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AIMS Use of nonsteroidal anti-inflammatory drugs (NSAIDs) can cause damage to the gastric and duodenal mucosa. Some probiotics have proven useful in ameliorating the harmful side-effects of NSAIDs. Our aim was to evaluate whether oral administration of Bifidobacterium animalis ssp. lactis 420 (B420) can attenuate the increase of calprotectin excretion into faeces induced by intake of diclofenac sustained-release tablets. METHODS A double-blind, parallel-group, placebo-controlled and randomized clinical study was performed in 50 healthy male and female volunteers aged 20-40 years, in Finland. Study participation consisted of 4 phases: run-in, intervention with B420 or placebo, B420 or placebo + NSAID treatment, and follow-up. The primary outcome was the concentration of calprotectin in faeces. Secondary outcomes were haemoglobin and microbial DNA in faeces and blood haemoglobin levels. RESULTS Intake of diclofenac increased the faecal excretion of calprotectin in both groups. The observed increases were 48.19 ± 61.55 μg/g faeces (mean ± standard deviation) in the B420 group and 31.30 ± 39.56 μg/g in the placebo group (difference estimate 16.90; 95% confidence interval: -14.00, 47.77; P = .276). There were no significant differences between the treatment groups in changes of faecal or blood haemoglobin. Faecal B. lactis DNA was much more abundant in the B420 group compared to the placebo group (ANOVA estimate for treatment difference 0.85 × 109 /g faeces; 95% confidence interval: 0.50 × 109 , 1.21 × 109 ; P

Polydextrose (PDX) is a branched glucose polymer, utilized as a soluble dietary fiber. Recently, PDX was found to have hypolipidemic effects and effects on the gut microbiota. To investigate these findings more closely, a non-targeted metabolomics approach, was exploited to determine metabolic alterations in blood and epididymal adipose tissue samples that were collected from C57BL/6 mice fed with a Western diet, with or without oral administration of PDX. Metabolomic analyses revealed significant differences between PDX- and control mice, which could be due to differences in diet or due to altered microbial metabolism in the gut. Some metabolites were found in both plasma and adipose tissue, such as the bile acid derivative deoxycholic acid and the microbiome-derived tryptophan metabolite indoxyl sulfate, both of which increased by PDX. Additionally, PDX increased the levels of glycine betaine and l-carnitine in plasma samples, which correlated negatively with plasma TG and positively correlated with bacterial genera enriched in PDX mice. The results demonstrated that PDX caused differential metabolite patterns in blood and adipose tissues and that one-carbon metabolism, associated with glycine betaine and l-carnitine, and bile acid and tryptophan metabolism are associated with the hypolipidemic effects observed in mice that were given PDX.

The effects of orally consumed probiotics on skin wrinkles and dryness are not fully known. A randomized, placebo-controlled, triple-blinded study was conducted with 148 healthy Korean female volunteers aged between 33 and 60 years, who were administered 1.75 × 109 colony-forming units (CFUs) of Bifidobacterium animalis subsp. lactis Bl-04 (Bl-04) (N = 74) or matching placebo (N = 74) for 12 weeks. Facial wrinkles (with 3-dimensional (3D) imaging), skin hydration, transepidermal water loss (TEWL), elasticity, and gloss were assessed at baseline and after every 4 weeks of the intervention. Questionnaire-based subjective evaluations of product efficacy and usability were also analyzed. The consumption of Bl-04 was safe and ameliorated significantly facial skin wrinkle parameters (total wrinkle area and volume, average depth of wrinkles, and arithmetic average roughness (Ra)) versus placebo at 4 weeks, but there were no differences at Week 8 or 12 between groups. Skin hydration, TEWL, elasticity, and gloss were similar between treatment groups, as were the subjective evaluation scores. Oral consumption of Bl-04 indicated promising short-term effects on skin appearance from the winter toward the spring. In future study designs, special attention should be paid to environmental conditions as well as to the skin condition and age of the participants.

There is increasing evidence for a gut-skin axis and a potential influence of the skin microbiota on skin health. With this in mind, there is an opportunity for probiotics, even though they have traditionally been investigated for their intestinal and immune benefits. The skin has an important function as a barrier. Here, oral and topical probiotics, parabiotics, and postbiotics have been observed to have positive benefit, also in supporting the skin against UV-induced damage. The manufacturing of probiotics for skin health, especially for topical applications, may have very specific requirements, different from current oral applications, because the intended use and regulation are different. For the future, the field would benefit from rigorous human intervention studies as are becoming more common in the nutritional field. While currently mainly typical probiotic organisms such as lactobacilli and bifidobacteria are used, it can be foreseen that in the future also organisms representative for the skin microbiota will find application.

