The effects of transgalactosylated oligosaccharides on gut flora metabolism in rats associated with a human faecal microflora

The effect on various caecal bacteria and their metabolic activities of feeding diet containing transgalactosylated oligosaccharides (TOS) with or without Bifidobacterium breve (administered in the drinking water) was investigated in rats colonized with a human faecal microflora. TOS (5% w/w in diet) or TOS plus B. breve, given for 4 weeks, induced increases in caecal concentration of total anaerobic bacteria, lactobacilli and bifidobacteria, and decreases in numbers of enterobacteria. Caecal pH was significantly reduced by feeding TOS, as were the activities of β-glucuronidase and nitrate reductase. In contrast, β-glucosidase activity was increased in TOS-fed rats.
Dietary TOS was also associated with decreased conversion, by caecal contents, of the dietary carcinogen 2-amino-3-methyl-3H-imidazo[4, 5- f ] quinoline (IQ) to its genotoxic 7-hydroxy derivative.     

Dietary carbohydrates affect caecal fermentation and modify nitrogen excretion patterns in rats II. Studies with diets differing in protein quality

In 2 two‐factorial experiments, each conducted on 80 growing male rats, the effects of substituting 10% raw potato starch (PS), pectins (PEC), or cellulose (CEL) for wheat starch (WS) and the addition of tannic acid to WS (WSTA) were studied using diets differing in protein quality. Casein unsupplemented or supplemented with DL‐methionine and gluten unsupplemented or supplemented with lysine, methionine and tryptophan were used as protein sources in Experiment 1 and 2, respectively. Parameters indicative of caecal fermentation intensity (pH, acetic, propionic and butyric acid contents, digesta and tissue weight) and of protein metabolism (urea blood concentration, faecal and urinary nitrogen excretion) were determined. Ten‐day balance experiments were preceded by a 10‐day adaption period to respective carbohydrates given in a diet containing balanced protein.

In both experiments the type of carbohydrates affected the caecal concentration of individual and total SCFA and other parameters of fermentation intensity. Pectins and potato starch were fermented more intensively than cellulose. Faecal N excretion was increased by all carbohydrates substituted for cereal starch, and by tannic acid. Urinary excretion was greater on CEL than on PEC and WSTA containing casein and on other diets containing gluten. In both experiments urinary N excretion was the lowest on PEC diets. Protein quality had the greatest effect on apparent biological value and net protein utilization but all indices of protein utilization were also affected by carbohydrates. It is concluded that not only the amount of N excreted in faeces but also in urine is affected by the type and fermentability of carbohydrates.     

Different effects of difructose anhydride III and inulin-type fructans on caecal microbiota in rats

The effects of different kinds of inulin-type fructans on caecal microbiota were evaluated in rats. Four groups of male Wistar rats were fed either a control diet, or diets containing 5% inulin, 5% fructooligosaccharides (FOS), or 5% difructose anhydride III (DFAIII) for two weeks. In the DFAIII group, caecal propionate, butyrate, counts of bifidobacteria, and total anaerobes were lower than in the inulin group, while caecal propionate, succinate, counts of bifidobacteria, and total anaerobes were lower than in the FOS group. Compared to controls, in the DFAIII group the counts of clostridia in caecum were increased by 3 log units. However, this change was statistically not significant. There were no differences between inulin and FOS groups for the pool of short chain fatty acids in caecum and bacterial counts. Results indicate that DFAIII has different effects on caecal microbiota compared to inulin and FOS and that these differences are most likely due to the alpha(3-->2) bonds in DFAIII.     

Different effects of difructose anhydride III and inulin-type fructans on caecal microbiota in rats

Abstract

The effects of different kinds of inulin-type fructans on caecal microbiota were evaluated in rats. Four groups of male Wistar rats were fed either a control diet, or diets containing 5% inulin, 5% fructooligosaccharides (FOS), or 5% difructose anhydride III (DFAIII) for two weeks. In the DFAIII group, caecal propionate, butyrate, counts of bifidobacteria, and total anaerobes were lower than in the inulin group, while caecal propionate, succinate, counts of bifidobacteria, and total anaerobes were lower than in the FOS group. Compared to controls, in the DFAIII group the counts of clostridia in caecum were increased by 3 log units. However, this change was statistically not significant. There were no differences between inulin and FOS groups for the pool of short chain fatty acids in caecum and bacterial counts. Results indicate that DFAIII has different effects on caecal microbiota compared to inulin and FOS and that these differences are most likely due to the α(3→2) bonds in DFAIII.     

