Improved Tol2-mediated enhancer trap identifies weakly expressed genes during liver and ß cell development and regeneration in zebrafish.

The liver and pancreas are two major digestive organs, and among the different cell types in them, hepatocytes and the insulin-producing ß cells have roles in both health and diseases. Accordingly, clinicians and researchers are very interested in the mechanisms underlying the development and regeneration of liver and pancreatic ß cells. Gene and enhancer traps such as the Tol2 transposon-based system are useful for identifying genes potentially involved in developmental processes in the zebrafish model. Here, we developed a strategy that combines a Tol2-mediated enhancer trap and the Cre/loxP system by using loxP-flanked reporters driven by ß cell- or hepatocyte-specific promoters and the upstream activating sequence (UAS)-driving Cre. Two double-transgenic reporter lines, Tg(ins:loxP-CFPNTR-loxP-DsRed; 10×UAS:Cre, cryaa:Venus) and Tg(fabp10:loxP-CFPNTR-loxP-DsRed; 10×UAS:Cre, cryaa:Venus), were established to label pancreatic ß cells and hepatocytes, respectively. These two double-transgenic lines were each crossed with the Tol2-enhancer trap founder lines to screen for and identify genes expressed in the ß cell and hepatocytes during development. This trap system coupled with application of nitroreductase (NTR)/metronidazole (Mtz)-mediated cell ablation could identify genes expressed during regeneration. Of note, pilot enhancer traps captured transiently and weakly expressed genes such as rab3da and ensab with higher efficiencies than traditional enhancer trap systems. In conclusion, through permanent genetic labeling by Cre/loxP, this improved Tol2-mediated enhancer trap system provides a promising method to identify transiently or weakly expressed, but potentially important, genes during development and regeneration.

Effects of two whole-grain barley varieties on caecal SCFA, gut microbiota and plasma inflammatory markers in rats consuming low- and high-fat diets.

Mixed-linkage ß-glucans are fermented by the colon microbiota that give rise to SCFA. Propionic and butyric acids have been found to play an important role in colonic health, as well as they may have extraintestinal metabolic effects. The aim of the present study was to investigate how two whole-grain barley varieties differing in dietary fibre and ß-glucan content affected caecal SCFA, gut microbiota and some plasma inflammatory markers in rats consuming low-fat (LF) or high-fat (HF) diets. Barley increased the caecal pool of SCFA in rats fed the LF and HF diets compared with those fed the control diet, and the effect was generally dependent on fibre content, an exception was butyric acid in the LF setting. Furthermore, whole-grain barley reduced plasma lipopolysaccharide-binding protein and monocyte chemoattractant protein-1, increased the caecal abundance of Lactobacillus and decreased the Bacteroides fragilis group, but increased the number of Bifidobacterium only when dietary fat was consumed at a low level. Fat content influenced the effects of barley: rats fed the HF diets had a higher caecal pool of acetic and propionic acids, higher concentrations of amino acids and higher amounts of lipids in the portal plasma and liver than rats fed the LF diets; however, less amounts of butyric acid were generally formed. Interestingly, there was an increase in the caecal abundance of Akkermansia and the caecal pool of succinic acid, and a decrease in the proportion of Bifidobacterium and the Clostridium leptum group. In summary, whole-grain barley decreased HF diet-induced inflammation, which was possibly related to the formation of SCFA and changes in microbiota composition. High ß-glucan content in the diet was associated with reduced plasma cholesterol levels.

Lipid metabolism regulation by dietary polysaccharides with different structural properties.

Lipid metabolism plays an important role in energy homeostasis maintenance in response to stress. Nowadays, hyperlipidemia-related chronic diseases such as obesity, diabetes, atherosclerosis, and fatty liver pose significant health challenges. Dietary polysaccharides (DPs) have gained attention for their effective lipid-lowering properties. This review examines the multifaceted mechanisms that DPs employ to lower lipid levels in subjects with hyperlipidemia. DPs could directly inhibit lipid intake and absorption, promote lipid excretion, and regulate key enzymes involved in lipid metabolism pathways, including triglyceride and cholesterol anabolism and catabolism, fatty acid oxidation, and bile acid synthesis. Additionally, DPs indirectly improve lipid homeostasis by modulating gut microbiota composition and alleviating oxidative stress. Moreover, the lipid-lowering mechanisms of particular structural DPs (including ß-glucan, pectin, glucomannan, inulin, arabinoxylan, and fucoidan) are summarized. The relationship between the structure and lipid-lowering activity of DPs is also discussed based on current researches. Finally, potential breakthroughs and future directions in the development of DPs in lipid-lowering activity are discussed. The paper could provide a reference for further exploring the mechanism of DPs for lipid regulations and utilizing DPs as lipid-lowering dietary ingredients.