BackgroundTight junctions (TJs) between keratinocytes are important for regulating the inside-out and outside-in permeability barriers. Betaine is a natural intracellular osmolyte utilized for instance in cosmetics. We determined by liquid chromatography that betaine content in human skin samples was higher in the epidermis than in the dermis. This led us to further investigate the effect of betaine on TJs by utilizing different in vitro models: topical application of betaine on normal human epidermal keratinocytes (NHEKs) differentiated for 5-days, freshly isolated NHEKs differentiated for 3 days and betaine applied during the differentiation process of organotypic rat epidermal keratinocytes (REKs) differentiated for 12-15 days. ResultsTopical betaine in amount range of 100 to 500 µM increased transepithelial electrical resistance (TEER) in 5-day-differentiated NHEKs, and downregulated occludin mRNA in concentrations 100-250 µM, but not claudin-4 (CLDN4) or zonula occludens-1 (ZO-1). In REKs, betaine applied during differentiation process had no effect on mannitol permeation or the expression of occludin, ZO-1, transforming growth factor beta 2 (TGF-β2), or cluster of differentiation (CD147). However, in freshly isolated differentiating human epidermal keratinocytes, betaine treatment increased TJ protein ZO-1 and the differentiation marker involucrin at protein level. By indirect immunofluorescence, prominent staining and co-localization for TJ proteins ZO-1 and claudin-1 was detected in freshly isolated differentiating keratinocytes treated with betaine. ConclusionsTaken together, the results provide evidence that betaine modulates TJ proteins in differentiating epidermal keratinocytes and promotes formation of TJs. These results suggest a role for betaine in the regulation of the epidermal barrier and as a natural ingredient in topical cosmetics.

An increased intake of fruit and vegetables has been associated with decreased risk of cardiovascular disease (CVD). However, it is currently unclear as to whether particular fruits and vegetables convey more benefit than others, and if the form in which a particular fruit or vegetable is delivered, for example, depending on polyphenol content or the degree of processing, affects potential health benefits. The aim of this study was to conduct a pilot randomised placebo-controlled trial (RCT) with the ultimate aim of assessing if different apple products differentially affect CVD risk factors. The RCT was conducted in those at elevated risk of CVD. Participants were randomised to receive high polyphenol apples, low polyphenol apples, capsules containing apple extract, dried apple pieces or placebo capsules to take for four weeks. A range of risk factors and biomarkers associated with CVD were measured before and after the intervention period. A total of 55 participants were randomised and completed the study. Change in hip circumference was significantly different across the groups after 4 weeks (P=0.02) Change in fibre intake was also statistically significant between groups, with those consuming high polyphenol apples having a higher intake than those on the apple extract (P=0.01). There was a significant within-group change in fasting oxidised LDL in the apple extract group (P=0.008). A significant difference in change in volume-corrected epicatechin was observed (P=0.03), with those on apple extract having higher levels than those on placebo capsule (P=0.002) or low polyphenol apples (P=0.01) Overall, interventions were generally acceptable to participants. Apple products appeared to have a positive effect on some risk factors for CVD in this pilot study, which indicated a definitive RCT is feasible. Further adequately powered studies need to be conducted to definitively test these findings.

Introduction: Dietary fibers help to control energy intake and reduce the risk of developing obesity. Recent studies show that the consumption of polydextrose reduces energy intake at a subsequent meal. In this systematic review and meta-analysis we examine the subsequent effects of polydextrose on different levels of energy intake (EI). Method: The review followed the PRISMA methodology. Meta-analyses were expressed as Standardized Mean Difference (SMD). A linear regression approach was used to model the relationship between the polydextrose dose and the different levels of EI expressed as a relative change (%). Results: All the studies included in this review administered polydextrose as part of a mid-morning snack. Six studies were included in the analysis of EI at an ad libitum lunch; and three were included in the analysis of EI during the rest of the day, as well as total daily EI. The meta-analysis showed that the consumption of polydextrose is associated with a reduction in EI at lunch time (SMD = 0.35; P 0.01; I 2 = 0). The dose of polydextrose consumed correlated significantly with this reduction in EI, EI Lunch (%) = −0.67 Polydextrose (g/day) (R 2 = 0.80; P 0.01). The meta-analysis of EI during the rest of the day and daily EI did not show any difference. Nevertheless, the regression equation indicates that there is a dose-dependent effect on the reduction of daily EI, EI Daily (%) = −0.35 × Polydextrose (g/day) (R 2 = 0.68; P 0.05). Sex-specific results are consistent with results for the whole group. Conclusion: The studies included in this meta-analysis support the notion that the consumption of polydextrose reduces voluntary energy intake at a subsequent meal. Furthermore, this reduction in energy intake occurs in a dose-dependent manner.

Scope High-fat diets are a likely cause of low-grade inflammation and obesity-related pathologies. This study measures the effects of a high-fat diet, in combination with two dietary supplements-betaine and polydextrose-on metabolism and inflammation in the adipose tissue of diet-induced obese mice. Methods and results Forty male C57BL/6J mice are fed a high-fat diet for 8 weeks and compared with low-fat-diet-fed control animals (n = 10). For the last 4 weeks, the high-fat-diet-fed animals are supplemented with 1% betaine, 3.33% polydextrose, their combination, or plain water. Fat depots from subcutaneous and visceral adipose tissue are analyzed for inflammatory markers and nontargeted metabolomics by quantitative PCR and LC-QTOF-MS. The high-fat diet significantly increases adipose tissue inflammation in both fat depots. By metabolic profiling, clear differences are noted between low-fat-diet and high-fat-diet groups with regard to the levels of several metabolite species-primarily carnitines, lipids, and amino acids. Dietary betaine mitigates the high-fat-diet-induced IL-6 expression and significantly increases betaine and butyrobetaine levels in adipose tissue. Conclusions The high-fat diet induces patent changes in carnitine and lipid metabolism in adipose tissue. Betaine supplementation elevates the levels of betaine and its derivatives and certain carnitine species, as reported in muscle and liver, and moderately reduces inflammation.