Effects of cellulose, carboxymethylcellulose and inulin fed to rats as single supplements or in combinations on their caecal parameters

We compared the effect of diets containing different nondigestible carbohydrates: cellulose (C), inulin (IN) and carboxymethylcellulose (CMC) as single supplements or in dietary combination on caecal physiology of rats. Sixty male Wistar rats (Rattus norvegicus) were divided into five groups and for 4 weeks were fed a casein diet with the compared carbohydrates (4% of diet) or a combination of IN+C or IN+CMC (both 4+4%). Diet intake and FCR index remained unaffected by the treatments, whereas IN improved the body weight gain of rats compared to CMC. Compared to C group, all diets containing IN and CMC decreased the caecal pH as well as enlarged the caecum, thus increasing the weights of contents and tissue, especially upon CMC treatment. Rats given carboxymethylcellulose (CMC and IN+CMC groups) had watery caecal digesta, and some of them suffered from diarrhoea. In the case of CMC, the caecal enlargement was due to tissue hypertrophy and digesta accumulation mostly in response to an increased bulk of contents. Unlike C+IN, the dietary combination of CMC- and inulin-enhanced fermentation in the caecum of rats, however the proportion of acetate, propionate and butyrate was less beneficial. Compared to CMC, inulin gave a higher concentration of SCFA, especially of butyrate and propionate. The action of inulin in the caecum of rats could be pronounced by dietary treatment combined with CMC.     

The effect of dietary mannitol on the caecal microflora and short-chain fatty acids in rats

Wistar rats fed 5%-mannitol diet showed significant decreases of viridans streptococci and bifidobacteria, and a remarkable increase of butyric acid concentration and a decrease of acetic acid in the caecal contents compared with the sucrose and basal diet groups. Stocky fusiform bacteria were increased in concentration in the rats on mannitol compared with the other groups.     

Dietary fructooligosaccharides alter the cultivable faecal population of rats but do not stimulate the growth of intestinal bifidobacteria

The effect of fructans on the cultivable faecal community of Bio Breeding rats fed diets containing 5% (m/v) food-grade fructooligosaccharide (FOS) was investigated. Culturing of faecal material using chicory inulin as the sole carbohydrate source revealed the presence of a greater diversity of inulin-utilizing bacterial species in FOS-fed rats as compared with the control rats, although both contained species which effectively utilized inulin. The majority of cultivable inulin-utilizing species fell within the Clostridium coccoides group and Clostridium leptum subgroup, some of which were related to previously cultured butyrate-producing bacteria from the intestines of various animals. The impact of FOS on the growth of the indigenous bifidobacteria community and three inulin-utilizing isolates was assessed using real-time polymerase chain reaction. While dietary FOS was found to stimulate the growth of all three inulin-utilizing isolates, no growth stimulation of the indigenous bifidobacteria community occurred over the duration of the feeding trial.     

