Influence of Different Types, Utilization Times, and Volumes of Aging Barrels on the Metabolite Profile of Red Wine Revealed by 1H-NMR Metabolomics Approach.

It is well recognized that the aging process is a critical step in winemaking because it induces substantial chemical changes linked to the organoleptic properties and stability of the finished wines. Therefore, this study aimed to investigate the influence of different types, utilization times, and volumes of aging barrels on the metabolite profile of red wines, produced from Thai-grown Shiraz grapes, using a non-targeted proton nuclear magnetic resonance (1H-NMR) metabolomics approach. As a result, 37 non-volatile polar metabolites including alcohols, amino acids, organic acids, carbohydrates and low-molecular-weight phenolics were identified. Chemometric analysis allowed the discrimination of wine metabolite profiles associated with different types of aging containers (oak barrels vs. stainless-steel tanks), as well as the utilization times (2, 6 and >10 years old) and volumes (225, 500 and 2000 L) of the wooden barrels employed. Significant variations in the concentration of formate, fumarate, pyruvate, succinate, citrate, gallate, acetate, tyrosine, phenylalanine, histidine, γ-aminobutyrate, methionine and choline were statistically suggested as indicators accountable for the discrimination of samples aged under different conditions. These feature biomarkers could be applied to manipulate the use of aging containers to achieve the desired wine maturation profiles.

Synbiotic edible film from konjac glucomannan composed of Lactobacillus casei-01® and Orafti®GR, and its application as coating on bread buns.

BACKGROUND: Konjac glucomannan-based edible films formulated with Lactobacillus casei-01® and chicory-derived inulin Orafti®GR were studied for their properties, stability, and application as coatings on bread buns. RESULTS: Thickness and transparency were variable and dependent on the formulations. Alterations in color properties of all supplemented films were unnoticeable by unaided human eyes, with ΔE less than 3. Lactobacillus casei-01® and Orafti®GR were associated with higher water solubility of the films. Lactobacillus casei-01® decreased the water vapor permeability of the films while Orafti®GR promoted it. The mechanical properties in all combinations remained unchanged, although those with Orafti®GR showed profoundly reduced tensile strength. Scanning electron micrographs and Fourier transform infrared spectra of the films confirmed good homogeneity and intermolecular attraction between the prebiotic and konjac glucomannan. Cell viability in the films stored at room temperature decreased sharply, becoming less than the minimum recommended level after day 4, while viable L. casei-01® in coatings on bread buns gradually decreased, with a reduction of ca. 2 log colony-forming units (CFU) portion-1 over the 7 day storage period at room temperature. CONCLUSION: The synbiotic film and coating developed in this study are a relatively simple strategy for incorporating L. casei-01® and Orafti®GR into bread buns, which are short shelf-life foods. Bread buns with synbiotic coating could diversify functional food choices. Pretreatment, together with other technologies, is required to maintain a desirable number of active probiotic cells for longer. © 2020 Society of Chemical Industry.

Influence of Lactobacillus plantarum WCFS1 on post-acidification, metabolite formation and survival of starter bacteria in set-yoghurt.

The objectives of this study were to evaluate the growth and survival of the model probiotic strain Lactobacillus plantarum WCFS1 in co-culture with traditional yoghurt starters and to investigate the impact of preculturing on their survival and metabolite formation in set-yoghurt. L. plantarum WCFS1 was precultured under sublethal stress conditions (combinations of elevated NaCl and low pH) in a batch fermentor before inoculation in milk. Adaptive responses of L. plantarum WCFS1 were evaluated by monitoring bacterial population dynamics, milk acidification and changes in volatile and non-volatile metabolite profiles of set-yoghurt. The results demonstrated that sublethal preculturing did not significantly affect survival of L. plantarum WCFS1. On the other hand, incorporation of sublethally precultured L. plantarum WCFS1 significantly impaired the survival of Lactobacillus delbrueckii subsp. bulgaricus which consequently reduced the post-acidification of yoghurt during refrigerated storage. A complementary metabolomics approach using headspace SPME-GC/MS and (1)H NMR combined with multivariate statistical analysis revealed substantial impact of sublethally precultured L. plantarum WCFS1 on the metabolite profiles of set-yoghurt. This study provides insight in the technological implications of non-dairy model probiotic strain L. plantarum WCFS1, such as its good stability in fermented milk and the inhibitory effect on post-acidification.