Xylitol has been widely documented to have dental health benefits, such as reducing the risk for dental caries. Here we report on other health benefits that have been investigated for xylitol. In skin, xylitol has been reported to improve barrier function and suppress the growth of potential skin pathogens. As a non-digestible carbohydrate, xylitol enters the colon where it is fermented by members of the colonic microbiota; species of the genus Anaerostipes have been reported to ferment xylitol and produce butyrate. The most common Lactobacillus and Bifidobacterium species do not appear to be able to grow on xylitol. The non-digestible but fermentable nature of xylitol also contributes to a constipation relieving effect and improved bone mineral density. Xylitol also modulates the immune system, which, together with its antimicrobial activity contribute to a reduced respiratory tract infection, sinusitis, and otitis media risk. As a low caloric sweetener, xylitol may contribute to weight management. It has been suggested that xylitol also increases satiety, but these results are not convincing yet. The benefit of xylitol on metabolic health, in addition to the benefit of the mere replacement of sucrose, remains to be determined in humans. Additional health benefits of xylitol have thus been reported and indicate further opportunities but need to be confirmed in human studies.

OBJECTIVES: Human breast milk contains a high quantity of oligosaccharides (HMOs) which are not digested by infants but are selectively utilized by beneficial gut microbes such as Bifidobacterium species. The gut microbiome in breast-fed infants is dominated by Bifidobacterium species, whereas formula-fed infants have a higher representation of Bacteroides and Enterobacteria species. The aim of this study was to evaluate the ability of various potentially pathogenic bacteria and commercially available probiotic strains to grow using the HMOs 2’-fucosyllactose (2’-FL) and 3-fucosyllactose (3-FL), glucose, lactose or galacto-oligosaccharide (GOS) as a sole carbon source. We also compared the production of short-chain fatty acids (SCFAs) and lactic acid during the fermentation of 2’-FL by B. infantis, B. bifidum and Bacteroides strains. METHODS: Bacterial growth was monitored by measuring the absorbance at 600 nm every 30 min for 24 h using the automatic Bioscreen(©) C system under anaerobic conditions in culture media containing 1% 2’-FL, 3-FL, glucose, lactose or GOS as a sole carbon source. Metabolites were analyzed by chromatographic methods in spent culture media after over-night growth of bacterial strains with 1% 2’-FL. RESULTS: Only certain bifidobacteria strains, B. infantis and B. bifidum, were able to utilize 2’-FL and 3-FL, whereas other tested Lactobacillus and Bifidobacterium strains as well as pathogenic bacteria grew with all other carbon sources. We also confirmed that Bacteroides fragilis, B. thetaiotaomicron and B. vulgatus strains utilized HMO. Distinct SCFA and lactic acid quantities were observed between Bifidobacterium and Bacteroides strains in 2’-FL fermentation. For instance, acetic and lactic acids were produced at much higher levels in Bifidobacterium than in Bacteroides, and propionic acid was produced at low level only in Bacteroides. CONCLUSIONS: These results show the selectivity of 2’-FL and 3-FL utilization in certain Bifidobacterium and Bacteroides strains, which is translated to differences in metabolite production between these individual HMO-utilizing bacteria. Further studies will enlighten the role and potential benefits of HMOs as a selective substrate for gut microbes, as well as for competition and cross-feeding of microbial metabolites among gut species. FUNDING SOURCES: This research was funded by DuPont Nutrition and Health.

Given that vascular calcification is inversely correlated with the clinical intake of menaquinone, a rat model of warfarin-induced calcification may be useful for testing menaquinone and vitamin K-1 potential effects on vascular function. The aim of the present study was to investigate effects of vitamin K-1 and menaquinone-7 treatments on blood pressure and vascular function in warfarin-induced vascular calcification model during five-week intervention in normotensive Wistar-Kyoto rats. Blood pressure was measured weekly, and at the end of the intervention in vitro vascular reactivity measurements were done. Alizarin Red S and von Kossa stainings were used to record possible calcification of aortic sections. Routine clinical chemistry was done from serum and urine samples. Vascular calcification was seen only in a few warfarin-treated animals in histological staining. Warfarin-treatment did not change significantly blood pressure of the rats. Warfarin-treatment increased slightly the endothelium-dependent relaxation of aorta after the L-type calcium channels were blocked. Also the vascular relaxation improved after NOS inhibition in the aorta of the healthy controls and menaquinone-7 treated animals, indicating that the relaxation in those groups was not totally dependent on NO. Clinical chemistry from serum showed some differences in urea, creatinine as well as lipid and glucose metabolism between the healthy controls and warfarin-treated rats.

Essential oils (EOs) are important aromatic components of herbs and spices and their biological activities have been known and utilised since ancient times in perfumery, food preservation, flavouring, and medi - cine. Some of their biological activities include antibacterial, antifungal, anti-oxidant and anti-inflammatory effects amongst others. EOs have received attention in recent years as potential 'natural' alternatives for replacing antibiotic growth promoters (AGPs) in animal diets due to their positive impact on growth performance, gut microbiota and welfare. The purpose of this paper is to provide an overview of our own published and unpublished data on the antibacterial, antifungal and insecticidal activity of thymol and cinnamaldehyde (TC blend), and to describe the effects of this specific EO blend on gut microbiota, growth performance and welfare, carcass characteristics and food safety. The possible modes of action of EOs are discussed and areas for future research are proposed.

