Dose-Dependent Effects of a Soluble Dietary Fibre (Pectin) on Food Intake,Adiposity, Gut Hypertrophy and Gut Satiety Hormone Secretion in Rats

Soluble fermentable dietary fibre elicits gut adaptations, increases satiety and potentially offers a natural sustainable means of body weight regulation. Here we aimed to quantify physiological responses to graded intakes of a specific dietary fibre (pectin) in an animal model. Four isocaloric semi-purified diets containing 0, 3.3%, 6.7% or 10% w/w apple pectin were offered ad libitum for 8 or 28 days to young adult male rats (n = 8/group). Measurements were made of voluntary food intake, body weight, initial and final body composition by magnetic resonance imaging, final gut regional weights and histology, and final plasma satiety hormone concentrations. In both 8- and 28-day cohorts, dietary pectin inclusion rate was negatively correlated with food intake, body weight gain and the change in body fat mass, with no effect on lean mass gain. In both cohorts, pectin had no effect on stomach weight but pectin inclusion rate was positively correlated with weights and lengths of small intestine and caecum, jejunum villus height and crypt depth, ileum crypt depth, and plasma total glucagon-like peptide-1 (GLP-1) and peptide tyrosine tyrosine (PYY) concentrations, and at 8 days was correlated with weight and length of colon and with caecal mucosal depth. Therefore, the gut’s morphological and endocrine adaptations were dose-dependent, occurred within 8 days and were largely sustained for 28 days during continued dietary intervention. Increasing amounts of the soluble fermentable fibre pectin in the diet proportionately decreased food intake, body weight gain and body fat content, associated with proportionately increased satiety hormones GLP-1 and PYY and intestinal hypertrophy, supporting a role for soluble dietary fibre-induced satiety in healthy body weight regulation.     

Development changes to gut microflora metabolism in mice.

Developmental changes in the activities of bacterial nitrate reductase, nitroreductase and beta-glucuronidase and their response to fermentable dietary fibre, were investigated in caecal contents from suckling mice (2-week-old) and in mice aged 4-24 weeks fed either a purified fibre-free diet or that diet supplemented with 5% (w/w) pectin. There was no apparent age-related trend common to the three enzymes studied. Nitrate reductase activity in the mice fed the fibre-free diet did not markedly alter with age. Pectin administration, however, was associated with a significant increase in nitrate reductase activity, particularly in 4-week-old mice. Nitroreductase activity exhibited an overall upward trend in mice from 2 to 12 weeks and thereafter decreased. Caecal beta-glucuronidase activity in mice increased sharply between 2 weeks and 4 weeks of age, thereafter not changing significantly until the 24th week. Pectin feeding had no consistent effect on activities either of nitroreductase or beta-glucuronidase. The changes in enzyme activities with age were not related to the concentration of bacteria in the caecum, which was highest in the 2-week-old mice. We conclude that the weaning is a period in which marked changes in caecal bacterial enzyme activities can occur.     

Isolation of rat intestinal crypt cells

A technique is presented which yields single cells and intact crypts in suspension from unfixed rat intestinal mucosal epithelium. Everted lengths of intestine were digested by 27 mM sodium citrate in phosphate-buffered saline (pH = 7.3) at 37 degrees C. Mucosal cells were dislodged by vibratory stress (hand vortexing) following incubation for prescribed intervals at 37 degrees C in 1.5 mM ethylenediamine tetraacetic acid (EDTA) and 0.5 mM dithiothreitol (dtt). Alkaline phosphatase determinations, phase microscopy, and in vivo and in vitro evaluations of tritiated thymidine ([3H]TdR) incorporation were performed on isolated intestinal cells. Data indicate that cells were sequentially derived from villus tip to crypt base as judged by cellular morphology, alkaline phosphatase activity/mg protein and radioactivity per microgram protein. Upon completion of the intestinal cell isolation assay, scanning electron microscopy of the remaining intestine revealed that approximately 95% of the crypt openings were vacant; the villi were totally denuded; the supporting structures, including the lamina propria, appeared intact. In vitro radiolabelling of intestinal cell fractions enriched with crypts revealed a linear incorporation of [3H]TdR from 0-60 min which was strongly influenced by the presence of foetal calf serum (FCS). Measurements of the compensatory response of the mucosa to resection of 70% of the small bowel indicated that the mucosal cell separation is capable of detecting alterations in crypt cell proliferation. Previously, such alterations were monitored by other methods utilizing microdissection procedures or stathmokinetic agents.     