Effect of sublethal preculturing on the survival of probiotics and metabolite formation in set-yoghurt.

The objective of this study was to investigate the effect of preculturing of Lactobacillus rhamnosus GG and Bifidobacterium animalis subsp. lactis BB12 under sublethal stress conditions on their survival and metabolite formation in set-yoghurt. Prior to co-cultivation with yoghurt starters in milk, the two probiotic strains were precultured under sublethal stress conditions (combinations of elevated NaCl and low pH) in a batch fermentor. The activity of sublethally precultured probiotics was evaluated during fermentation and refrigerated storage by monitoring bacterial population dynamics, milk acidification and changes in volatile and non-volatile metabolite profiles of set-yoghurt. The results demonstrated adaptive stress responses of the two probiotic strains resulting in their viability improvement without adverse influence on milk acidification. A complementary metabolomic approach using SPME-GC/MS and (1)H-NMR resulted in the identification of 35 volatiles and 43 non-volatile polar metabolites, respectively. Principal component analysis revealed substantial impact of the activity of sublethally precultured probiotics on metabolite formation demonstrated by distinctive volatile and non-volatile metabolite profiles of set-yoghurt. Changes in relative abundance of various aroma compounds suggest that incorporation of stress-adapted probiotics considerably influences the organoleptic quality of product. This study provides new information on the application of stress-adapted probiotics in an actual food-carrier environment.

Changes in the metabolomic profiles of mammary secretion in relation to dam litter size and parity number in Black Bengal goats.

Background and Aim: The colostrum is essential for a kid's survival and development. The metabolomic profiles of mammary secretion in goats are limited. This study investigated the metabolomic profiles of mammary secretion in purebred Black Bengal goats and their relationships with litter size and parity number. Materials and Methods: 500 MHz nuclear magnetic resonance was used to analyze the metabolomic profiles of 43 colostrum and milk samples collected on delivery day and day 7 after parturition, respectively. Results: Fifty-one metabolites were distinguished between colostrum and milk based on heatmap visualization and hierarchical cluster analysis. In colostrum, most compounds were present in significantly greater amounts than in milk. Milk of goats with multiple litter sizes had higher levels of lactose while fat, protein, total solids, solid not fat, and most of the metabolites were lower. The parity number of dams shows no difference in the composition of all components between primiparous and multiparous goats. Conclusion: The components in colostrum were significantly more concentrated than those in milk. The multiple litter sizes in dams led to a significant impact on the composition of lactose and other milk metabolites.

Physicochemical, Nutritional, and Antioxidant Properties of Traditionally Fermented Thai Vegetables: A Promising Functional Plant-Based Food.

Fermented plant-based products were gathered from various regions in Thailand and categorized into 10 types of traditional commercial vegetables. Different vegetable materials and natural fermentation methods influence the diverse physical, chemical, nutritional, and functional attributes of the products. All the traditionally fermented Thai vegetable samples collected showed physicochemical properties associated with the fermentation process, contributing to the nutritional and functional quality of the final products. Achieving consistent research results is challenging due to the intricate nature of food matrices and biochemical processes during fermentation. The roles of microorganisms, especially probiotics, are crucial in delivering health benefits through fermented foods. Traditionally fermented Thai vegetable foods contain high levels of total soluble solids, titratable acidity, and salinity in pickled shallot and ginger as a result of the natural fermentation process and the ingredients used. The research findings were confirmed using a hierarchical cluster analysis (HCA)-derived dendrogram pattern. The nutritional compositions, total phenolic contents, and antioxidant activities varied among the different types of vegetables. The correlations among lipid, protein, fiber, total soluble solid (TSSs), total titratable acidity (TTA), and salinity as potential biomarkers in fermented vegetable products were examined. The results suggest that traditionally fermented Thai vegetable products significantly impacted food research by enhancing the quality and preserving the authenticity of traditionally fermented Thai vegetables.

