to synthesize large amounts of polyamines which may be utilized by the host.

We examined the role of the large intestine in zinc absorption in rats in three separate experiments. In the first experiment, we examined apparent zinc absorption in rats fed diets containing graded levels of zinc carbonate (0.015-0.535 mmol Zn/kg diet) and evaluated zinc status on the basis of the zinc concentrations in serum and several tissues. The zinc absorption and the serum zinc concentration increased with the zinc content of the diet up to 0. 153 mmol Zn/kg diet. Femoral and pancreatic zinc levels increased linearly up to 0.229 mmol Zn/kg diet. In the second experiment, a zinc carbonate suspension was administered into the cecum via an implanted cannula or into the stomach via an orogastric tube, and the rats were fed diets with or without a highly fermentable fiber, guar gum hydrolysate (GGH, 50 g/kg diet), with coprophagy prevention. The amount of instilled zinc corresponded to the amount of zinc ingested as a component of the diet by the rats of a control group, 0.229 mmol Zn/kg diet. Apparent absorption of cecally instilled zinc was approximately half that observed when zinc was administered into the stomach in both diet groups. Serum and femur zinc concentrations in the cecum-administered groups were approximately 50 and 25% lower, respectively, than those in rats administered zinc into the stomach. The results demonstrate that, in vivo, the absorptive efficiency in the large intestine is not sufficient to satisfy the rat's zinc requirement and does not change when the luminal environment is substantially altered by feeding GGH. In Experiment 3, the effects of cecocolonectomy on zinc absorption were examined in rats with gastric acid suppression. In the cecocolonectomized groups, serum zinc concentration was lower as a result of treatment with a proton pump inhibitor, omeprazole, than in vehicle-treated rats, but not in sham-operated groups. These findings suggest that the cecum and colon contribute to zinc absorption when absorption in the small intestine is impaired.

We examined the role of resistant protein and peptides in promoting cecal butyrate production in rats fed rapidly fermentable carbohydrates. Rats were fed diets containing raw potato starch (RPS, 200 g/kg diet) or fructooligosaccharide (FOS, 60 g/kg diet) with casein, soy or rice protein (250 g/kg diet) for 13 d. In rats fed RPS with casein, the major cecal organic acid was acetate (441 micromol), but lactate and succinate were also found in considerable amounts (324 micromol). Succinate was the major cecal organic acid (235 micromol) in rats fed FOS with casein. When rice protein was fed with RPS, the contribution of lactate was significantly lower and that of propionate tended to be higher (P < 0.1) than in rats fed casein. In rats fed rice protein with FOS, cecal butyrate and acetate were greater and cecal succinate was lower than in rats fed casein with FOS (P < 0.05). Despite the similar amounts of undigested protein in rice and soy proteins, soy protein did not similarly affect cecal butyrate in rats fed FOS or RPS. In another experiment, rats were fed diets containing high amylose cornstarch (HAS, 200 g/kg diet) with casein, casein + oligo-L-methionine (OM, 3 g/kg diet), soy protein, soy protein + OM (3 g/kg diet) or rice protein (250 g/kg diet) for 10 d. OM (digestibility, 31%) was substituted for the same amount of casein. Rats fed rice protein had greater cecal butyrate than rats fed casein (P < 0.05). OM supplementation to casein or soy protein increased cecal butyrate compared with rats fed casein or soy protein alone (P < 0.05). These data support our hypothesis that resistant protein and peptides promote cecal butyrate production and suggest that the differing potency of rice and soy proteins in promoting cecal butyrate production might be explained in part by the different amino acid composition of resistant protein.

Young male pigs consumed a diet of fatty minced beef, safflower oil, skim milk powder, sucrose, cornstarch and wheat bran. Starch provided 50% of total daily energy either as low amylose cornstarch, high amylose (amylomaize) cornstarch or as a 50/50 mixture of corn and high amylose starch. Neither feed intake nor body weight gain as affected by dietary starch. Final plasma cholesterol concentrations were significantly higher than initial values in pigs fed the 50/50 mixture of corn and high amylose starch. Biliary concentrations of lithocholate and deoxycholate were lower in pigs fed high amylose starch. Large bowel length correlated positively with the dietary content of high amylose starch. Concentrations of butyrate in portal venous plasma were significantly lower in pigs fed high amylose starch than in those fed cornstarch. Neither large bowel digesta mass nor the concentrations of total or individual volatile fatty acids were affected by diet. However, the pool of propionate in the proximal colon and the concentration of propionate in feces were higher in pigs fed amylose starch. Concentrations of starch were uniformly low along the large bowel and were unaffected by starch type. In pigs with cecal cannula, digesta starch concentrations were higher with high amylose starch than with cornstarch. Electron micrographic examination of high amylose starch granules from these animals showed etching patterns similar to those of granules obtained from human ileostomy effluent. It appears that high amylose starch contributes to large bowel bacterial fermentation in the pig but that its utilization may be relatively rapid.

