Repeated Oral Exposure to N ε-Carboxymethyllysine, a Maillard Reaction Product, Alleviates Gut Microbiota Dysbiosis in Colitic Mice.

BACKGROUND: Diet is suggested to participate in the etiology of inflammatory bowel diseases (IBD). Repeated exposure to Maillard reaction products (MRPs), molecules resulting from reduction reactions between amino acids and sugars during food heating, has been reported to be either potentially detrimental or beneficial to health. AIMS: The aim of this study is to determine the effect of repeated oral ingestion of N ε-carboxymethyllysine (CML), an advanced MRP, on the onset of two models of experimental IBD and on the gut microbiota composition of mice. METHODS: Mice received either saline (control) or N ε-carboxymethyllysine daily for 21 days. For the last week of treatment, each group was split into subgroups, receiving dextran sulfate sodium salt (DSS) or trinitrobenzenesulfonic acid (TNBS) to induce colitis. Intensity of inflammation was quantified, and cecal microbiota characterized by bacterial 16S ribosomal RNA (rRNA) amplicon sequencing. RESULTS: Daily oral administration of N ε-carboxymethyllysine did not induce intestinal inflammation and had limited impact on gut microbiota composition (Bacteroidaceae increase, Lachnospiraceae decrease). DSS and TNBS administration resulted in expected moderate experimental colitis with a shift of Bacteroidetes/Firmicutes ratio and a significant Proteobacteria increase but with distinct profiles: different Proteobacteria taxa for DSS, but mainly Enterobacteriaceae for TNBS. While N ε-carboxymethyllysine exposure failed to prevent the inflammatory response, it allowed maintenance of healthy gut microbiota profiles in mice treated with DSS (but not TNBS). CONCLUSIONS: Repeated oral exposure to CML limits dysbiosis in experimental colitis. IBD patients may modulate their microbiota profile by regulating the level and type of dietary MRP consumption.

Microorganisms and Maillard reaction products: a review of the literature and recent findings.

Research on the impact of Maillard reaction products (MRPs) on microorganisms has been reported in the literature for the last 60 years. In the current study, the impact of an MRP-rich medium on the growth of three strains of Escherichia coli was measured by comparing two classic methods for studying the growth of bacteria (plate counting and optical density at 600 nm) and by tracing MRP utilisation. Early stage and advanced MRPs in the culture media were assessed by quantifying furosine and N (ε) -carboxymethyllysine (CML) levels, respectively, using chromatographic methods. These measures were performed prior to and during bacterial growth to estimate the potential use of these MRPs by Escherichia coli CIP 54.8. Glucose and lysine, the two MRP precursors used in the MRP-rich medium, were also quantified by chromatographic means. Compared to control media, increased lag phases and decreased growth rates were observed in the MRP-rich medium for two out of the three Escherichia coli strains tested. In contrast, one strain isolated from the faeces of a piglet fed on a MRP-rich diet was not influenced by the presence of MRPs in the medium. Overall, CML as well as the products obtained by the thermal degradation of glucose and lysine, regardless of the Maillard reaction, did not affect the growth of the three strains tested. In addition, no degradation of fructoselysine or CML was found in the presence of Escherichia coli CIP 54.8.

Ozone Treatments for Preserving Fresh Vegetables Quality: A Critical Review.

Ozone is recognized as an antimicrobial agent for vegetables storage, washing, and processing. This strong disinfectant is now being used in the food industry. In this review, the chemical and physical properties of ozone, its generation, and factors affecting ozone processing efficiency were explained as well as recent regulatory developments in the food industry. By then selecting three vegetables, we show that ozone avoids and controls biological growth on vegetables, keeping their attractive appearance and sensorial qualities, assuring nutritional characteristics' retention and maintaining and increasing the shelf-life. In liquid solution, ozone can be used to disinfect processing water and vegetables, and in gaseous form, ozone helps to sanitize and preserve vegetables during storage. The multifunctionality of ozone makes it a promising food processing agent. However, if ozone is improperly used, it causes some deleterious effects on products, such as losses in their sensory quality. For an effective and a safe use of ozone, specific treatment conditions should be determined for all kinds of vegetables. In a last step, we propose highlighting the different essential characteristics of ozone treatment in order to internationally harmonize the data relating to the treatments carried-out.

Gut Microbiota Modulation by Dietary Barley Malt Melanoidins.