Scope: Betaine (BET) reduces diet-induced liver lipid accumulation, and may relieve obesity-related metabolic disturbances. The aim of our study was to analyze metabolite alterations after supplementation of BET, polydextrose (PDX, a soluble dietary fiber), or their combination (BET PDX) via drinking water to C57BL/6J mice fed a high-fat (HF) diet. Methods and results: BET supplementation increased BET levels in plasma, muscle, and liver (p < 0.05), and the nontargeted LC-MS metabolite profiling revealed an increase in several metabolites in the carnitine biosynthesis pathway after BET supplementation both in liver and muscle. These included carnitine and acetylcarnitine (1.4-fold, p < 0.05), propionylcarnitine and ?-butyrobetaine (1.5-fold, p < 0.05), and several other short-chain acylcarnitines (p < 0.05) in muscle. These changes were slightly higher in the BET PDX group. Furthermore, BET reduced the HF diet induced accumulation of triglycerides in liver (p < 0.05). The supplementations did not attenuate the HF diet induced increase in body weight gain or the increase in adipose tissue mass. Instead, the combination of BET and PDX tended to increase adiposity. Conclusion: Our results suggest that increased availability of BET in different tissues, especially in muscle, after BET supplementation has an impact on carnitine metabolism, and this could further explain the link between BET and lipid metabolism.

Flavanol-rich foods are known to exert beneficial effects on cardiovascular health. The biological effects depend on bioavailability of flavanols which may be influenced by food matrix and dose ingested. We compared the bioavailability and dose-response of epicatechin from whole apple and an epicatechin-rich extract, and the effects on plasma and urinary nitric oxide (NO) metabolites.In a randomized, placebo-controlled, crossover trial, subjects consumed drinks containing 70 and 140 mg epicatechin from an apple extract and an apple puree containing 70 mg epicatechin. Blood and urine samples were collected for 24 h post ingestion. Maximum plasma concentration, AUC(0-24 h) , absorption and urinary excretion were all significantly higher after ingestion of both epicatechin drinks compared with apple puree (p0.05). Time to maximum plasma concentration was significantly later for the puree compared with the drinks (p0.01). Epicatechin bioavailability was2-fold higher after ingestion of the 140 mg epicatechin drink compared to the 70 mg epicatechin drink (p0.05). Excretion of NO metabolites was higher for all test products compared with placebo, which was significant for the high dose drink (p = 0.016).Oral bioavailability of apple epicatechin increases at higher doses, is reduced by whole apple matrix and has the potential to increase NO bioavailability.

The aim was to develop novel fibres by enzymatic synthesis, to determine their total dietary fibre by AOAC method 2009.01 and to estimate their potential digestibility and assess their digestibility in vivo using glycaemic and insulinaemic responses as markers in mice and randomised clinical trial models. We found that fibre candidates to which α-(1,2) branching was added were resistant to digestion in the mouse model, depending on the amount of branching. These results show that in vivo models are needed to reliably assess the digestibility of α-glycosidic-linked oligomeric dietary fibre candidates, possibly due to absence of brush border α-glucosidase activity in the current in vitro assessment. α-(1,3)-linked and α-(1,6)-linked glucose oligomers were completely digested in humans and mice. In conclusion, it is possible to develop dietary soluble fibres by enzymatic synthesis. Adding α-(1,2) branching increases their resistance to digestion in vivo and can thus improve their suitability as potential fibre candidates. Clinical Trial Registry: ClinicalTrials.gov, NCT02701270.

Polydextrose (PDX) reduces subsequent energy intake (EI) when administered at midmorning in single-blind trials of primarily normal-weight men. However, it is unclear if this effect also occurs when PDX is given at breakfast time. Furthermore, for ecological validity, it is desirable to study a female population, including those at risk for obesity. We studied the effects of PDX, served as part of a breakfast or midmorning preload, on subsequent EI and other appetite-related parameters in healthy normal-weight and overweight females. Per earlier studies, the primary outcome was defined as the difference in subsequent EI when PDX was consumed at midmorning versus placebo. Thirty-two volunteers were enrolled in this acute, double-blind, placebo-controlled, randomized, and crossover trial to examine the effects of 12.5 g of PDX, administered as part of a breakfast or midmorning preload, on subsequent EI, subjective feelings of appetite, well-being, and mood. Gastric emptying rates and the blood concentrations of glucose, insulin, cholecystokinin, ghrelin, glucagon-like peptide 1 (GLP-1), and peptide tyrosine-tyrosine were measured in the group that received PDX as part of their breakfast. There were no differences in EI between volunteers who were fed PDX and placebo. PDX intake with breakfast tended to elevate blood glucose (P = 0.06) during the postabsorptive phase, significantly lowered insulin by 15.7% (P = 0.04), and increased GLP-1 by 39.9% (P = 0.02); no other effects on blood parameters or gastric emptying rates were observed. PDX intake at midmorning reduced hunger by 31.4% during the satiation period (P = 0.02); all other subjective feelings of appetite were unaffected. Volunteers had a uniform mood profile during the study. PDX was well tolerated, causing one mild adverse event throughout the trial. © 2016 The Authors