Equine pre-caecal and total tract digestibility of individual carbohydrate fractions and their effect on caecal pH response

The working hypothesis was that a minor postprandial caecal pH decline would affect apparent total tract digestibility (ATTD) of the fibre fraction in horses and, hence, that soluble fibre would amplify fermentation and consequently increase ATTD of fibre. This study was a 4 × 4 Latin Square design with a sequence of 17 days adaptation to the ration followed by 8 sampling days. The feed rations consisted of only timothy hay (Group H), hay plus molassed sugar beet pulp combined with either whole oats (Group OB) or barley (Group BB) and hay plus loose chaff based concentrate (Group M). Four horses fitted with permanent caecal cannulas and collection harnesses were used. A pH electrode with logger was inserted through the cannula and caecal pH was recorded at 1 min intervals for 8 h. The mobile nylon bag technique was used to quantify pre-caecal loss (PCL) of individual feedstuffs. Fibre was analysed as dietary fibre (DF), non-starch polysaccharides, soluble non-cellulosic polysaccharides (S-NCP), insoluble non-cellulosic polysaccharide (I-NCP) and neutral detergent fibre. The ATTD of the S-NCP fraction was above 0.8, which was 60% higher than for the I-NCP fraction. The PCL of starch were 0.98 (oats) and 0.75 (barley). The BB diet lowered (p < 0.001) postprandial caecal pH more than the other diets and a significant correlation was found between the lowest pH and ATTD of I-NCP (r = ?0.66; p = 0.005). In conclusion, this study successfully measured the in vivo digestibility of individual fibre fractions and found that S-NCP was more digestible than the I-NCP, and that a single meal of unprocessed barley was sufficient to decrease caecal pH to such an extent that the fibre digestibility of the whole diet was negatively affected.     

Isolation of rat intestinal crypt cells

Abstract. A technique is presented which yields single cells and intact crypts in suspension from unfixed rat intestinal mucosal epithelium. Everted lengths of intestine were digested by 27 mM sodium citrate in phosphate-buffered saline (pH = 7.3) at 37°C. Mucosal cells were dislodged by vibratory stress (hand vortexing) following incubation for prescribed intervals at 37°C in 1.5 mM ethylenediamine tetraacetic acid (EDTA) and 0.5 mM dithiothreitol (dtt). Alkaline phosphatase determinations, phase microscopy, and in vivo and in vitro evaluations of tritiated thymidine ([³H]TdR) incorporation were performed on isolated intestinal cells. Data indicate that cells were sequentially derived from villus tip to crypt base as judged by cellular morphology, alkaline phosphatase activity/mg protein and radioactivity per μg protein. Upon completion of the intestinal cell isolation assay, scanning electron microscopy of the remaining intestine revealed that approximately 95% of the crypt openings were vacant; the villi were totally denuded; the supporting structures, including the lamina propria, appeared intact. In vitro radiolabelling of intestinal cell fractions enriched with crypts revealed a linear incorporation of [³H]TdR from 0–60 min which was strongly influenced by the presence of foetal calf serum (FCS). Measurements of the compensatory response of the mucosa to resection of 70% of the small bowel indicated that the mucosal cell separation is capable of detecting alterations in crypt cell proliferation. Previously, such alterations were monitored by other methods utilizing microdissection procedures or stathmokinetic agents.     