Melanoidins are the final Maillard reaction products (protein-carbohydrate complexes) produced in food by prolonged and intense heating. We assessed the impact of the consumption of melanoidins from barley malts on gut microbiota. Seventy-five mice were assigned into five groups, where the control group consumed a non-melanoidin malt diet, and other groups received melanoidin-rich malts in increments of 25% up to 100% melanoidin malts. Feces were sampled at days 0, 1, 2, 3, 7, 14, and 21 and the microbiota was determined using V4 bacterial 16S rRNA amplicon sequencing and short-chain fatty acids (SCFA) by gas chromatography. Increased melanoidins was found to result in significantly divergent gut microbiota profiles and supported sustained SCFA production. The relative abundance of Dorea, Oscillibacter, and Alisitpes were decreased, while Lactobacillus, Parasutterella, Akkermansia, Bifidobacterium, and Barnesiella increased. Bifidobacterium spp. and Akkermansia spp. were significantly increased in mice consuming the highest melanoidin amounts, suggesting remarkable prebiotic potential.

Effects of Maillard Reaction Products on Sensory and Nutritional Qualities of the Traditional French Baguette.

The goals of this study were to evaluate the effect of baking time on the Maillard reaction products (MRPs) generated in the crust of the traditional French baguette and to estimate their impact on its sensory characteristics, its acrylamide content, and its bifidogenic effect. Melanoidins, volatile compounds, and acrylamide were evaluated in the crust of traditional French baguettes baked between 12 and 22 min at 225 °C. The increase in melanoidins was positively correlated to the baking time, while volatile compounds only increased until 18 min. The acrylamide content was estimated to be below 18 µg/kg, which confirms the findings of EFSA that bread is not a main contributor to dietary acrylamide. A descriptive sensory analysis showed that the baking time positively affected the sensory quality of the crust. The consumer test revealed the same trend and the panelists favorably judged the well-baked baguettes based on a better crust flavor and crispness. The bifidogenic effect of the crust and the crumb from the baguettes baked 22 min was evaluated on the in vitro growth of Bifidobacterium adolescentis. The results demonstrated that the crumb and the crust had exactly the same bifidogenic impact, therefore not caused by melanoidins. PRACTICAL APPLICATION: The consumption of bread in France has decreased since 2007, although bread is considered by French people as "healthy" and essential to maintain a balanced diet. The current study evaluated the global qualities of the French baguette in order to highlight its high sensory quality and its beneficial effect by inducing a possible growth of bifidobacteria, even in well-baked baguettes. These findings allow the French bakery industry to develop an argument to promote its technical know-how and to help consumers choose healthier and tastier bread. Moreover, this study provided some recommendations of baking processes to maintain a high sensory quality of the French baguette and limit the formation of undesirable compounds, such as acrylamide, in the crust.

Polyphenol Interactions Mitigate the Immunogenicity and Allergenicity of Gliadins.

Wheat allergy is an IgE-mediated disorder. Polyphenols, which are known to interact with certain proteins, could be used to reduce allergic reactions. This study screened several polyphenol sources for their ability to interact with gliadins, mask epitopes, and affect basophil degranulation. Polyphenol extracts from artichoke leaves, cranberries, apples, and green tea leaves were examined. Of these extracts, the first three formed insoluble complexes with gliadins. Only the cranberry and apple extracts masked epitopes in dot blot assays using anti-gliadin IgG and IgE antibodies from patients with wheat allergies. The cranberry and artichoke extracts limited cellular degranulation by reducing mouse anti-gliadin IgE recognition. In conclusion, the cranberry extract is the most effective polyphenol source at reducing the immunogenicity and allergenicity of wheat gliadins.

Characterisation of Early-Life Fecal Microbiota in Susceptible and Healthy Pigs to Post-Weaning Diarrhoea.

Early-life microbial exposure is of particular importance to growth, immune system development and long-lasting health. Hence, early microbiota composition is a promising predictive biomarker for health and disease but still remains poorly characterized in regards to susceptibility to diarrhoea. In the present study, we aimed to assess if gut bacterial community diversity and composition during the suckling period were associated with differences in susceptibility of pigs to post-weaning diarrhoea. Twenty piglets from 5 sows (4 piglets / litter) were weaned in poor housing conditions to challenge their susceptibility to post-weaning diarrhoea. Two weeks after weaning, 13 pigs exhibited liquid faeces during 2 or 3 days and were defined as diarrhoeic (D) pigs. The other 7 pigs did not have diarrhea during the whole post-weaning experimental periodand were defined as healthy (H) pigs. Using a molecular characterisation of fecal microbiota with CE-SSCP fingerprint, Next Generation Sequencing and qPCR, we show that D and H pigs were mainly discriminated as early as postnatal day (PND) 7, i.e. 4 weeks before post-weaning diarrhoea occurence. At PND 7 H pigs displayed a lower evenness and a higher abundance of Prevotellaceae, Lachnospiraceae, Ruminocacaceae and Lactobacillaceae compared to D pigs. The sPLS regression method indicates that these bacterial families were strongly correlated to a higher Bacteroidetes abundance observed in PND 30 H pigs one week before diarrhoea. These results emphasize the potential of early microbiota diversity and composition as being an indicator of susceptibility to post-weaning diarrhoea. Furthermore, they support the health promoting strategies of pig herds through gut microbiota engineering.