Obesity is related to the consumption of energy-dense foods in addition to changes in the microbiome where a higher abundance of gut Bacteroidetes can be found in lean subjects or after weight loss. Lactitol, a sweet-tasting sugar alcohol, is a common sugar-replacement in foods. Polydextrose, a highly branched glucose polymer, is known to reduce energy intake. Here, we test if the combined effects of lactitol or polydextrose in combination with Bacteroides species will have a beneficial metabolic response in rats fed a high-fat diet. 175 male Wistar rats were fed either a low-fat or high-fat diet. Bacteroides thetaiotaomicron (1010 bacteria/animal/day) was orally administered with or without lactitol (1.6−2 g/animal/day) or polydextrose (2 g/animal/day) for 8 days. Postprandial blood samples, cecal digesta and feces were collected on the last day. Measurements included: body weight, feed consumption, cecal short chain fatty acids, fecal dry matter and heat value, blood glucose, insulin, triglyceride, and satiety hormone concentrations. Lactitol and polydextrose decreased the mean body weight when administered with B. thetaiotaomicron or when lactitol was administered alone. Levels of postprandial plasma triglycerides declined with lactitol and polydextrose when administered with B. thetaiotaomicron. For intestinal hormone release, lactitol − alone or with B. thetaiotaomicron − increased the release of gastrointestinal peptide tyrosine tyrosine (PYY) as well as the area under the curve (AUC) measured for PYY (0−8 h). In addition, levels of insulin AUC (0−8 h) decreased in the lactitol and polydextrose supplemented groups. Lactitol and polydextrose may both provide additional means to regulate postprandial metabolism and weight management whereas the addition of B. thetaiotaomicron in the tested doses had only minor effects on the measured parameters.

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

The microbes in our gut can influence our weight by providing us with energy through the degradation of nondigestable carbohydrates and by affecting the cellular energy status of liver and muscle cells and the accumulation of lipids in adipose tissue. Thus, it is not surprising that in several studies the gastrointestinal microbiota of overweight and obese subjects has been found to differ from that of lean subjects. The initial findings linked obesity with proportionally decreased levels of the phylum Bacteroidetes and increased levels of the phylum Firmicutes. Later, several studies have assessed the association between overweight or obesity and the gastrointestinal microbiota, applying an array of molecular methods targeting the microbiota as a whole or specific bacterial groups or species within. However, at present it is difficult to draw conclusions on which of the observed microbiota alterations are relevant; essentially all of the bacterial groups that have been studied in more than one trial have given contradictory results in regard to their association with weight. Some of these discrepancies can result from methodological issues and some from the nature of the gastrointestinal microbiota, which is an extremely complex and dynamic microbial ecosystem with high subject specificity. In addition, selecting subjects purely based on weight may result in a largely heterogeneous group with several potentially confounding factors. While it may be premature to conclude which specific groups of bacteria are prominent in the intestinal tract of overweight and obese subjects, it appears clear that microbes contribute to weight gain and related health issues, such as the metabolic syndrome and type II diabetes. Therefore, it is important to continue to search for common microbial markers and predictors of obesity, and to study how these may be modulated with probiotics and prebiotics to promote health.

1. In this study the effect of a blend of essential oils (EO) comprising 15 g/tonne thymol and 5 g/tonne cinnamaldehyde on the performance and intestinal microbiota of broilers was investigated. 2. A total of 720 male Ross broilers were divided into two dietary treatments with 12 replicate pens per treatment. Broilers were given a control soybean-wheat-based diet with or without added EO in two diet phases (0-21 d and 22-42 d). 3. The blend of EO increased body weight gain of broilers from 0 to 42 d by 45%. 4. Caecal microbiota were affected by the EO blend; in particular increases in the proportions of Lactobacillus and Escherichia coli at 41 d was observed. 5. The EO blend had major effects on caecal metabolites. The proportion of caecal butyrate at 20 and 41 d of age increased, whereas the proportion of caecal acetic acid at 20 d, and propionic acid and isovaleric acid at 41 d, decreased with the EO blend. In addition, the caecal proportion of spermine increased and tyramine decreased at 41 d of age with the EO treatment. 6. The present study shows that EO supplementation exerts a positive effect on intestinal microbiota with a concomitant enhancement in growth performance. The study suggests that modulation of broiler gut microbiota composition and activity through the administration of EO offers an effective means for improving broiler performance.

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

The use of antimicrobial growth promoters has been banned in the EU. This has created an inter - est in alternative strategies to prevent an imbalance in the intestinal microbiota and the potential development of intestinal disorders in livestock. Essential oils (EOs) have been known to exhibit antimicrobial activity against specific microbial species and could therefore be considered one such alternative in controlling the intestinal micro - bial population. Under anaerobic conditions, the tested Clostridium perfringens strains were found to be sensitive (P < 0.05) to carvacrol, cinnamaldehyde, citral, limonene, thymol, particularly at the higher concentration tested (500 mg/l) and to oregano oil, rosemary oil and thyme oil. Streptococcus epidermis was sensitive (P < 0.05) to most EO's tested, also mainly at the higher concentration. The tested Salmonella serovars were found to be sensitive (P < 0.05) only to high (500 mg/l) concentrations of the tested EOs. Escherichia coli was sensitive (P < 0.05) to most of the tested EOs, also at lower concentrations (5 and 50 mg/l). Bifidobacterium longum, Bifidobacterium breve and Lactobacillus reuteri were less sensitive (P < 0.05) to most of the tested EOs, while Bifidobacterium animalis ssp. lactis and L. fermentum were relatively sensitive also at lower concentrations (5 and 50 mg/l), although growth reduction by EOs of these bacterial species was less then with the antimicrobial growth promoter avilamycin. With the exception of Salmonella and E. coli, all tested microbes were sensitive to avilamycin. Selected EOs seem to have the advantage of inhibiting the growth of potential pathogens while only moderately influencing beneficial members of the intestinal microbiota. This difference in sensitivity may strengthen the microbiota and contribute to improved animal health.