Physiological effects of lactulose and inulin in the caecum of rats

A model experiment was performed on rats to evaluate the effect of partial or total substitution of saccharose (S) and cellulose (C) by preparations of lactulose and inulin on the development and metabolism of the caecum. In the experimental diets given to rats for 4 weeks, the examined preparations were administered either with an equivalent amount of cellulose (each at 4% of the diet) or as sole source of dietary fibre at 8% of the diet. Compared to the saccharose group cellulose had no effect, and low doses of lactulose and inulin in the diet increased to a medium extent the weight of the caecum wall and caecal digesta. The addition of lactulose and inulin at 8% increased significantly the content of caecal digesta (4.62 and 4.11 g/100g BW, respectively) and the weight of the caecal wall (1.10 and 0.86 g/100g BW, respectively), compared to the groups with saccharose and cellulose (0.73, 0.90 and 0.24, 0.28 g/100g BW, respectively). Cellulose and cellulose partially-substituted with lactulose and inulin caused an increase in the dry matter content of caecal digesta (26.5-27.5%), compared to other groups (21.8-22.8%). The administration of lactulose and inulin preparations was accompanied by a significant drop in pH (5.47-5.81), compared to the groups with cellulose or saccharose (6.83-6.91), and a decrease in the ammonia concentration in the caecal digesta, compared to the cellulose control (0.27-0.40 and 0.62 mg/g, respectively). The group with 8% lactulose was characterized by the highest activities of microbiological alpha- and beta-galactosidase and beta-glucosidase in the caecal digesta. Cellulose and both preparations significantly decreased the activity of beta-glucuronidase, compared to the saccharose group (0.39-0.89 and 1.52 U/g, respectively). The highest concentration of VFA in the caecal digesta was observed in the saccharose group (89.2 micromol/g), and the lowest concentration in the group where cellulose was totally substituted by lactulose and inulin (55.1 and 57.5 micromol/g, respectively). The total production of VFA in the caecum was fourfold higher with 8 % lactulose and inulin (254.7 and 236.4 micromol/100g BW, respectively) than in both controls groups (65.1 and 67.8 micromol/100g BW, respectively). The high dose of inulin and lactulose increased the share of propionic acid in the VFA profile (C2:C3:C4) compared to both control groups. When 4% inulin was added to the diet a significant increase of butyrate concentration in the caecum was observed.     

Arabinoxylans and inulin differentially modulate the mucosal and luminal gut microbiota and mucin-degradation in humanized rats

The endogenous gut microbiota affects the host in many ways. Prebiotics should favour beneficial intestinal microbes and thus improve host health. In this study, we investigated how a novel class of potential prebiotic long-chain arabinoxylans (LC-AX) and the well-established prebiotic inulin (IN) modulate the gut microbiota of humanized rats. Six weeks after axenic rats were inoculated with a human faecal microbiota, their colonic microbiota was similar to this inoculum (～ 70%), whereas their caecal microbiota was enriched with Verrucomicrobia and Firmicutes concomitant with lower abundance of Bacteroidetes. Moreover, different Bifidobacterium species colonized the lumen (B. adolescentis) and mucus (B. longum and B. bifidum). Both LC-AX and IN increased SCFA levels and induced a shift from acetate towards health-promoting propionate and butyrate respectively. By applying a high-resolution phylogenetic micro-array (HITChip) at the site of fermentation (caecum), IN and LC-AX were shown to stimulate bacterial groups with known butyrate-producers (Roseburia intestinalis, Eubacterium rectale, Anaerostipes caccae) and bifidobacteria (B. longum) respectively. Prebiotic administration also resulted in lower caecal abundances of the mucin-degrading Akkermansia muciniphila and potentially more mucin production by the host. Both factors might explain the increased caecal mucin levels for LC-AX (threefold) and IN (sixfold). These mucins were degraded along the colon, resulting in high faecal abundances of Akkermansia muciniphila for LC-AX and especially IN-treated rats. Finally, the microbial changes caused an adaptation period for the host with less weight gain, after which the host fine-tuned the interaction with this altered microbiota. Our results demonstrate that next to IN, LC-AX are promising prebiotic compounds by stimulating production of health-promoting metabolites by specific microbes in the proximal regions. Further, prebiotic supplementation shifted mucin degradation to distal regions, where mucin-degraders may produce beneficial metabolites (e.g. propionate by Akkermansia muciniphila), so that prebiotics may potentially improve gut health along the entire length of the intestine.     