Clostridium butyricum Probiotic Feed Additive: Modulation of Sow Milk Metabolomics and Mitigation of Pre-Weaning Piglet Diarrhea.

The present study aimed to investigate the impact of Clostridium butyricum probiotic feed additive on sow and piglet performances, together with alterations in the lipidomic and metabolomic profiles of sow milk. Sixty-four Landrace × Yorkshire crossbred sows and 794 piglets were included. Sows were divided into two groups; i.e., (i) conventional gestation diet (control; n = 35) and (ii) conventional diet added with 10 g/sow/day of probiotic C. butyricum spores (treatment; n = 29) from one week before the estimated farrowing day until weaning (29.6 ± 4.8 days). The sow and piglet performances and incidence of piglet diarrhea were recorded. Changes in gross chemical composition, fatty acid and non-volatile polar metabolite profiles of sow colostrum, transient milk and mature milk were evaluated. The results showed that relative backfat loss in the treatment group (-2.3%) was significantly lower than in control group (11.6%), especially in primiparous sows (p = 0.019). The application of C. butyricum probiotics in sows significantly reduced the incidence of diarrhea in piglets (p < 0.001) but no other effect on piglet performance was found. Lipidomic and metabolomic analyses revealed variations in sow colostrum and milk biomolecular profiles, with indicative compounds significantly altered by feeding with the C. butyricum probiotics. In conclusion, the use of C. butyricum probiotics in sows may improve sow body condition and reduce diarrhea incidence in piglets, with underlying changes in milk composition that warrant further investigation. These findings support the potential of C. butyricum as a beneficial feed additive in swine production.

Factors affecting bacterial community dynamics and volatile metabolite profiles of Thai traditional salt fermented fish.

Bacterial diversity of the Thai traditional salt fermented fish with roasted rice bran (Pla-ra) was investigated using classical and molecular approaches. Bacterial population of Pla-ra ranged from 102-106 in solid-state (SSF) and 106-109 CFU/g in submerged (SMF) fermentation types. Halanaerobium spp. and Lentibacillus spp. were the main genera particularly detected when rRNA analysis was applied. Tetragenococcus halophillus were dominant during the final stage in sea salt-recipe samples while Bacillus spp. were found in those rock salt recipes. In contrast, cultural plating demonstrated that Bacillus spp., generally B. amyloliquefaciens, were the dominant genera. In addition, B. pumilus, B. autrophaeus, B.subtilis and B. velezensis shown some relations with rock salt-recipe samples. The main volatile metabolites in all samples were butanoic acid and its derivatives. Key factors affected bacterial profiles and volatile compounds of salt fermented fish were type of salt, addition of roasted rice bran, and fermenting conditions.

Lacticaseibacilli attenuated fecal dysbiosis and metabolome changes in Candida-administered bilateral nephrectomy mice.