A cross-sectional study was carried out in order to compare intestinal microbiological and immunological biomarkers with blood glucose and lipids, satiety-related hormones and inflammatory biomarkers characterising differences between obese and normal weight subjects. Faecal and blood samples were obtained from twenty obese subjects with an average BMI of 32·9 kg/m2and twenty normal weight subjects with an average BMI of 23·3 kg/m2. Blood insulin, TAG and leptin were significantly elevated, whereas concentrations of HDL and ghrelin were significantly decreased in the obese subjects. Inflammatory status in the obese subjects was characterised by a trend for elevated blood C-reactive protein (CRP;P = 0·06) and IL-6 (P = 0·02). The faecal microbial composition differed between the groups; less sulphate-reducing bacteria (P = 0·05) and a trend for lessBacteroides(P = 0·07) were measured for overweight subjects. Furthermore, an inverse correlation was demonstrated between faecalBacteroideslevels and waist circumference (P = 0·05). The faecal microbial metabolites differed between the groups; increased concentrations of branched-chain fatty acids, phenolics, valeric acid, di- and hydroxy acids were described in the obese subjects. No differences between the measured intestinal inflammatory biomarkers were detected. However, systemic inflammation (CRP and IL-6) was correlated with the faecal concentrations of phenolics and lactic acid (P P

Ageing has been suggested to cause changes in the intestinal microbial community. In the present study, the microbiota of a previously well-defined group of elderly subjects aged between 70 and 85 years, both non-steroidal anti-inflammatory drugs (NSAID) users (n9) and non-users (n9), were further compared with young adults (n14) with a mean age of 28 years, by two DNA-based techniques: percentage guanine+cytosine (%G+C) profiling and 16S rDNA sequencing. Remarkable changes in microbiota were described with both methods: compared with young adults a significant reduction in overall numbers of microbes in both elderly groups was measured. Moreover, the total number of microbes in elderly NSAID users was higher than in elderly without NSAID. In 16S rDNA sequencing, shifts in all major microbial phyla, such as lower numbers of Firmicutes and an increase in numbers of Bacteroidetes in the elderly were monitored. On the genus level an interesting link between reductions in the proportion of known butyrate producers belonging toClostridiumcluster XIVa, such asRoseburiaandRuminococcus, could be demonstrated in the elderly. Moreover, in the Actinobacteria group, lower numbers ofCollinsellaspp. were evident in the elderly subjects with NSAID compared both with young adults and the elderly without NSAID, suggesting that the use of NSAID along with age may also influence the composition of intestinal microbiota. Furthermore, relatively high numbers ofLactobacillusappeared only in the elderly subjects without NSAID. In general, the lowered numbers of microbial members in the major phyla, Firmicutes, together with changes in the epithelial layer functions can have a significant effect on the colon health of the elderly.

Introduction: Subjective feelings of appetite are measured using visual analogue scales (VAS) in controlled trials. However, the methods used to analyze VAS during the Satiation (pre- to post-meal) and Satiety (post-meal to subsequent meal) periods vary broadly, making it difficult to compare results amongst independent studies testing the same product. This review proposes a methodology to analyze VAS during both the Satiation and Satiety periods, allowing us to compare results in a meta-analysis. Methods: A methodology to express VAS results as incremental areas under the curve (iAUC) for both the Satiation and Satiety periods is proposed using polydextrose as a case study. Further, a systematic review and meta-analysis on subjective feelings of appetite was conducted following the PRISMA methodology. Meta-analyses were expressed as Standardized Mean Difference (SMD). Results: Seven studies were included in the meta-analysis. There were important differences in the methods used to analyze appetite ratings amongst these studies. The separate subjective feelings of appetite reported were Hunger, Satisfaction, Fullness, Prospective Food Consumption, and the Desire to Eat. The method proposed here allowed the results of the different studies to be homogenized. The meta-analysis showed that Desire to Eat during the Satiation period favors polydextrose for the reduction of this subjective feeling of appetite (SMD = 0.24, I2 < 0.01, p = 0.018); this effect was also significant in the sub-analysis by sex for the male population (SMD = 0.35, I2 < 0.01, p = 0.015). There were no other significant results. Conclusion: It is possible to compare VAS results from separate studies. The assessment of iAUC for both the Satiation and Satiety periods generates results of homogeneous magnitudes. This case study demonstrates, for the first time, that polydextrose reduces the Desire to Eat during the Satiation period. This may explain, at least in part, the observed effects of polydextrose on the reduction of levels of energy intake at subsequent meals.

Objective: Betaine (trimethylglycine) is a naturally-occurring osmolyte and methyl donor with reported effects on liver metabolism. The aim of this study was to evaluate the safety and the effects of betaine administration to metabolic risk factors in Asian males. Methodology: Twenty Japanese males (30-59 years) with mild fatty liver were enrolled in this blinded, placebocontrolled, randomized, parallel trial to assess the effects of 3 g of daily betaine for 12 weeks on safety and metabolic biomarkers. Selection criteria included fatty liver disease, as assessed by an abdominal ultrasonogram (US), the practice of moderate alcohol consumption, liver performance status and body mass index (≤27). Abdominal computed tomography (CT) as well as blood and urine sampling were performed before and after the intervention period. Results: No differences in the degree of hepatic steatosis or in the occurrence of adverse events were observed between the groups. Compared to the placebo group, the betaine group showed a significant increase in plasma betaine (P