Gastrointestinal tract metabolism of young turkeys fed diets supplemented with pure nystose or a fructooligosaccharide mixture

In a four-week experiment on 60 7-day-old BUT-9 male turkeys the effects of dietary fructooligosaccharides (pure nystose and a fructooligosaccharide mixture) supplemented at 1 and 2%, were studied on ileal and caecal metabolism. The control carbohydrate was cellulose, added also at 1 or 2%. Each dietary treatment consists of 10 birds kept individually. The average degree of polymerisation of the nystose and oligofructose preparation amounted to 2.9 and 4.1, respectively. The addition of nystose significantly decreased the pH value and viscosity in the ileal contents compared with the cellulose treatment. On the other hand, the oligofructose preparation increased the activity of sucrase and lactase in the ileal mucosal by 30-60% and 33-47%, respectively. Both fructan preparations similarly acidified the caecal and colonic digesta (by 0.2-0.4 pH units) as well as diminished the activity of bacterial harmful beta-glucuronidase (by 24-40%), but only nystose caused an enlargement of the caeca and effectively reduced caecal ammonia concentration, especially at a higher dose. Oligofructose supplementation at 2% caused a 3.5-fold increase of bacterial activity of alph- and beta-galactosidase, while 2% nystose resulted in 1.7 and 3 times higher alpha- and beta-glucosidases activities, respectively. Compared to oligofructose, dietary nystose increased propionic and decreased butyric fermentation in caeca. Nystose and oligofructose preparations added at 2% reduced the triacylglycerol concentration in the serum in comparison to the addition of 2% cellulose by 46 and 25%, respectively. Beside the fact that dietary levels of supplementation were of great importance, the results indicated that even small difference in the length of carbohydrate chain may cause different physiological responses.     

Glucose-induced hyperaccumulation of cyclic AMP and defective glucose repression in yeast strains with reduced activity of cyclic AMP-dependent protein kinase.

Addition of glucose or related fermentable sugars to derepressed cells of the yeast Saccharomyces cerevisiae triggers a RAS-mediated cyclic AMP (cAMP) signal that induces a protein phosphorylation cascade. In yeast mutants (tpk1w1, tpk2w1, and tpk3w1) containing reduced activity of cAMP-dependent protein kinase, fermentable sugars, as opposed to nonfermentable carbon sources, induced a permanent hyperaccumulation of cAMP. This finding confirms previous conclusions that fermentable sugars are specific stimulators of cAMP synthesis in yeast cells. Despite the huge cAMP levels present in these mutants, deletion of the gene (BCY1) coding for the regulatory subunit of cAMP-dependent protein kinase severely reduced hyperaccumulation of cAMP. Glucose-induced hyperaccumulation of cAMP was also observed in exponential-phase glucose-grown cells of the tpklw1 and tpk2w1 strains but not the tpk3w1 strain even though addition of glucose to glucose-repressed wild-type cells did not induce a cAMP signal. Investigation of mitochondrial respiration by in vivo 31P nuclear magnetic resonance spectroscopy showed the tpk1w1 and tpk2w1 strains, to be defective in glucose repression. These results are consistent with the idea that the signal transmission pathway from glucose to adenyl cyclase contains a glucose-repressible protein. They also show that a certain level of cAMP-dependent protein phosphorylation is required for glucose repression. Investigation of the glucose-induced cAMP signal and glucose-induced activation of trehalase in derepressed cells of strains containing only one of the wild-type TPK genes indicates that the transient nature of the cAMP signal is due to feedback inhibition by cAMP-dependent protein kinase.     

Developmental changes to gut microflora metabolism in mice

W.E. BRENNAN-CRADDOCK, A.K. MALLETT, I.R. ROWLAND AND S. NEALE. 1992. Developmental changes in the activities of bacterial nitrate reductase, nitroreductase and β-glucuronidase and their response to fermentable dietary fibre, were investigated in caecal contents from suckling mice (2-week-old) and in mice aged 4–24 weeks fed either a purified fibre-free diet or that diet supplemented with 5% (w/w) pectin. There was no apparent age-related trend common to the three enzymes studied. Nitrate reductase activity in the mice fed the fibre-free diet did not markedly alter with age. Pectin administration, however, was associated with a significant increase in nitrate reductase activity, particularly in 4-week-old mice. Nitroreductase activity exhibited an overall upward trend in mice from 2 to 12 weeks and thereafter decreased. Caecal β-glucuronidase activity in mice increased sharply between 2 weeks and 4 weeks of age, thereafter not changing significantly until the 24th week. Pectin feeding had no consistent effect on activities either of nitroreductase or β-glucuronidase. The changes in enzyme activities with age were not related to the concentration of bacteria in the caecum, which was highest in the 2-week-old mice.
We conclude that the weaning is a period in which marked changes in caecal bacterial enzyme activities can occur.     