A review: Effects of microbial fermentation on the structure and bioactivity of polysaccharides in plant-based foods.

Fermented plant-based foods that catering to consumers' diverse dietary preferences play an important role in promoting human health. Recent exploration of their nutritional value has sparked increasing interest in the structural and bioactive changes of polysaccharides during fermentation, the essential components of plant-based foods which have been extensively studied for their structures and functional properties. Based on the latest key findings, this review summarized the dominant fermented plant-based foods in the market, the involved microbes and plant polysaccharides, and the corresponding modification in polysaccharides structure. Further microbial utilization of these polysaccharides, influencing factors, and the potential contributions of altered structure to the functions of polysaccharides were collectively illustrated. Moreover, future research trend was proposed, focusing on the directional modification of polysaccharides and exploration of the mechanisms underlying structural changes and enhanced biological activity during fermentation.

In vitro digestion and fermentation characteristics of eight kinds of pulses and suggestions for different populations.

Pulse-based diets are attracting attention for their potential in combating diet-related non-communicable diseases. However, limited research studies have focused on the digestive and fermentative properties of pulses, which are crucial for exerting benefits. Here, we investigated the in vitro digestibility of starch/protein, along with the fermentation characteristics, of eight pulses and their pastes, including white kidney beans, adzuki beans, cowpeas, broad beans, mung beans, chickpeas, white lentils, and yellow peas. The findings indicated that pulse flours and pastes were low GL food (estimated GL < 10) and had a low degree of protein hydrolysis during simulated gastrointestinal digestion. During in vitro fermentation, pulses flours and pastes decreased the fermentation pH, increased the level of short-chain fatty acids (mainly consisting of valeric acid, followed by acetic acid, propionic acid, butyric acid, isobutyric acid, and isovaleric acid), and positively modulated the microbiota composition over time, specifically reducing the ratio of Firmicutes to Bacteroidetes. In addition, we found that boiling could affect the in vitro digestion and fermentation characteristics of pulses, possibly depending on their intrinsic nutrient characteristics. This research could provide a comprehensive summary of the nutrient content, digestibility, and fermentation of eight pulses and their pastes. Guided by factor analysis, for different individuals' consumption, pulses, cowpeas, broad beans, white lentils, and white kidney beans were preferred for diabetic individuals, yellow peas and white lentils were preferred for intestinal homeostasis disorders, and white lentils, broad beans, white kidney beans, and cowpeas were suitable for obese individuals, in which white lentils were considered healthier and suggested for healthy adults.

Insoluble/soluble fraction ratio determines effects of dietary fiber on gut microbiota and serum metabolites in healthy mice.

Both soluble dietary fiber (SDF) and insoluble dietary fiber (IDF) play pivotal roles in maintaining gut microbiota homeostasis; whether the effects of the different ratios of IDF and SDF are consistent remains unclear. Consequently, we selected SDFs and IDFs from six representative foods (apple, celery, kale, black fungus, oats, and soybeans) and formulated nine dietary fiber recipes composed of IDF and SDF with a ratio from 1 : 9 to 9 : 1 (NDFR) to compare their impact on microbial effects with healthy mice. We discovered that NDFR treatment decreased the abundance of Proteobacteria and the ratio of Firmicutes/Bacteroidetes at the phylum level. The α diversity and relative richness of Parabacteroides and Prevotella at the genus level showed an upward trend along with the ratio of IDF increasing, while the relative abundance of Akkermansia at the genus level and the production of acetic acid and propionic acid exhibited an increased trend along with the ratio of SDF increasing. The relative abundance of Parabacteroides and Prevotella in the I9S1DF group (the ratio of IDF and SDF was 9 : 1) was 1.72 times and 5.92 times higher than that in the I1S9DF group (the ratio of IDF and SDF was 1 : 9), respectively. The relative abundance of Akkermansia in the I1S9DF group was 17.18 times higher than that in the I9S1DF group. Moreover, a high ratio of SDF (SDF reaches 60% or more) enriched the glycerophospholipid metabolism pathway; however, a high ratio of IDF (IDF reaches 80% or more) regulated the tricarboxylic acid cycle. These findings are helpful in the development of dietary fiber supplements based on gut microbiota and metabolites.