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

The effects of various dietary fibres on gut health have been studied extensively but their combined effects are scarcely documented. In the present study the effects of 2 % (w/w) polydextrose (PDX), 2 % (w/w) disaccharide lactitol, or 2 % (w/w) PDX+2 % (w/w) lactitol on gut microflora, microbial metabolism and gut immune responses were investigated in rats. Both PDX and lactitol alone had an effect on many of the studied parameters, but their combination had stronger than additive effects in some parameters. The PDX+lactitol combination altered the microbial community structure as seen by a culture-independent method, percentage guanine+cytosine (%G+C) profiling, increasing the areas of %G+C 35–39 (PP=0·0002), and decreasing %G+C 65–74 (PP=0·03) and 50 % (P=0·002), respectively, indicating a shift from putrefactive towards saccharolytic metabolism. PDX increased the secretion of IgA in the caecum (P=0·007). Secretion of IgA increased even more, almost ten-fold, with the combination of PDX+lactitol (PP

Betaine and its precursor choline were compared in their efficiency in affecting the performance, carcass traits, and liver betaine concentration of growing-finishing pigs. Individually penned Finnish Landrace and Yorkshire pigs and their crosses (30 kg; no. = 70) were offered the basal diet with no added betaine or choline, or the basal diet supplemented with low to moderate doses (250, 500 or 1000 mg/kg) of betaine (Betafin® S1), or with a similar molar amount of choline (578, 1155 or 2310 mg/kg of choline chloride). The maize-soya-bean-meal basal diet was formulated to contain 12·3 MJ/kg digestible energy, 155 g/kg crude protein and 7·4, 4·4 and 4·3 g/kg digestible lysine, threonine and methionine + cystine, respectively. Oat hull meal (100 g/kg) was added to reduce the dietary energy concentration. The pigs were on a restricted feeding level, 1·5 to 3·0 kg food per day (proportionately 0·8 of ad libitum intake) for 75 days. Daily weight gain and food-to-gain ratio improved linearly (P < 0·01) with increasing dietary betaine. Carcass weight increased linearly (P < 0·01) but slaughter loss proportion, backfat and sidefat thicknesses and lean proportions in ham and carcass were unaffected by dietary betaine level. Liver betaine level increased linearly (by up to a proportion of 0·62 in comparison with the control) with dietary betaine addition (F < 0·05) and betaine tended to improve linearly the tensile strength of the proximal ileum (P = 0·07). The presence of choline had no effect on any of these parameters. These results indicate that low to moderate doses of dietary betaine improved the growth and the efficiency of food utilization of growing-finishing pigs. Pigs on betaine diets had heavier carcasses without a relative increase in carcass fat. Choline had no such effects in pigs offered the restricted amount of diet. Liver betaine concentration increased with level of betaine in the diet whereas the betaine precursor choline did not affect hepatic betaine.

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

The efficiency of betaine absorption into small intestinal slices of broiler chicks was studied in vitro with 14C-labeled betaine. The relative proportion of Na+-coupled betaine uptake, as well as the total uptake capacity was larger in the duodenum than in the jejunum. Dietary betaine increased the Na+-coupled uptake in the duodenum. In in vivo-experiments, methyl-14C-labeled betaine, methionine, or choline was fed to broiler chicks. Betaine appeared in the blood more rapidly, and reached a higher total concentration than choline or methionine. The data suggest that choline and methionine were associated with plasma lipoproteins whereas betaine remained free in the plasma. The label distribution in liver, kidney, and intestinal tissues was studied 24 h after label ingestion. Most of the label from betaine was found in the aquaeous phase in the muscle, while in the liver and jejunum the label from betaine was distributed more evenly between the aquaeous, lipid, and protein phases. Label from choline accumulated in the lipid fraction, particularly so in the liver, whereas label from methionine showed a more variable distribution pattern. The distribution results are interpreted in terms of specific roles of betaine, choline, and methionine in methyl group metabolism.

We investigated the mechanisms of pyruvate transport in the erythrocytes of an ancient marine agnathan, the Pacific hagfish (Eptatretus stouti), and a sedentary euryhaline teleost, the starry flounder (Platichthys stellatus). Uptake of [14C]pyruvate (50 µM, 10°C) by flounder erythrocytes was slow (t1/2(half-life) ~ 30 min), nonconcentrative, and mediated by the band 3 Cl-/HCO-3exchanger in combination with a process similar to the H+/monocarboxylate symporter present in freshwater teleosts and mammalian erythrocytes. In contrast, pyruvate uptake by hagfish erythrocytes (50 µM, 10°C) was rapid (t1/2~ 1.5 min) and, in 10 min, reached an intracellular concentration more than 20-fold higher than that present in the extracellular medium. Pyruvate accounted for almost 90% of the accumulated intracellular radioactivity, the remaining label being incorporated into tricarboxylic acid cycle intermediates and glutamate. Influx of pyruvate was saturable (apparent Km= 12 mM) and inhibited by p-chloromercuriphenylsulphonate (PCMBS) (Ki= 71 µM) and 4,4'-diisothiocyanatostilbene-2,2'-disulphonate (DIDS) (Ki= 0.49 mM). Transport was inhibited poorly by α-cyano-4-hydroxycinnamate (CIN) (Ki> 4 mM) and was not coupled to the movement of protons. Instead, the influx of pyruvate was Na+dependent. A sigmoidal relationship between pyruvate transport and extracellular Na+concentration was observed, suggesting a Na+:pyruvate coupling ratio greater than 1:1. In contrast with previously described Na+-dependent monocarboxylate transport activities in mammalian renal and intestinal epithelia, the hagfish erythrocyte system did not transport lactate.