Levels of coenzyme A--glutathione mixed disulfide in Escherichia coli.

The pool of coenzyme A--glutathione mixed disulfide (CoASSG) rapidly increased 2.0 times in response to oxygen starvation and 1.5 times in response to glucose starvation but did not change following ammonia starvation. The increase in the CoASSG pool resulted from an increase in the CoASSG fraction of the CoA pool from 42 to 66--93%. Fluoride, cyanide, chloramphenicol, and rifampicin all caused similar increases. Aerobic growth on fermentable sugars resulted in CoASSG making up 40--55% of the CoA pool while growth on nonfermentable carbon sources or anaerobic fermentation resulted in CoASSG replacing acetyl CoA and free CoA to make up 85--95% of the CoA pool. The CoASSG:ATP ratio varied inversely with the growth rate in two groupings of carbon sources made up of either fermentable or nonfermentable molecules. Cultures grown aerobically on fermentable sugars exhibited a lower CoASSG:ATP ratio reflecting the lower proportion of CoASSG in the CoA pool.     

Effect of level of fibre on performance and digestion traits in growing rabbits

An experiment was conducted to determine the effects of level of fibre on production performance, gastrointestinal tract development, caecum fermentation and fibrolytic activity in weaner to 2-month-old New Zealand rabbits. One hundred weaner rabbits were allocated in individual cages for five treatments in which they were fed each diet with ADF at 130 (NDF, 274.1; ADL, 39.7; CP, 158.4; EE, 26.8; starch, 252.5 g/kg), 160 (NDF, 292.9; ADL, 41.9; CP, 160.3; EE, 25.6; starch, 249.4 g/kg), 190 (NDF, 312.4; ADL, 50.5; CP, 157.6; EE, 24.2; starch, 218.2 g/kg), 220 (NDF, 343.9; ADL, 51.2; CP, 158.5; EE, 22.2; starch, 217.6 g/kg), and 250 g/kg (NDF, 375.7; ADL, 66.0; CP, 156.9; EE, 22.4; starch, 220.8 g/kg) original matter basis, respectively. The results were: ADG and F/G deteriorated with increasing fibre (P<0.05); ADI increased with increasing dietary ADF (P=0.002). The stomach relative weight (g/kg LW), small intestine relative weight (g/kg LW), caecum weight, caecum content weight and caecum relative weight (g/kg LW) all increased with increasing ADF (P=0.003, 0.007, 0.002, 0.010 and <0.001, respectively). Quadratic effects of level of ADF on the villus height, villus height/crypt depth of the duodenum and the villus height/crypt depth of the jejunum were obtained; cubic effects of level of ADF on the crypt depth of the jejunum and the villus height/crypt depth of the ileum were obtained. Cubic effects of level of ADF on the pH, the butyric acid (% total VFA) and C2/C4 were obtained. The NH₃–N concentration dropped and the acetic acid (% total VFA) increased when the dietary ADF increased (P<0.001). The fibrolytic activity measured in the caecal contents increased when the dietary ADF increased (P<0.001). Providing a ration consisting mainly of clover meal, wheat straw and peanut hulls, the optimum dietary ADF content for weaner to 2-month-old growing meat rabbits was shown to be 190 g/kg feed.     