The question addressed was whether the viscosity per se of dietary non-starch polysaccharides influences macronutrient digestion in broiler chickens. Water-soluble carboxymethylcellulose preparations of low (LCMC) or high viscosity (HCMC) were fed to broiler chickens (n = 10/group) from 21 to 35 d of age. The HCMC preparations reduced weight gain and raised water intake compared with LCMC. After the HCMC diet was fed, viscosity of the supernatant of small intestinal contents was significantly raised. The HCMC preparations raised the group mean ATP concentration in the digesta of duodenum plus jejunum, indicating that bacterial activity was increased. Consumption of HCMC depressed apparent fecal digestibility of lipids and nitrogen and also apparent ileal digestibility of starch. The dietary HCMC tended (P = 0.077) to reduce plasma triglyceride concentrations. After HCMC consumption, the weights of the small intestine and colon, without or with contents, were elevated. The data indicate that high viscosity of digesta in broiler chickens is associated with a reduced macronutrient digestion and impaired growth performance. Because the carboxymethylcellulose preparations were nonfermentable by fresh feces, we suggest that HCMC reduces macronutrient digestion by raising the viscosity of small intestinal contents, which is associated with enhanced bacterial fermentation due to accumulation of undigested material.

Certain indigestible oligosaccharides may benefit gastrointestinal tract health via fermentation and proliferation of desirable bacterial species. The purpose of this study was to elucidate effects of selected oligosaccharides on cecal and fecal short-chain fatty acid (SCFA) concentration, pH, total large bowel wet weight and wall weight, and gut microbiota levels in rats. Fifty male Sprague-Dawley rats were randomly assigned to one of five treatments: 1) control diet; 2) control diet + 5% microcrystalline cellulose (5% CC); 3) control diet + 5% CC + 6% fructooligosaccharides; 4) control diet + 5% CC + 6% oligofructose; or 5) control diet + 5% CC + 6% xylooligosaccharides. The control diet consisted of (dry matter basis) 20% protein, 65% carbohydrate, 10.5% fat, vitamin and mineral mixes. The duration of the study was 14 d. The oligofructose- and fructooligosaccharide-containing diets resulted in higher cecal butyrate concentrations compared with the control, cellulose and xylooligosaccharide diets. Generally, total cecal SCFA pools were higher while pH was lower from ingesting oligosaccharide-containing diets compared with control or cellulose diets. Cecal total weight and wall weight were higher from oligosaccharide consumption, whereas colonic total wet weight was higher for rats consuming xylooligosaccharides compared with other treatments; colon wall weight was unaffected by treatments. Cecal bifidobacteria and total anaerobes were higher whereas total aerobes were lower in rats fed oligosaccharide diets compared with those fed the control diet. Cecal lactobacilli levels were unaffected by treatment. Dietary incorporation of fermentable, indigestible oligosaccharides, by providing SCFA, lowering pH, and increasing bifidobacteria, may be beneficial in improving gastrointestinal health.