The fluorimetric FAST method, a simple tool for the optimization of microwave pasteurization of milk.

The effect of microwave pasteurization of cow's milk on its nutritional quality was examined by the FAST method (Fluorescence of Advanced Maillard products and Soluble Tryptophan). Raw milk samples were submitted to different laboratory scale in batch microwave treatments using a central composite experimental design based on specific power and treatment time. The FAST index and bacterial count were monitored to assess protein denaturation, modification by the Maillard reaction and pasteurization efficiency, respectively. High discrimination between samples indicated that the FAST method is a potent tool for estimating the deterioration of the milk quality during experimental microwave treatment. Thus, the FAST index can be effectively used as the continuous response in experimental designs set up and to maximize information economically. In short, the FAST method allows us to retain the rapidity of experimental design while providing the advantages of convenience and low cost.

Maillard reaction products in bread: A novel semi-quantitative method for evaluating melanoidins in bread.

The aim of this study was to test the methods currently in use and to develop a new protocol for the evaluation of melanoidins in bread. Markers of the early and advanced stages of the Maillard reaction were also followed in the crumb and the crust of bread throughout baking, and in a crust model system. The crumb of the bread contained N(ε)-fructoselysine and N(ε)-carboxymethyllysine but at levels 7 and 5 times lower than the crust, respectively. 5-Hydroxymethylfurfural was detected only in the crust and its model system. The available methods for the semi-quantification of melanoidins were found to be unsuitable for their analysis in bread. Our new method based on size exclusion chromatography and fluorescence measures soluble fluorescent melanoidins in bread. These melanoidin macromolecules (1.7-5.6 kDa) were detected intact in both crust and model system. They appear to contribute to the dietary fibre in bread.

The impact of raw materials and baking conditions on Maillard reaction products, thiamine, folate, phytic acid and minerals in white bread.

The aim of this study was to develop a white bread with improved nutrient contents and reduced levels of potentially harmful Maillard reaction products such as N(ε)-carboxymethyllysine (CML) and 5-hydroxymethylfurfural (HMF). Assays were carried out through a full factorial experimental design allowing the simultaneous analysis of four factors at two levels: (1) wheat flour extraction rates (ash content: 0.60%-0.72%), (2) leavening agents (bakers' yeast - bakers' yeast and sourdough), (3) prebaking and (4) baking conditions (different sets of time and temperature). The baking conditions affected HMF and CML as well as certain mineral contents. A reduced baking temperature along with a prolonged heat treatment was found to be favourable for reducing both the CML (up to 20%) and HMF concentrations (up to 96%). The presence of sourdough decreased the formation of CML (up to 28%), and increased the apparent amounts of calcium (up to 8%) and manganese (up to 17.5%) probably through acidification of the dough. The extraction rate of flours as well as interactions between multiple factors also affected certain mineral content. However, compounds like folate, thiamine, copper, zinc, iron and phytic acid were not affected by any of the factors studied.

Insights into bread melanoidins: fate in the upper digestive tract and impact on the gut microbiota using in vitro systems.

Bread melanoidins are heterogeneous, nitrogen-containing, brown macromolecules generated during the last stages of the Maillard reaction in bread. The aim of this study was to investigate the impact and fate of these bread melanoidins in the human gastrointestinal tract using in vitro systems. Batch systems as well as the TNO gastrointestinal tract were used for studying the digestion of various bread samples. These samples included bread crumb, bread crust and two bread-crust-simulating models: a fiber-free model (gluten, starch and glucose heated together) and its control, free of Maillard reaction products (gluten heated separately than starch and glucose). Furthermore, the impact of these two bread-crust-simulating models on the gut microbiota was assessed using a static anaerobic batch system. Bread melanoidins from bread crust and its model were shown to be partially digested by amylases and proteases, suggesting that these melanoidins have peptidic as well as glycosidic bonds in their skeleton. The impact of bread melanoidins from the bread-crust-simulating models and their digestion products on the gut microbiota revealed an individual-dependent response for most flora except for enterobacteria. This flora decreased by -22%, -48% & -100% depending on the individual. Thus, bread melanoidins seem to exert an anti-inflammatory effect by inhibiting enterobacteria.