High-Fat Diet Reduces the Formation of Butyrate,but Increases Succinate,Inflammation, Liver Fat and Cholesterol in Rats,while Dietary Fibre Counteracts These Effects

Introduction

Obesity is linked to type 2 diabetes and risk factors associated to the metabolic syndrome. Consumption of dietary fibres has been shown to have positive metabolic health effects, such as by increasing satiety, lowering blood glucose and cholesterol levels. These effects may be associated with short-chain fatty acids (SCFAs), particularly propionic and butyric acids, formed by microbial degradation of dietary fibres in colon, and by their capacity to reduce low-grade inflammation.

Objective

To investigate whether dietary fibres, giving rise to different SCFAs, would affect metabolic risk markers in low-fat and high-fat diets using a model with conventional rats for 2, 4 and 6 weeks.

Material and Methods

Conventional rats were administered low-fat or high-fat diets, for 2, 4 or 6 weeks, supplemented with fermentable dietary fibres, giving rise to different SCFA patterns (pectin – acetic acid; guar gum – propionic acid; or a mixture – butyric acid). At the end of each experimental period, liver fat, cholesterol and triglycerides, serum and caecal SCFAs, plasma cholesterol, and inflammatory cytokines were analysed. The caecal microbiota was analysed after 6 weeks.

Results and Discussion

Fermentable dietary fibre decreased weight gain, liver fat, cholesterol and triglyceride content, and changed the formation of SCFAs. The high-fat diet primarily reduced formation of SCFAs but, after a longer experimental period, the formation of propionic and acetic acids recovered. The concentration of succinic acid in the rats increased in high-fat diets with time, indicating harmful effect of high-fat consumption. The dietary fibre partly counteracted these harmful effects and reduced inflammation. Furthermore, the number of Bacteroides was higher with guar gum, while noticeably that of Akkermansia was highest with the fibre-free diet.     

Modeling Cell Dynamics in Colon and Intestinal Crypts: The Significance of Central Stem Cells in Tumorigenesis

Colon and intestinal crypts have been widely chosen to study cell dynamics because of their fairly simple structures. In the colon and intestinal crypts, stem cells (SCs) are located at very bottom of the crypt, fully differentiated cells (FDs) are located in the top of the crypt, and transit-amplifying cells (TAs) are in the middle of the crypt between FDs and SCs. Recently, it has been discovered that there are two types of stem cells in the intestinal crypts: central stem cells (CeSCs) and border stem cells. To investigate dynamics of mutants in colon and intestinal crypts, we develop a four-compartmental stochastic model, which includes two SC compartments, and TAs and FDs compartments. We calculate the probability of the progeny of marked or mutant cells located at each of these compartments taking over the entire crypt or being washed out from the crypt. We found that the progeny of CeSCs will take over the entire crypt with a probability close to one. Interestingly, the progeny of advantageous mutant TAs and FDs will be washed out faster than disadvantageous mutants. Saliently, the model predicts that the time that the progeny of wild-type central stem cells will take over the mouse intestinal crypt is around 60 days, which is in perfect agreement with an experimental observation.     

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.     

Soluble Fermentable Dietary Fibre (Pectin) Decreases Caloric Intake,Adiposity and Lipidaemia in High-Fat Diet-Induced Obese Rats

Consumption of a high fat diet promotes obesity and poor metabolic health, both of which may be improved by decreasing caloric intake. Satiety-inducing ingredients such as dietary fibre may be beneficial and this study investigates in diet-induced obese (DIO) rats the effects of high or low fat diet with or without soluble fermentable fibre (pectin). In two independently replicated experiments, young adult male DIO rats that had been reared on high fat diet (HF; 45% energy from fat) were given HF, low fat diet (LF; 10% energy from fat), HF with 10% w/w pectin (HF+P), or LF with 10% w/w pectin (LF+P) ad libitum for 4 weeks (n = 8/group/experiment). Food intake, body weight, body composition (by magnetic resonance imaging), plasma hormones, and plasma and liver lipid concentrations were measured. Caloric intake and body weight gain were greatest in HF, lower in LF and HF+P, and lowest in the LF+P group. Body fat mass increased in HF, was maintained in LF, but decreased significantly in LF+P and HF+P groups. Final plasma leptin, insulin, total cholesterol and triglycerides were lower, and plasma satiety hormone PYY concentrations were higher, in LF+P and HF+P than in LF and HF groups, respectively. Total fat and triglyceride concentrations in liver were greatest in HF, lower in LF and HF+P, and lowest in the LF+P group. Therefore, the inclusion of soluble fibre in a high fat (or low fat) diet promoted increased satiety and decreased caloric intake, weight gain, adiposity, lipidaemia, leptinaemia and insulinaemia. These data support the potential of fermentable dietary fibre for weight loss and improving metabolic health in obesity.     