Evidence supports a pathogenic role of arachidonic acid-derived inflammatory mediators within the gastrointestinal tract of patients with inflammatory bowel disease. The purpose of this study was to assess the effects of an ulcerative colitis nutritional formula (UCNF) containing oligosaccharides, fish oil, gum arabic and antioxidants on plasma and colonic phospholipid fatty acid and prostaglandin profiles in pigs. Twenty-four growing barrows in two replications were equally randomized among four killing times (d 0, 7, 14 and 21), and one of two diets, a control and the UCNF. Diets contained comparable levels of protein, fat, and nonstructural carbohydrate and met 100% of the energy requirements of the pig. Intake and body weight were recorded daily while blood, urine and tissue samples were collected at time of kill. Within 1 wk of ingestion of the UCNF, the composition of plasma phospholipid fatty acids showed an increase in 20:5(n-3) and 22:6(n-3) (P < 0.0001) and a decrease in 20:4(n-6) and 18:2(n-6) (P < 0.0001). Similar effects were observed for the phospholipids in the colonic and cecal mucosa. Plasma prostaglandin E was unaffected by treatment, whereas thromboxane B2 and 6-keto-prostaglandin F1 alpha levels were significantly decreased after 7 d of UCNF ingestion. Ingestion of the UCNF resulted in a suppression in the synthesis of proinflammatory prostaglandins by cecal and colonic mucosal cells. Levels of colonic and cecal prostaglandin E, 6-keto-prostaglandin F1 alpha and thromboxane B2 were significantly decreased after 7 d of UCNF ingestion. These changes may have been mediated by rapid increases of (n-3) fatty acids into cellular phospholipids. Dietary supplementation with the UCNF may prove beneficial for patients with ulcerative colitis by modulating colonic prostaglandin synthesis.

Oxidized ethyl linoleate (OEL) was prepared by aeration at low temperature. Peroxide value (POV, mEq/kg lipid) of OEL was 1400; the major oxidized compounds were 9-hydroperoxy-cis, trans- and 13-hydroperoxy-trans, cis-octadecadienoate. Rats fed fiber-free or sugar-beet fiber (SBF, 100g/kg diet) diets were divided into three groups for each diet, and administered OEL (high OEL group), OEL diluted with ethyl linoleate (low OEL group, POV 700) and nonoxidized ethyl linoleate (EL group) through gastric tubes each day at 1400-1600 h (2.5 g/kg body wt) for 16 d. The relative wet weight, and DNA and protein contents of the cecal mucosa were higher in the high OEL groups than in the low OEL and EL groups in rats fed the fiber-free diet and in rats fed the SBF diet except for mucosal protein content. Spermidine concentration in cecal mucosa of rats fed the fiber-free diet was greater in the high OEL group than in the EL group. These results suggest that metabolism related to mucosal proliferation of the cecum was affected by the high dose of OEL. The total short-chain fatty acid (SCFA) concentration in the cecal contents of SBF-fed rats was 100% higher than the concentration in rats fed the fiber-free diet in the EL group, but the administration of low dose and high dose OEL lowered the SCFA concentration in fiber-fed rats to that of rats fed the fiber-free diet. Butyric acid concentration was markedly lowered by ingestion of OEL in a dose-dependent manner in rats fed the SBF diet. In contrast, the isobutyric acid concentration was higher in the OEL-treated groups than in the EL groups. We conclude that a low dose of OEL depresses cecal fermentation of dietary fiber with changes in SCFA composition, and that a high dose of OEL induces mucosal hypertrophy in the cecum. These data show that dietary oxidized lipids affect cecal metabolism and may be associated with colon cancer.

Two experiments were conducted to determine if supplementing soluble fiber (fructooligosaccharide, xylooligosaccharide or gum arabic) to a semi-elemental diet would beneficially change cecal and colonic microbiota populations and enhance epithelial cell proliferation. Experiments 1 and 2 used identical dietary regimens; mice and rats were given free access to a powdered semi-elemental diet. Animals were assigned to one of the four following treatment groups: control, no supplemental dietary fiber, fructooligosaccharide, xylooligosaccharide and gum arabic. Dietary fiber was supplied via drinking water at 30 g/L. In Experiment 1 populations of Bifidobacteria and total anaerobic flora were enumerated from the contents of the cecum and colon of weanling mice. Consumption of fructooligosaccharide increased (P < 0.05) the concentrations of Bifidobacteria and the ratio of Bifidobacteria to total anaerobic flora. In Experiment 2 tissue from the cecum and distal colon of weanling rats was examined for morphological changes of the mucosa. Consumption of xylooligosaccharide increased (P < 0.05) cecal crypt depth and labeling index relative to the other three treatments. Consumption of gum arabic and the control diet increased (P < 0.01) cecal proliferation zone. Consumption of xylooligosaccharide and the control diet increased (P < 0.01) cecal cell density (number of cells in a vertical-half of the crypt). Distal colonic crypt depth was greatest (P < 0.05) in controls and rats fed fructooligosaccharide, intermediate in those fed gum arabic, and smallest in those fed xylooligosaccharide. These results suggest that fructooligosaccharide effectively stimulates growth of Bifidobacteria and xylooligosaccharide supports a modest enhancement of cecal epithelial cell proliferation.