Gut microbial utilization of xylan and its implication in gut homeostasis and metabolic response.

Xylan as the second most abundant indigestible carbohydrate found in nature attracts great interests of researchers, nutritionist and consumers due to its various health benefits. However, accumulated studies indicate the interactions with gut microbiota greatly affect these benefits, and significant progress has been made over the past few years to understand how microbes utilize xylan at gene level. In this review, we focused on gut xylanolytic microbes and xylan's physico-chemical features, summarized the xylanases needed for complete xylan decomposition, their substrate specificity and the presence in gut microbes, as well as microbial degradation of xylan in single strain mode and cooperation mode. Xylan utilization system were discussed with different phyla. Furthermore, the implications on intestinal homeostasis and metabolic response were reviewed with clinical effects emphasized, and highlight is placed on specific gut microbes and the complexity of xylan structure to provide a clue for the inconsistent results in human studies. CHEMICAL COMPOUNDS: xylan; arabinoxylan, glucuronoxylans; glucuronoarabinoxylans; xylo-oligosaccharides; arabinoxylo-oligosaccharides.

Short-term exposure to high relative humidity increases blood urea and influences colonic urea-nitrogen metabolism by altering the gut microbiota.

Introduction: Colonic urea-nitrogen metabolites have been implicated in the pathogenesis of certain diseases which can be affected by environmental factors. Objectives: We aimed to explore the influence of ambient humidity on colonic urea-nitrogen metabolism. Methods: Blood biochemical indexes, metabolites of intestinal tract, and gut microbiota composition of mice (n = 10/group) exposed to high relative humidity (RH, 90 ± 2%) were analyzed during the 14-day exposure. Results: After 12-h exposure, plasma blood urea nitrogen (BUN) level increased along with a decrease in the activity of erythrocyte Na+/K+ -ATPase. Moreover, abnormal erythrocyte morphologies appeared after 3 days of exposure. The colonic BUN and ammonia levels increased significantly after the 12-h and 24-h exposure, respectively. The colonic level of amino acids, partly synthesized by gut microbiota using ammonia as the nitrogen source, was significantly higher on the 7th day. Furthermore, the level of fecal short-chain fatty acids was significantly higher after 3-day exposure and the level of branched-chain fatty acids increased on the 14th day. Overall, gut microbiota composition was continuously altered during exposure, facilitating the preferential proliferation of urea-nitrogen metabolism bacteria. Conclusion: Our findings suggest that short-term high RH exposure influences colonic urea-nitrogen metabolism by increasing the influx of colonic urea and altering gut microbiota, which might further impact the host health outcomes.

Polysaccharides from fermented coix seed modulates circulating nitrogen and immune function by altering gut microbiota.

Coix lachryma-jobi L. seed is an important food item in Asia with culinary and medicinal values. The effects of non-fermented coix seed (NFC), fermented coix seed with Lactobacillus plantarum NCU137 (FC) and polysaccharides from NFC, FC (FCP) on mice circulating nitrogen and immune disorder induced by high relative humidity (RH, 90 ± 2%) exposure were compared. All the treatments reduced circulating nitrogen (BUN and ammonia) might via increasing excretion of fecal nitrogen induced by altering gut microbiota. In comparison, FC and FCP restored erythrocyte morphology by promoting erythrocyte Na+/K+ ATPase activity more effectively, and immune function was modulated by reducing plasma IgM and IFN-γ levels, up-regulating IL-4 and IL-6 levels. Herein, these results indicated that FCP, as the main active ingredient in FC, modulated circulating nitrogen through altering gut microbiota, and restored immune homeostasis by regulating Th1/Th2 cytokines in mice receiving high RH exposure.

Effects of tea polysaccharides in combination with polyphenols on dextran sodium sulfate-induced colitis in mice.

Both tea polysaccharides (TPS) and tea polyphenols (TPP) are promising in the treatment of inflammatory bowel disease (IBD). However, the effects of their combination against IBD are still unknown. In the present study, the therapeutic effects of TPS, TPP and TPS + TPP on dextran sodium sulfate-induced colitis in mice were investigated. Our results showed that administration of TPS + TPP achieved the best effects, followed by TPP and TPS, which were evidenced by the restoration of various physical signs (body weight, colon length and disease activity index) and the promoted intestinal barrier function (colon damage, mucin secretion and tight junction proteins expression). Furthermore, TPP and TPS decreased the relative abundance of Proteobacteria and Enterobacteriaceae, while TPP + TPS increased that of Lactobacillaceae and Lactobacillus. In conclusion, TPS together with TPP had greater effects on alleviating colitis and promoting intestinal barrier function. This result is interesting when developing functional foods against colitis.