A laboratory protocol for determining glucose and maximum ethanol production from wheat grain: application to a complete genetic set of near-isogenic waxy lines.

A laboratory protocol was developed to assess glucose and ethanol yields from wheat. The impact of the analyzed wholemeal flour quantity and the saccharification on the amount of released glucose was estimated. The whole process including the analytical methods (glucose and ethanol) was repeatable and reproducible. This protocol was used to assess the glucose and ethanol yields of six varieties and of a complete set of hexaploid near-isogenic waxy lines of cv. Trémie grown in three locations. As compared to the normal line of Trémie, double null (AnBnD) and triple null (nAnBnD) isogenic lines showed a low Hagberg falling number (218, 65, and 63 s, respectively), a higher grain protein content (10.7, 11.5, and 12.1% DM, respectively), a lower glucose yield (728, 703, and 707 kg/t, respectively), and a lower ethanol yield (463, 453, and 452 L/t, respectively). These values indicate a strong involvement of alleles encoded at Wx-B1 and Wx-D1 loci in grain composition.

Juvenile ferric iron prevents microbiota dysbiosis and colitis in adult rodents.

AIM: To assess whether juvenile chronic ferric iron ingestion limit colitis and dysbiosis at adulthood in rats and mice. METHODS: Two sets of experiments were designed. In the first set, recently weaned mice were either orally administered ferrous (Fe²âº) iron salt or ferric (Fe³âº) microencapsulated iron for 6 wk. The last week of experiments trinitrobenzene sulfonic acid (TNBS) colitis was induced. In the second set, juvenile rats received the microencapsulated ferric iron for 6 wk and were also submitted to TNBS colitis during the last week of experiments. In both sets of experiments, animals were sacrificed 7 d after TNBS instillation. Severity of the inflammation was assessed by scoring macroscopic lesions and quantifying colonic myeloperoxidase (MPO) activity. Alteration of the microflora profile was estimated using quantitative polymerase chain reaction (qPCR) by measuring the evolution of total caecal microflora, Bacteroidetes, Firmicutes and enterobacteria. RESULTS: Neither ferrous nor ferric iron daily exposures at the juvenile period result in any effect in control animals at adulthood although ferrous iron repeated administration in infancy limited weight gain. Ferrous iron was unable to limit the experimental colitis (1.71 ± 0.27 MPO U/mg protein vs 2.47 ± 0.22 MPO U/mg protein in colitic mice). In contrast, ferric iron significantly prevented the increase of MPO activity (1.64 ± 0.14 MPO U/mg protein) in TNBS-induced colitis. Moreover, this positive effect was observed at both the doses of ferric iron used (75 and 150 mg/kg per day po--6 wk). In the study we also compared, in both rats and mice, the consequences of chronic repeated low level exposure to ferric iron (75 mg/kg per day po--6 wk) on TNBS-induced colitis and its related dysbiosis. We confirmed that ferric iron limited the TNBS-induced increase of MPO activity in both the rodent species. Furthermore, we assessed the ferric iron incidence on TNBS-induced intestinal microbiota dysbiosis. At first, we needed to optimize the isolation and quantify DNA copy numbers using standard curves to perform by qPCR this interspecies comparison. Using this approach, we determined that total microflora was similar in control rats and mice and was mainly composed of Firmicutes and Bacteroidetes at a ratio of 10/1. Ferric juvenile administration did not modify the microflora profile in control animals. Total microflora numbers remained unchanged whichever experimental conditions studied. Following TNBS-induced colitis, the Firmicutes/Bacteroidetes ratio was altered resulting in a decrease of the Firmicutes numbers and an increase of the Bacteroidetes numbers typical of a gut inflammatory reaction. In parallel, the subdominant population, the enterobacteria was also increased. However, ferric iron supplementation for the juvenile period prevented the increase of Bacteroidetes and of enterobacteria numbers consecutive to the colitis in both the studied species at adulthood. CONCLUSION: Rats and mice juvenile chronic ferric iron ingestion prevents colitis and dysbiosis at adulthood as assessed by the first interspecies comparison.