Calculations of prececal protein digestibility based on the stable isotope 15N and the chemical label homoarginine were compared, using casein doubly-labeled with both markers. After food was withheld overnight 24 miniature pigs were given a meal containing 15 g/100 g casein, including 4 g/100 g doubly-labeled protein, and chromic oxide as an indigestible marker. The intestine of eight animals each was removed 3, 6 or 12 h later, divided into 3 sections of equal length, and chyme was collected. Kjeldahl-N, 15N and homoarginine were determined in diet and chyme. Digestibility of casein in the distal third of the small intestine was 93.5 +/- 0.5% and 97.6 +/- 0.3% (P < 0.05) according to 15N and homoarginine label, respectively. Potential causes for this systematic difference were assessed. The data suggest that incorporation of 15N into endogenous proteins and re-entry into the intestinal lumen via secreta and desquamations is the major cause for the 4.2 +/- 0.4% lower digestibility based on the 15N as compared with the homoarginine labeling technique. A preferential occurrence of homoarginine in more easily digestible sections of the protein, faster release during the digestive process and absorption of homoarginine, or incorporation of 15N into proteins of intestinal bacteria are less likely to cause this difference.

Previous research indicated that prunes in the diet of men with hypercholesterolemia lowered plasma and LDL cholesterol concentrations. To further study lipid metabolism in response to ingesting prunes, we conducted an animal study to test the hypotheses that fiber extracted from prunes, compared with purified cellulose, lowers plasma and liver cholesterol in rats with diet-induced hyperlipidemia and that the response is dose dependent. Rats were randomly assigned to one of five experimental diet groups. Four of the diets contained cholesterol and cholic acid to induce hyperlipidemia. The fiber source in the hyperlipidemic diets was 6% cellulose, 3% prune fiber, 6% prune fiber or 3% pectin. The fifth group, the nonhyperlipidemic control, was fed a diet containing 6% cellulose without cholesterol or cholic acid. Rats consumed one of the five diets ad libitum for 28 d, then were killed after 16 h without food. Plasma, LDL and liver cholesterol concentrations were higher in the hyperlipidemic control than the nonhyperlipidemic control and lower in the groups fed diets containing pectin or prune fiber than in the hyperlipidemic control group. No differences in plasma or liver cholesterol concentrations were detected between groups fed either dose level of prune fiber or between groups fed 6% prune fiber and pectin. These results indicate that fiber extracted from prunes lowers plasma and liver cholesterol in hyperlipidemic rats, although a dose-dependent response was not detected.

Influences of galactosylsucrose and xylosylfructoside on body mass gain, the digestive tract mass and concentrations of organic acids such as acetic, propionic, butyric, lactic and succinic acid in the cecum were compared among rats fed a cholesterol-enriched fiber-free diet [cholesterol 6 + cholic acid 1.5 (g/kg)] containing either galactosylsucrose, xylosylfructoside or sucrose (100 g/kg) or the above fiber-free diet without test sugar (control) for 21 d. Body mass gain was greater in rats fed sucrose, but not in rats fed galactosylsucrose or xylosylfructoside, than in control rats. The mass of the small intestine and colon plus rectum was larger in rats fed xylosylfructoside than in control rats. Cecal contents and cecal tissue mass were heavier, water content of cecal contents was higher, and pH and ammonia concentration of cecal contents were lower in rats fed diets containing xylosylfructoside than in control rats. Galactosylsucrose had similar effects, although not all differences were significant. The concentration of hydrogen ion in cecal contents positively correlated to total cecal concentration of measured organic acids and to amount of cecal contents. Total concentration of measured organic acids in cecal contents positively correlated to cecal tissue mass. The estimated contribution of galactosylsucrose, xylosylfructoside and sucrose for body mass deposition were 0.19, -0.29 and 0.51 (g body mass gained/g sugar), respectively.