In vitro digestion of eight types of wholegrains and their dietary recommendations for different populations.

Wholegrains have been promoted for human consumption due to their various health benefits. However, different wholegrains vary in nutritional composition and their beneficial impact on health. In this study, we compared the in vitro starch and protein digestibility, as well as dietary fiber content of eight different wholegrains including barley, buckwheat, coix seed, foxtail millet, oat, proso millet, quinoa, and sorghum and their porridges. We found that boiling improved starch digestibility of all grains, and protein digestibility except proso millet and sorghum. Porridges made from oats, quinoa, or buckwheat are considered healthier than others due to their lower glycemic index and glycemic load, higher digestible protein content and amino acid bioaccessibility, and higher dietary fiber content (>12%). This study could provide a comprehensive nutritional composition and digestibility of the eight types of wholegrains and their porridges. Dietary recommendations were also given for different populations based on factor analysis.

Microbiota-related effects of prebiotic fibres in lipopolysaccharide-induced endotoxemic mice: short chain fatty acid production and gut commensal translocation.

Fructans such as fructo-oligosaccharides (FOS) and inulin have been reported to directly regulate ileal inflammatory responses in lipopolysaccharide (LPS)-induced endotoxemic mice, without alterations in the colonic microbiota. Firstly, we replicated this model and found that a single gavage of 10 mg g-1 of fructans directly promoted caecal acetate and propionate production. Thus, the previous understanding of microbiota-independent effects of prebiotic fructans in endotoxemic mice has been challenged. In parallel, we performed a daily gavage of 160 mg kg-1 of inulin, xylan, or Dendrobium officinale polysaccharides (DOP) for two weeks prior to LPS injection. The long-term intake of prebiotic fibres reduced the bacterial load in the spleen and mesenteric lymph nodes (MLNs), and in comparison, a single gavage of fructans increased that. However, the long-term intake was unable to improve the short-chain fatty acid (SCFA) synthesis and epithelial barrier function that were impaired by LPS. Notably, the three fibre types consistently reduced the expression of mucin 2 (MUC2) and variously modulated critical mediators (IL-18, IL-22, and HIF-1α) to regulate the host-commensal microbiota interactions in the ileum. In addition, the three fibre types consistently inhibited the inflammatory T helper (Th) cell response in the ileum, while they diversely modulated the peripheral and systemic Th cell responses. Overall, the prebiotic fibres displayed microbiota-related changes in endotoxemic mice, and the potential associations with the in vivo anti-inflammatory effects of prebiotic fibres need further investigation.

Effects of processing parameters on furan formation in canned strawberry jam.

The formation mechanism of furan has been studied extensively in model systems, however, furan formation in real foods are complex and far from being fully understood. In this study, the effects of acid-regulating agent (citric acid), sugar addition (glucose, fructose and sucrose) and thickening agents (xanthan gum, κ-carrageenan and pectin) on furan levels in strawberry jams were studied; meanwhile the formation pathway of furan in canned strawberry jam was proposed by carbon module labeling (CAMOLA) technique. Our results suggested low pH promoted furan formation in strawberry jam. Besides, fructose produces more furans than sucrose and glucose, and the addition of xanthan gum reduced furan levels significantly. The kinetic data showed that ascorbic and dehydroascorbic acid degradation followed first-order kinetics while rate of furan formation followed zero-order kinetics. This study presented the possibility of mitigating furan formation in canned strawberry jams by optimization of processing parameters and addition of xanthan gum.

Effects of fermentation with Lactobacillus plantarum NCU137 on nutritional, sensory and stability properties of Coix (Coix lachryma-jobi L.) seed.

This study aimed to investigate the effect of fermentation with Lactobacillus plantarum NCU137 on the nutritional, sensory and stability properties of Coix (Coix lachryma-jobi L.) seed. The nutritional compounds, including free amino acid, free fatty acid, soluble dietary fiber and organic acids of fermented coix seed were significantly (p < 0.05) increased than those of non-fermented coix seed. The fermented coix seed exhibiting a special flavor, due to the production of acids, the decreased level of aldehydes and ketones, and the increased level of alcohols in the volatile compounds, whereas the amount of hazardous substance 2-pentylfuran was reduced and natural antiseptic hexanoic acid was produced. The increased viscosity together with the larger particle size and the reduced absolute ζ potential contribute to the stability of the fermented coix seed paste system. Therefore, fermentation with L. plantarum NCU137 could improve the nutritional, sensory and stability properties of coix seed.