Physiological effects of lactulose and inulin in the caecum of rats

A model experiment was performed on rats to evaluate the effect of partial or total substitution of saccharose (S) and cellulose (C) by preparations of lactulose and inulin on the development and metabolism of the caecum. In the experimental diets given to rats for 4 weeks, the examined preparations were administered either with an equivalent amount of cellulose (each at 4% of the diet) or as sole source of dietary fibre at 8% of the diet. Compared to the saccharose group cellulose had no effect, and low doses of lactulose and inulin in the diet increased to a medium extent the weight of the caecum wall and caecal digesta. The addition of lactulose and inulin at 8% increased significantly the content of caecal digesta (4.62 and 4.11?g/100g BW, respectively) and the weight of the caecal wall (1.10 and 0.86?g/100g BW, respectively), compared to the groups with saccharose and cellulose (0.73, 0.90 and 0.24, 0.28?g/100g BW, respectively). Cellulose and cellulose partially-substituted with lactulose and inulin caused an increase in the dry matter content of caecal digesta (26.5–27.5%), compared to other groups (21.8–22.8%). The administration of lactulose and inulin preparations was accompanied by a significant drop in pH (5.47–5.81), compared to the groups with cellulose or saccharose (6.83–6.91), and a decrease in the ammonia concentration in the caecal digesta, compared to the cellulose control (0.27–0.40 and 0.62?mg/g, respectively). The group with 8% lactulose was characterized by the highest activities of microbiological α- and β-galactosidase and β-glucosidase in the caecal digesta. Cellulose and both preparations significantly decreased the activity of β-glucuronidase, compared to the saccharose group (0.39–0.89 and 1.52?U/g, respectively). The highest concentration of VFA in the caecal digesta was observed in the saccharose group (89.2?μmol/g), and the lowest concentration in the group where cellulose was totally substituted by lactulose and inulin (55.1 and 57.5?μmol/g, respectively). The total production of VFA in the caecum was fourfold higher with 8 % lactulose and inulin (254.7 and 236.4?μmol/100g BW, respectively) than in both controls groups (65.1 and 67.8?μmol/100g BW, respectively). The high dose of inulin and lactulose increased the share of propionic acid in the VFA profile (C₂:C₃:C₄) compared to both control groups. When 4% inulin was added to the diet a significant increase of butyrate concentration in the caecum was observed.     

VARIATIONS IN THE INTESTINAL FLORA OF RATS ON PURIFIED DIETS. I. THE EFFECT OF PYRIDOXINE DEFICIENCY ON THE LEVEL OF COLI-AEROGENES ORGANISMS

SUMMARY: Presumptive counts were made of several groups of intestinal organisms in caecal and rectal contents of rats fed on purified diets with and without adequate levels of pyridoxine. In the absence of pyridoxine the numbers of coli-aerogenes bacteria were higher in the caecal contents of rats fed on a purified diet containing glucose. This might not have been so if the diet had contained starch instead of sugar.     

甘露寡糖对断奶仔猪肠道主要菌群的影响

选择健康的 35日龄断奶仔猪 4 8头 ,随机分为两组 ,每组 3圈 ,每圈 8头。一组为含 0 .5 %甘露寡糠 +基础日粮 ,另一组为基础日粮。试验开始第 7天 ,每组各随机选 6头仔猪 ,麻醉后剖腹取肠道内容物以测定盲肠和结肠大肠杆菌、乳酸杆菌及双歧杆菌浓度和 p H。试验结果表明 ,与对照组相比 ,甘露寡糖可以显著降低盲肠、结肠大肠杆菌浓度 (P<0 .0 5 ) ;同时显著提高盲肠乳酸杆菌和双歧杆菌浓度 (P<0 .0 5 ) ,但对结肠乳酸杆菌和双歧杆菌数影响不显著 (P>0 .0 5 )     

Evidence for the absence of participation of the microbial flora in the hypocholesterolemic effect of guar gum in gnotobiotic rats

Germ-free (GF) and heteroxenic (Hx) rats were given a hypocholesterolemic diet (Hyper) with or without 5% guar gum (GG) for 4 weeks. The GF and Hx rats fed GG diets showed a lower hepatic and plasmatic cholesterol level when compared with Hyper groups. This reduction of cholesterolemia was due to a decrease in the chylomicron + very low density lipoprotein (VLDL) fraction. The caecal and portal concentrations of propionate were 30% higher in Hx rats fed the GG diet than in Hx rats fed the Hyper diet. These results exclude the participation of the intestinal microflora in the hypocholesterolemic effect of GG, and show that guar gum nullifies the effect of the hypocholesterolemic diet in the GF rats.     