The effects of corn oil, guar gum and cellulose on mucosal proliferation were investigated in rats. Animals were allocated to three groups and fed a fiber-free diet or diets containing 100 g/kg of cellulose or guar gum. Each group was subdivided to receive corn oil at 40 or 80 g/kg. The crypt cell production rate (CCPR) was determined after 28 d. Consumption of guar gum or corn oil led to greater CCPR in the ileum and cecum. In a second experiment, animals were allocated to two groups and fed diets containing either cellulose or guar gum (100 g/kg). Each group was again subdivided to receive either corn oil (80 g/kg) or minimal lipid (linolenic acid, 10 g/kg). The trophic effect of guar gum occurred even in the low lipid-fed group, indicating that guar gum exerts a positive effect on cell turnover independently of any interaction with luminal lipid. However, the highest CCPR occurred in animals fed guar gum and corn oil. Postprandial enteroglucagon and gastrin concentrations were highest in animals fed both guar gum and corn oil. Thus, corn oil and guar gum exert independent trophic effects on the intestinal mucosa. The combination of effects led to a three- to four-fold increase in colon mucosal CCPR.

The digestive and metabolic effects of inulin (from chicory) were studied in rats adapted to semipurified diets containing 0, 5, 10 or 20% inulin (wt/wt). Moderate levels of inulin (5-10%) did not significantly affect food intake or body weight gain. Dietary inulin resulted in considerably greater cecal fermentation and a significantly greater intraluminal concentration of propionate (peaking at 58.4 mmol/L). A lower concentration of acetate (42.6 mmol/L) was observed in rats fed 20% inulin. Lactic fermentations were observed in rats fed the 10 or 20% inulin diets. The cecal pool of volatile fatty acids tended to reach a plateau in rats fed diets containing more than 10% inulin (up to 600-700 mumol), but volatile fatty acid absorption was a slightly hyperbolic function of the dietary inulin level. Butyrate absorption was proportionally lower than that of propionate. Inulin-containing diets induced an enlargement of the cecal pool of calcium, phosphate and (to a lesser extent) magnesium. There was also an enhanced absorption of these divalent cations. The cecal pool of bile acids was greater in rats fed inulin, and this oligosaccharide displayed a slight hypocholesterolemic effect, even in rats fed the 5% inulin diet. However, plasma triglycerides were depressed only in rats fed the 20% inulin diet. In conclusion, inulin seems very effective in promoting propionic fermentation and in enhancing the calcium content of the large intestine. However, high levels of inulin (greater than 10%) may affect growth in rats and lead to acidic (pH 5.65) cecal fermentation.

Absorption of the short-chain fatty acids (SCFA), acetate, propionate and butyrate, was studied in vivo by measuring their disappearance from the washed cecum of anesthetized rats. The SCFA seemed to be absorbed predominantly by simple passive diffusion, because there was linear absorption over a wide concentration range, and no evidence of competition or enhancement of absorption when more than one SCFA was present in the lumen. Absorption of these three SCFA was independent of chain length. Their absorption was clearly limited by the lipid membrane, because permeability through the unstirred water layers was much greater than through the lipid membrane. Absorption was unaffected by altering the pH of the luminal infusate between 5.4 and 7.4. Based on our observation that, during absorption, there were increases in pH, total CO2 and HCO3- but no change in pCO2, we suggest that SCFA are absorbed primarily via diffusion involving anionic exchange with HCO3-.

Mature male Sprague-Dawley rats were assigned to a 3 x 3 factorial experiment in which they were fed AIN-76A diets supplying 8, 16 and 32% of energy as protein and 12, 24 and 48% of energy as fat. During the 5 mo of feeding, 10 in vivo measurements of intracolonic pH were recorded on each rat with a flexible electrode. The pH ranged from 7.8 to 8.0 near the anus and declined to 7.4 to 7.5 at 12 cm from the anus. The mean percentages of dry matter in the contents of the colon, divided into three approximately equal segments, were as follows: proximal colon, 35; middle colon, 45; distal colon, 58. Ammonia concentrations in luminal fluid rose significantly with higher protein intake in the cecum, proximal colon and distal colon. The concentrations in the distal colon ranged from 39 to 74 mmol/L, depending upon protein intake. Thymidine incorporation by distal colon mucosal cells was higher in rats fed 32% of energy as protein and 48% of energy as fat compared with rats fed 8% of energy as protein and 12% as fat. The evidence suggests that increased intestinal cell proliferation in rats fed the high protein, high fat diet was due to greater concentrations of ammonia in the large intestine resulting from the high protein intake and greater concentrations of non-ionized ammonia resulting from the higher pH associated with increased fat intake. The actual determinations and calculations of ionized to non-ionized ammonia concentrations were compatible with the assumption that the large intestinal cells absorbed more ammonia at higher fat intakes.