This chapter characterises betaine and its authorised health claim in respect to dietary sources, scientific substantiation, benefits for consumers, interaction with the EU legislation and safety. The European Commission has authorised a health claim for betaine on the contribution to normal homocysteine metabolism. Betaine is obtained from natural dietary sources and is also allowed in food supplements in the EU, and this is examined from the view of condition of use, e.g. the adequate daily intake of betaine. The aim is to characterise the value of betaine related to benefits and health risks for the general population. A question is raised concerning consumer understanding of the authorised heath claim and especially the word homocysteine.

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

Polydextrose (Litesse®, DuPont) is a polysaccharide that is partially fermented in the colon. Evidence suggests that polydextrose increases satiety when consumed over several weeks; however studies assessing its acute effects on satiety are lacking. This study therefore aimed to assess the impact of different doses of polydextrose on satiety and energy intake at subsequent meals during a test day. Three yogurt-based drinks containing different amounts of polydextrose (0, 6.25 and 12.5g) were tested using a randomised, single-blinded, placebo controlled, cross-over design. Thirty-four healthy male and female volunteers were provided with a standard breakfast, then consumed the test product mid-morning, 90min before an ad libitum lunch, which was followed by an ad libitum dinner. Visual analogue scales were used to measure subjective ratings of appetite, liking and discomfort. Consuming 6.25 and 12.5g polydextrose increased satiety and decreased appetite compared to control immediately after consumption. A reduction in energy intake (218.8kJ) at lunchtime was observed for 12.5g polydextrose. This reduction in energy intake was not compensated for at dinner. This study suggests that polydextrose may aid in increasing satiety feelings post consumption and also reduce energy intake as a result.

Obesity is characterised by a state of chronic low-grade inflammation and the elevated circulating and tissue levels of inflammatory markers, including inflammation-related adipokines, released from white adipose tissue. The expression and release of these adipokines generally rises as the adipose tissue expands and hypoxic conditions start to develop within the tissue. Here, the effect of betaine, a trimethylglycine having a biological role as an osmolyte and a methyl donor, on the expression of inflammation-related markers was tested in human adipocytes under hypoxia. Differentiated adipocytes were cultivated under low (1 %) oxygen tension for 8–20 h. The expression of different adipokines, including IL-6, leptin, PPARγ, TNF-α and adiponectin, was measured by quantitative PCR by determining the relative mRNA level from the adipocytes. Hypoxia, in general, led to a decrease in the expression of PPARγ mRNA in human adipocytes, whereas the expression levels of leptin and IL-6 mRNA were substantially increased by hypoxia. The cultivation of adipocytes under hypoxia also led to a reduction in the expression of TNF-α mRNA. The results showed that hypoxia increased the relative quantification of leptin gene transcription, and that betaine (250 μmol/l) reduced this effect, caused by low oxygen conditions. Under hypoxia, betaine also reduced the mRNA level of the pro-inflammatory markers IL-6 and TNF-α. These results demonstrate that the extensive changes in the expression of inflammation-related adipokines in human adipocytes caused by hypoxia can be diminished by the presence of physiologically relevant concentrations of betaine.

In contrast to mammalian erythrocytes which fulfil their energy requirements via anaerobic glycolysis, fish erythrocytes obtain most of their energy aerobically via the Krebs cycle. This enables them to use various substrates, including monocarboxylic acids, glucose and amino acids. This review discusses the membrane permeability of these substrates, and their relative importance in the energy production of fish erythrocytes. Agnathans are characterized by a high permeability to all of the potential substrates, glucose, monocarboxylic and amino acids. In contrast, teleost erythrocytes are often characterized by low glucose permeability. It appears that monocarboxylic acids and certain amino acids such as glutamine may be more important in energy production of teleost erythrocytes than glucose.

Gastrointestinal (GI) adverse effects such as erosion and increased permeability are common during the use of nonsteroidal anti-inflammatory drugs (NSAIDs). Our objective was to assess whetherBifidobacterium animalisssp.lactis420 protects against NSAID-induced GI side effects in a rat model. A total of 120 male Wistar rats were allocated into groups designated as control, NSAID, and probiotic. The NSAID and probiotic groups were challenged with indomethacin (10 mg/kg−1; single dose). The probiotic group was also supplemented daily with 1010 CFU ofB. lactis420 for seven days prior to the indomethacin administration. The control group rats received no indomethacin or probiotic. The permeability of the rat intestine was analysed using carbohydrate probes and the visual damage of the rat stomach mucosa was graded according to severity.B. lactis420 significantly reduced the indomethacin-induced increase in stomach permeability. However, the protective effect on the visual mucosal damage was not significant. The incidence of severe NSAID-induced lesions was, nevertheless, reduced from 50% to 33% with the probiotic treatment. To conclude, theB. lactis420 supplementation protected the rats from an NSAID-induced increase in stomach permeability and may reduce the formation of more serious GI mucosal damage and/or enhance the recovery rate of the stomach mucosa.

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

Non-starch polysaccharides (NSPs) are an important part of our diet and play an essential role in the development of functional foods. NSP are a diverse group of carbohydrates consisting primarily of components of plant wall cells, including gums, plant mucilages, algal polysaccharides and other polysaccharides such as inulin and polydextrose. The current chapter describes the physiological effects of NSP; in most cases as prebiotics. Furthermore, NSPs give structure, mouthfeel and bulk to foods, which are relevant properties especially for the formulation of functional foods, which may be foods with reduced fat and/or sugar.