Effect of concentrate feeding sequence on equine hindgut fermentation parameters

Feeding a diet with a small amount of hay and a high proportion of concentrate given in large meals entails a risk of colic to horses as this can impact the hindgut microbial ecosystem. To counteract this potential negative effect, one feeding strategy is to modify the distribution sequence of concentrate and hay. The purpose of the present study was to assess the impact of feeding the concentrate meal before or with the hay meal on fermentative parameters postprandial variations in caecum and right ventral colon contents and faeces. We focused on the comparison of pH value and volatile fatty acid (VFA) concentrations between the three compartments. Four horses fitted with two cannulas in the caecum and right ventral colon were fed a 100% hay diet (H) for a 3-week period, and then a 40% concentrate–60% hay diet for two consecutive periods of 3 weeks each. Concentrate and hay were offered simultaneously in two equal meals at 0800 and 1700 h (HC1) during one period, or separately in two equal meals at 0800 and 1600 h for the concentrate and at 1000 and 1730 h for hay (HC2) during the second one. During the 2^nd week of each diet period (H, HC1, HC2), caecal, right ventral colonic and faecal samples were collected before (0) and at 2, 4, 6 and 8 h after the morning meal. pH values and VFA concentrations were measured. Adding concentrate to the hay was associated with a decrease of pH value in the caecum, right ventral colon and faeces and an increase of faecal VFA concentrations (P<0.01). An impact of the concentrate feeding sequence was observed on caecal pH and faecal propionate concentration (P<0.001) but none on the other parameters investigated. In the caecum and right ventral colon, pH values decreased and VFA concentrations increased within the range of 4 to 8 h after the morning meal. Our data suggested that the variation of faecal pH could be an indicator of those of the caecal and right ventral colonic pH with a delay of 2 h. The present study showed that hay and commercial pelleted could be safely distributed simultaneously, thus making easier every day horse feeding.     

The influence of incubation pH on the activity of rat and human gut flora enzymes

The activities of three bacterial biotransformation enzymes (beta-glucuronidase, beta-glucosidase, nitrate reductase) were determined in suspensions of rat caecal contents or human faeces over the pH range 6-8. All three enzymes were influenced by pH, as exemplified by beta-glucosidase activity which diminished as pH increased. In other instances the rat and human flora showed distinct profiles, with nitrate reductase activity undetectable in human faeces below pH 6.6, whereas the rat caecal flora displayed optimal reduction of nitrate around neutrality. The most pronounced host-species difference was found with beta-glucuronidase, which showed maximal activity at pH 6.0 in human faecal bacteria, while the rat caecal flora expressed greatest activity at pH 8.0. All three enzyme activities were associated with that fraction of rat caecal or human faecal material sedimented by centrifugation at 5000 g for 15 min, with little or no metabolism occurring in the 11,000 g supernatant fluid. The results demonstrate that pH has a pronounced effect on the enzymic activity of bacterial preparations from rat and human sources.     

Effect of inulin supplementation on selected gastric, duodenal, and caecal microbiota and short chain fatty acid pattern in growing piglets

We explored whether bifidobacteria and lactobacilli numbers and other selected bacteria in the upper intestine and the caecum of growing pigs were affected by diet and intake of inulin. Starting at two weeks after weaning (28 d) 72 pigs were fed two types of diets (wheat/barley (WB) or maize/gluten (MG)), without or with 3% inulin (WB + I, MG + I) for three and six weeks. Intestinal bacteria were quantified by fluorescence-in-situ-hybridization (n = 8/group). Duration of feeding had no effect on the variables tested, so data for both periods were pooled. Gastric total bacteria amounted to log(10) 7.4/g digesta. Bifidobacteria were detected in stomach and duodenum two weeks after weaning and disappeared thereafter. In jejunum and caecum bifidobacteria were present at a level of log(10) 7.0/g digesta. Inulin did not alter numbers of lactobacilli, bifidobacteria, enterococci, enterobacteria and bacteria of the Clostridium coccoides/Eubacterium rectale-group. Inulin disappearance in stomach plus jejunum was higher with the MG diet (73.7 vs. 60.7%, p = 0.013). Caecal acetate was lower in inulin-supplemented diets (p < 0.05) whereas propionate and butyrate were higher in pigs fed the WB diets (p < 0.05). With the WB diet total caecal short chain fatty acids concentration was higher which resulted in a lower pH value (p < 0.05).     