Effects of insoluble and soluble fibers isolated from barley on blood glucose, serum lipids, liver function and caecal short-chain fatty acids in type 2 diabetic and normal rats.

Large prospective cohort studies suggested an important role of cereal insoluble fiber in the prevention and management of type 2 diabetes, which challenge the traditional view that viscosity and solubility are the main driving factors for these beneficial effects of dietary fiber. To evaluate the anti-diabetic effects of insoluble- (BIF) and soluble fibers (BSF) enzymatically isolated from barley, a conventional rat model and a type 2 diabetes rat model were used. Our results showed that 4-week treatment of BIF or BSF effectively reduced FBG in the diabetic condition, with caecal level of propionic acid and minor SCFAs increased by BIF and that of butyric acid and insulin sensitivity improved by BSF, respectively. The two treatments further ameliorated liver function, judged by the recovered serum level of ALT, albumin and total protein levels. BIF and BSF also increased HDL-C and decreased serum MDA. In normal rats, BIF and BSF showed a hypolipidaemic effect in triglycerides and LDL-C, reduced body weight and enhanced the caecal production of minor SCFAs. Furthermore, the two treatments reduced the caecal level of butyric acid while BSF increased that of propionic acid. In conclusion, BIF could exert anti-diabetic effects that might via a different mechanism from BSF.

pH and lipid unsaturation impact the formation of acrylamide and 5-hydroxymethylfurfural in model system at frying temperature.

The effect of unsaturated free fatty acids (FFAs) and triglycerides (TGs) on the formation of acrylamide and 5-hydroxymethylfurfural (HMF) at frying temperature was investigated. We hypothesized that the nature of the precursors generated by the Maillard reaction in the presence of FFAs and TGs, such as the Amadori product and α-dicarbonyl compounds, would impact acrylamide and HMF formation. Interestingly, the initial pH of the aqueous phase impairs the observed effect of the addition of lipids on the formation of acrylamide. Addition of FFAs and TGs increased acrylamide formation in non-buffered systems (distilled water, pH 4.6), but decreased it in buffered systems (sodium acetate, pH 6.0). Conversely, incorporation of these lipids promoted HMF formation under both conditions. The degree of fatty acid unsaturation played an important role in the formation of acrylamide and HMF. The effect of unsaturated lipids on the acrylamide and HMF was due to lipid oxidation. Statistical correlations were made between acrylamide and HMF formation and α-dicarbonyl compounds in the presence of unsaturated lipids.

Effects of barley variety, dietary fiber and ß-glucan content on bile acid composition in cecum of rats fed low- and high-fat diets.

Diet-induced obesity and insulin resistance have been linked to changes in bile acid (BA) profiles, which in turn are highly dependent on the dietary composition and activity of the gut microbiota. The objective of the present study was to investigate whether the type and level of fiber had an effect on cecal BA composition when included in low- and high-fat diets. Groups of rats were fed two barley varieties, which resulted in three test diets containing three levels of ß-glucans and two levels of dietary fiber. BAs were preconcentrated using hollow fiber liquid-phase microextraction and quantified by gas chromatography. The amount of the secondary BAs, lithocholic-, deoxycholic- and hyodexycholic acids was generally higher in groups fed high-fat diets compared with corresponding acids in groups fed low-fat diets (P<.05). In contrast, most of the primary and the secondary BAs, ursodeoxycholic acid and ß- and ω-muricholic acids, were two to five times higher (P<.05) in groups fed low-fat diets than in groups fed high-fat diets. This was particularly true for groups fed the highest level of ß-glucans and in some cases also the medium level. The BA profile in the gut was strongly dependent on the amount and type of dietary fiber in the diet, which may be useful in the prevention/treatment of diseases associated with changes in BA profiles.

Modulation of gut microbiota in rats fed high-fat diets by processing whole-grain barley to barley malt.