Various sugars are known to stimulate intestinal calcium absorption by a mechanism that is still poorly understood. One of those, the disaccharide 4-gal-actosyl-sorbitol (lactitol), is only metabolized in the large intestine, where it is converted into acidic residues. We investigated the effect of this compound on net intestinal absorption and body retention of calcium in rats. Because dietary calcium is in a poorly absorbable form when it reaches the large intestine, attempts were made to modify the absorption of calcium present in the large intestine contents by administering lactitol. Net intestinal absorption and body retention of calcium were significantly increased by 2.5 g lactitol/(kg b.w.d) given by a gavage over a 7-d period. This was associated with a lower pH and a better availability to absorptive systems of calcium present in the large intestine contents. The results indicate that in rats lactitol given chronically increases net intestinal absorption of calcium with a subsequently more positive calcium balance.

These studies were conducted to determine to what extent dietary fibers, or related compounds such as lactulose or amylomaize starch, alter the flux of blood urea to the cecum and cecal absorption of ammonia in the rat. Cecal weight and pH values were not different among rats fed diets containing 10% lactulose, pectin or guar gum, or 25% amylomaize starch. However, the cecal wall weight was markedly higher with lactulose feeding than with the other polysaccharides, whereas volatile fatty acid concentrations were lower with lactulose. The fiber diets depressed cecal ammonia, particularly in the case of the amylomaize starch diet, whereas the lactulose diet enhanced the concentration of ammonia. Owing to cecal enlargement and enhanced blood flow, the diets containing fermentable carbohydrates promoted a higher flux of urea to the cecum and also higher ammonia absorption in spite of low concentrations of ammonia in the cecum. Lactulose led to particularly high transfer of urea and absorption of ammonia. High blood urea in rats fed a 50% casein diet led to a very high flux of urea to the cecum and, hence, to high ammonia absorption. The presence of polysaccharides amplified the flux of urea and ammonia in the cecum. This study suggests that oligosaccharides such as lactulose, although very effective for the acidification of the contents of the large intestine, may enhance cecal ammonia and its absorption. Polysaccharides such as amylomaize starch might show greater efficiency for lowering ammonia concentrations in the large intestine.

The ability of dietary fiber to modify the effects of a bile acid on permeability of the cecum and colon was studied. A cecal catheter, which permits administration of test materials to conscious, unrestrained rats over a period of several weeks, was designed. Rats were fed fiber-free diet or diets containing 20% bran or lignin. Permeability of the lower intestine was assessed indirectly by infusing polyethylene glycol (PEG) and measuring excretion of PEG in urine. Under these conditions there was little effect of diet on permeability of PEG. However, when sodium deoxycholate was infused with the PEG, permeability was increased in rats fed fiber-free and lignin diets. In contrast, rats fed the bran diet showed no such response to the bile acid. The interaction among type of dietary fiber, presence of bile acid and intestinal permeability may have important implications for the etiology of intestinal disease.

The long-term effects of fermentable fibers on colonic luminal pH and the epithelial cell cycle were compared in 50 male Sprague-Dawley rats fed either a defined basal fiber-free diet or the basal diet supplemented with 10% pectin, cellulose or guar or with 20% oat bran. After 8 mo, in vivo pH measurements revealed that acidification of luminal contents occurred in the cecum and in mid and distal colons of rats fed fiber-supplemented diets when compared with the fiber-free controls (P less than 0.05). Pectin and guar produced the greatest acidification of luminal contents, the largest increase in cecal surface area and the highest percentage of colonic cells in S-phase, as measured by flow cytometry. In the proximal colon of the pectin group 9.2 +/- 0.5% of cells were in S-phase (6.3 +/- 0.8% with the fiber-free group) (P less than 0.05) and in the distal colon of the guar group 10.9 +/- 1.4% were in S-phase (7.1 +/- 0.5% with the fiber-free group) (P less than 0.05). Even though the most fermentable fibers produced the greatest mitogenic response, there was no site-specific correlation between pH and mucosal cell growth except in the cecum. This suggests that fibers may act as colon cell growth factors by some mechanism other than extracellular pH changes.