PROLIFERATION KINETICS IN EPITHELIUM OF GUINEA-PIG COLON III. DISTRIBUTION OF BLOOD CAPILLARIES ALONG THE CRYPT

The colonic blood vessels of the guinea-pig were infused with carmine-gelatine and the occurrence along the crypt of mucosal capillaries containing the dye was examined. The pattern of capillary distribution along the crypt is inhomogeneous but is similar for short, medium and long crypts. Capillaries occur least frequently at the bottoms of crypts, that is within the lowest 0.05% of the crypt length. The frequency rises significantly between 0.05 and 0.40 of the crypt length; this rise is accompanied by an increase in the number of DNA-synthesizing cells. A significant fall in capillary frequency along the crypt between 0.35 and 0.45 of crypt length coincides with the beginning of a decrease in the number of DNA-synthesizing cells. The highest frequency of capillaries occurs in the upper 0.20 of the crypt length and beneath the lining epithelium.     

Fiber-rich diets alter rat intestinal leukocytes metabolism

This study addressed the following question: What is the effect of fermentable and nonfermentable fiber-rich diets on intestinal immune cells' function and metabolism? For this purpose, weaning rats received, for 8 weeks, two types of fiber-enriched (30%) diets with different fermentable/nonfermentable fiber ratios, that is, oat bran (0.3) and wheat bran (0.14). The results of these two experimental groups were compared with those of the low-fiber control group having a 0.22 fermentable/nonfermentable fiber ratio. The total number and proportion of leukocytes in plasma, total number of cells in the lymphoid organs, lymphocyte proliferative activity and capacity of phagocytosis, hydrogen peroxide production, and adherence of macrophages were investigated. The activities of key enzymes of glycolysis and glutaminolysis, and of the Krebs cycle of lymphocytes from the mesenteric lymph nodes and macrophages from the intraperitoneal cavity were determined. The metabolic response of lymphocytes and macrophages from rats fed the three diets to Bacillus Calmette-Guérin-stimulus was also investigated. The number of lymphocytes in the mesenteric lymph nodes was lower in both fiber-rich diets than in the control but did not have any difference in the remaining lymphoid organs. Wheat bran caused a significant reduction in the phagocytosis capacity and adherence index of macrophages, whereas oat bran did not have a significant effect. The response of glucose and glutamine metabolism to Bacillus Calmette-Guérin-stimulus was not altered by the diets in lymphocytes, whereas in macrophages, the increase in glutaminase and hexokinase activities was abolished.     

Fermentable dietary fiber potentiates the localization of immune cells in the rat large intestinal crypts

Intestinal crypts are composed of a well-defined hierarchy of epithelial cells, and proliferating epithelial cells reside close to the bottom of the crypts-even in the large intestine. We investigated whether CD8(+)and CD4(+)intraepithelial lymphocytes (IELs) and CD161(+) natural killer (NK) cells localized in proliferating or differentiated epithelial region of cecum and colon. Both proliferating epithelial layer cells and the immune cells along the longitudinal crypt axis of the large intestine were measured histochemically. Dietary intervention revealed that the physiological localization of the immune cells in the longitudinal crypt axis depended on the immune cell type. CD8(+) IELs were preferentially located among differentiated epithelial cells. In contrast, CD161(+) NK cells were located adjacent to the epithelial cells at the bottom of crypt. Cecal crypts contained significantly larger numbers of CD8(+) IELs than did colonic crypts. However, there was only a minor population of CD4(+) IEL in the cecal and colonic epithelia. Some dietary fibers increased the densities of CD8(+) IELs and CD161(+) NK cells in the cecum, with the magnitude of response varying among the types of fiber. There was a significant relationship between SCFA and the localization of immune cells, especially CD8(+) IEL and CD161(+) NK cells, which are considered to be involved in the maintenance of epithelial homeostasis.     