SCOPE: The gut microbiota is linked with human health, and by manipulating its composition, health conditions might be improved. The aim of this study was to investigate whether two barley products, whole-grain barley and barley malt, caused differentiation of the cecal microbiota in rats fed high-fat diets and whether there were correlations with the short-chain fatty acids formed. METHODS AND RESULTS: Male Wistar rats were given barley or malt (7-8 dietary fiber/100 g) for 4 weeks. Cellulose was used as a control, and the cecal microbiota was analyzed with next-generation sequencing of 16S rDNA. The barley group had higher abundances of Verrucomicrobia and Actinobacteria and lower abundances of Firmicutes and Deferribacteres than the control group; the alpha diversity was also lower. At the genus level, the barley group had higher abundances of Akkermansia, Ruminococcus, Blautia, and Bilophila. Turicibacter and Roseburia were more abundant in the malt group, and Parabacteroides, Dorea and rc4-4 were enriched in the control group. Most genera correlated with acetic and propionic acids, but Roseburia and Turicibacter instead correlated with butyric acid. Succinic acid correlated with Clostridium and Akkermansia. CONCLUSION: Bioprocessing is a potential method to modulate the gut microbiota for enhanced effects on human health.

Barley malt increases hindgut and portal butyric acid, modulates gene expression of gut tight junction proteins and Toll-like receptors in rats fed high-fat diets, but high advanced glycation end-products partially attenuate the effects.

Barley malt, a product of controlled germination, has been shown to produce high levels of butyric acid in the cecum and portal serum of rats and may therefore have anti-inflammatory effects. The aim of the study was to investigate how four barley malts, caramelized and colored malts, 50-malt and 350-malt, differing in functional characteristics concerning beta-glucan content and color, affect short-chain fatty acids (SCFA), barrier function and inflammation in the hindgut of rats fed high-fat diets. Male Wistar rats were given malt-supplemented high-fat diets for four weeks. Low and high-fat diets containing microcrystalline cellulose were incorporated as controls. All diets contained 70 g kg(-1) dietary fiber. The malt-fed groups were found to have had induced higher amounts of butyric and propionic acids in the hindgut and portal serum compared with controls, while cecal succinic acid only increased to a small extent. Fat increased the mRNA expression of tight junction proteins and Toll-like receptors (TLR) in the small intestine and distal colon of the rats, as well as the concentration of some amino acids in the portal plasma, but malt seemed to counteract these adverse effects to some extent. However, the high content of advanced glycation end-products (AGE) in caramelized malt tended to prohibit the positive effects on occludin in the small intestine and plasma amino acids seen with the other malt products. In conclusion, malting seems to be an interesting process for producing foods with positive health effects, but part of these effects may be destroyed if the malt contains a high content of AGE.

Prebiotic and synbiotic effects on rats fed malted barley with selected bacteria strains.

BACKGROUND: Butyric acid, one of the key products formed when ß-glucans are degraded by the microbiota in the colon, has been proposed to be important for colonic health. Glutamine bound to the fibre may have similar effects once it has been liberated from the fibre in the colon. Both ß-glucans and glutamine are found in high amounts in malted barley. Lactobacillus rhamnosus together with malt has been shown to increase the formation of butyric acid further in rats. OBJECTIVE: To investigate whether Lactobacillus rhamnosus 271, Lactobacillus paracasei 87002, Lactobacillus plantarum HEAL 9 and 19, and Bifidobacterium infantis CURE 21 affect the levels of short-chain fatty acids and glutamine in caecum and portal blood of rats fed barley malt. DESIGN: The experimental diets were fed for 12 days. The daily dose of the probiotic strain was 1×10(9) colony forming units and the intake of fibre 0.82 g/day. RESULTS: The malt mostly contained insoluble fibre polymers (93%), consisting of glucose and xylose (38-41 g/kg) and some arabinose (21 g/kg). The fibre polysaccharides were quite resistant to fermentation in the rats, regardless of whether or not probiotics were added (25-30% were fermented). Caecal and portal levels of acetic acid decreased in the rats after the addition of L. plantarum HEAL 9 and L. rhamnosus 271, and also the levels of butyric acid. Viable counts of Lactobacillus, Bifidobacterium and Enterobacteriaceae were unaffected, while the caecal composition of Lactobacilli was influenced by the type of strain administrated. Portal levels of glutamine were unchanged, but glycine levels increased with L. plantarum HEAL 9 and 19 and phenylalanine with L. rhamnosus 271. CONCLUSIONS: Although the probiotic strains survived and reached the caecum, except B. infantis CURE 21, there were no effects on viable counts or in the fermentation of different fibre components, but the formation of some bacterial metabolites decreased. This may be due to the high proportion of insoluble fibres in the malt.