Effects of insoluble dietary fibre differing in lignin on performance, gut microbiology, and digestibility in weanling piglets

The study aimed to investigate the effects of insoluble dietary fibre differing in lignin content on performance and parameters of gut microbiology in 48 weanling piglets (8.5 kg mean body weight) fed common diets ad libitum. Fibre sources tested were wheat bran (low lignin) and pollen from Chinese Masson pine (Pinus massoniana) as model of fibre rich in lignin, added to diets as follows: no addition (control), 3.0% wheat bran, 1.27% pine pollen and 2.55% pine pollen. In colonic chyme, bacterial colony counts remained unaffected by treatment, but ammonia contents were reduced by fibre additions (up to -38%). The effects on ammonia were best explained by added cellulose and lignin. Fibre additions reduced apparent (faecal) digestibility of dry matter (DM) and crude protein (CP) up to -3.5 and -4.3% units with cellulose and lignin being the major determinants for changes in CP digestibility.     

The influence of incubation pH on the activity of rat and human gut flora enzymes

The activities of three bacterial biotransformation enzymes (β-glucuronidase, β-glucosidase, nitrate reductase) were determined in suspensions of rat caecal contents or human faeces over the pH range 6–8. All three enzymes were influenced by pH, as exemplified by β-glucosidase activity which diminished as pH increased. In other instances the rat and human flora showed distinct profiles, with nitrate reductase activity undetectable in human faeces below pH 6–6, whereas the rat caecal flora displayed optimal reduction of nitrate around neutrality. The most pronounced host-species difference was found with β-glucuronidase, which showed maximal activity at pH 6–0 in human faecal bacteria, while the rat caecal flora expressed greatest activity at pH 8–0. All three enzyme activities were associated with that fraction of rat caecal or human faecal material sedimented by centrifugation at 5000 g for 15 min, with little or no metabolism occurring in the 11000 g supernatant fluid. The results demonstrate that pH has a pronounced effect on the enzymic activity of bacterial preparations from rat and human sources.     

Elimination of murine viral pathogens from the caecal contents of mice by anaerobic preparation

The aim of this study was to investigate methods to eliminate pathogenic viral agents while preserving the 'normalizing' properties of the gut microflora of mice. Mouse hepatitis virus A59 (MHV), Reo 3 virus and Theiler GD VII virus were added to the caecal contents of 'normal' mice and following dilution, with or without subsequent culturing, given to germ-free mice. Four weeks later antibody titres against these and other viruses were determined. MHV and Theiler CD VII virus survived dilution but were eliminated during culturing. Reo-virus survived the 10(-1) dilution-culture step. All dilutions and dilution-cultures of caecal contents resulted in 'normalization' in germ-free recipients of the relative caecal weight, percentage faecal fusiform-shaped bacteria, faecal bile acids and colonization of small intestine by segmented filamentous bacteria.     

Effect of dietary chitosans with different viscosity on plasma lipids and lipid peroxidation in rats fed on a diet enriched with cholesterol

To investigate the effect of dietary chitosan on lipid metabolism, male SD (Sprague-Dawley) rats were fed a cholesterol-enriched diet containing 5% cellulose (CE), 5% chitosan (CCS; high viscosity), or 5% chitosan (FCS; low viscosity) for 4 weeks. The two types of chitosan with a comparable degree of deacetylation had a different molecular weight and intrinsic viscosity. Significantly (p < 0.05) lower plasma total cholesterol, LDL-cholesterol and VLDL-cholesterol concentrations were observed in the rats fed on the chitosan diets. In addition, chitosan significantly increased the fecal cholesterol and triglyceride contents. Although no significant difference in body weight was found among the dietary groups, the rats fed on the chitosan diets had lower relative liver weight when compared with those fed on the cellulose diet. Both of the chitosan groups had significantly lower liver total lipid and total cholesterol contents compared to the cellulose group, although the FCS group was less effective. The plasma and liver thiobarbituric acid reactive substances (TBAR) values were similar in the CE and FCS groups, while the CCS group had increased liver TBAR values. Although a significant increase in liver glucose-6-phosphate dehydrogenase activity was observed in the CCS group, no significant change was found in the FCS group. The observed influence of chitosans with different viscosity on the plasma lipid level, liver lipids and lipid peroxidation suggests that, while the hypocholesterolemic action of chitosans with different viscosity was similar, changes in the liver lipids and liver peroxidation status depended on their molecular weight when the deacetylation degree was comparable.