Metabolic response to dietary fibre composition in horses

The hypothesis for this study was that a higher dietary proportion of soluble fibre would result in stable and constant plasma metabolite and regulatory hormone concentrations. The study was a 4×4 Latin Square design with a sequence of 17 days adaptation to the ration followed by 8 sampling days. The feed rations consisted of only timothy hay (H), hay plus molassed sugar beet pulp combined with either whole oats (OB) or barley (BB) and hay plus a loose chaff-based concentrate (M). Four horses were fitted with permanent caecal cannulas and liquid caecal content was withdrawn manually and blood was drawn from the jugular vein at 0, 3 and 9 h postprandial. The horses were exercised daily at medium level for about 1 h. Samples were analysed for short-chain fatty acids (SCFA) and metabolic traits. Caecal SCFA and propionic acid concentrations increased with increased dietary starch and soluble fibre. The diet highest in soluble fibre (M) resulted in the highest plasma glucose and insulin concentrations in the morning, which then remained stable and constant throughout the day. A strong interaction (P<0.01) between time and diet was measured for plasma urea, glucose, insulin and leptin. The greatest variations in plasma glycaemic and insulinaemic responses were associated with the cereal grain diets (OB and BB). There were indications of a negative energy balance, which was reflected in a significantly higher plasma β-hydroxybutyrate concentration and a numerically higher non-esterified fatty acid concentration. In conclusion, this study found that inclusion of soluble fibre resulted in increased total caecal SCFA and propionic acid concentrations. This consequently resulted in stable and constant plasma glycaemic and insulinaemic responses. Diets with a high content of soluble fibre provided enough energy for horses at medium work level.     

Biodiversity and fermentative activity of caecal microbial communities in wild and farm rabbits from Spain

In order to study the microbial caecal ecosystem of wild and domestic rabbits through the fermentation characteristics and concentration and diversity of bacterial and archaeal communities, caecal samples from sixteen wild rabbits (WR) were contrasted with two groups (n = 4) of farm rabbits receiving low (LSF) or high (HSF) soluble fibre diets from 28 (weaning) to 51 days of age. DNA was extracted for quantifying bacteria and Archaea by qPCR and for biodiversity analysis of microbial communities by DGGE. Samples from WR had lower caecal pH and ammonia and higher volatile fatty acids concentration than farm animals. Lower acetate and higher butyrate proportions were detected in WR. Bacterial and archaeal DGGE profiles were clearly different between wild and farm rabbits, and diet-affected population of farm rabbits. Similarity index of bacteria was lower than 0.40 among WR, and 0.52 among farm rabbits. In conclusion, caecal fermentation characteristics differ between wild and farm rabbits, which harbour clearly different bacterial and archaeal communities. In farm rabbits, diversity is influenced by the dietary level of soluble fibre.     

A METHOD FOR QUANTITATION OF PROLIFERATIVE INTESTINAL MUCOSAL CELLS ON A WEIGHT BASIS: SOME VALUES FOR C57BL/6

A method for determining the number of intestinal mucosal crypts, S cells, and total proliferative cells, on a weight basis has been presented. The number of crypts was obtained (following injection of tritiated thymidine) by dividing the disintegrations per minute (dpm) per mg intestine by the dpm per crypt. Multiplication of the number of crypts per mg by the number of labeled cells per crypt (determined radioautographically) resulted in the number of S cells per mg intestine. Division of the number of S cells per mg by the fraction of proliferative cells in S (obtained by cell cycle analysis) resulted in the number of proliferative cells per mg intestine. Values for duodenum, jejunum, and ileum of male C57BL/6 mice are given.