The impacts of metabolomic changes (reduced short-chain-fatty acids; SCFAs) in uremic condition is not fully understood. Once daily Candida gavage with or without probiotics (different times of administration) for 1 week prior to bilateral nephrectomy (Bil Nep) in 8-week-old C57BL6 mice as the possible models more resemble human conditions were performed. Candida-administered Bil Nep mice demonstrated more severe conditions than Bil Nep alone as indicated by mortality (n = 10/group) and other 48 h parameters (n = 6-8/group), including serum cytokines, leaky gut (FITC-dextran assay, endotoxemia, serum beta-glucan, and loss of Zona-occludens-1), and dysbiosis (increased Enterobacteriaceae with decreased diversity in microbiome analysis) (n = 3/group for fecal microbiome) without the difference in uremia (serum creatinine). With nuclear magnetic resonance metabolome analysis (n = 3-5/group), Bil Nep reduced fecal butyric (and propionic) acid and blood 3-hydroxy butyrate compared with sham and Candida-Bil Nep altered metabolomic patterns compared with Bil Nep alone. Then, Lacticaseibacillus rhamnosus dfa1 (SCFA-producing Lacticaseibacilli) (n = 8/group) attenuated the model severity (mortality, leaky gut, serum cytokines, and increased fecal butyrate) of Bil Nep mice (n = 6/group) (regardless of Candida). In enterocytes (Caco-2 cells), butyrate attenuated injury induced by indoxyl sulfate (a gut-derived uremic toxin) as indicated by transepithelial electrical resistance, supernatant IL-8, NFκB expression, and cell energy status (mitochondria and glycolysis activities by extracellular flux analysis). In conclusion, the reduced butyrate by uremia was not enhanced by Candida administration; however, the presence of Candida in the gut induced a leaky gut that was attenuated by SCFA-producing probiotics. Our data support the use of probiotics in uremia.

Dynamics of fatty acid and non-volatile polar metabolite profiles in colostrum and milk depending on the lactation stage and parity number of sows.

The objective of this study was to investigate the impact of lactation stage and parity number on fatty acid and non-volatile polar metabolite profiles in sow colostrum and milk using a metabolomics approach. A total number of 63 colostrum, transient and mature milk were collected from primiparous and multiparous Landrace × Yorkshire crossbred sows. Macrochemical, fatty acid and non-volatile polar metabolite compositions of samples were analyzed using infrared spectrometry, gas chromatography coupled with mass-spectrometry and proton nuclear magnetic resonance spectroscopy, respectively. Univariate and multivariate statistical analysis demonstrated significant impacts of lactation stage and parity number on colostrum and milk compositions. Chemometric analysis revealed significant influences of sow parity on the distinction in fatty acid profiles of mature milk while the distinction in non-volatile polar metabolite profiles was more evident in colostrum. Alterations in the concentration of linoleic (C18:2n6), lignoceric (C24:0), behenic (C22:0), caprylic (C8:0) and myristoleic (C14:1) acid together with those of creatine, creatinine phosphate, glutamate and glycolate were statistically suggested to be mainly affected by sow parity number. Variations in the concentration of these compounds reflected the physiological function of sow mammary gland influenced. This information could be applied for feed and feeding strategies in lactating sows and improving lactating performances.

Arthrospira platensis Mutagenesis for Protein and C-Phycocyanin Improvement and Proteomics Approaches.

Arthrospira (Spirulina) platensis is known for its use as a food supplement, with reported therapeutic properties including antiviral, anti-inflammatory and antioxidant activity. Arthrospira is also an excellent source of proteins and C-phycocyanin. The latter is a light-harvesting pigment-protein complex in cyanobacteria, located on the outer surface of the thylakoid membrane and comprising 40 to 60% of the total soluble protein in cells. Random mutagenesis is a useful tool as a non-genetically modified mutation method that has been widely used to generate mutants of different microorganisms. Exposure of microalgae or cyanobacteria to chemical stimuli affects their growth and many biological processes. Chemicals influence several proteins, including those involved in carbohydrate and energy metabolisms, photosynthesis and stress-related proteins (oxidative stress-reactive oxygen species (ROS) scavenging enzymes). Signal transduction pathways and ion transportation mechanisms are also impacted by chemical treatment, with changes causing the production of numerous biomolecules and stimulation of defence responses. This study compared the protein contents of A. platensis control and after mutagenesis using diethyl sulphate (DES) under various treatment concentrations for effective mutation of A. platensis. Results identified 1152 peptides using proteomics approaches. The proteins were classified into 23 functional categories. Random mutagenesis of A. platensis by DES was found to be highly effective for C-phycocyanin and protein production.

Lactobacillus acidophilus LA5 improves saturated fat-induced obesity mouse model through the enhanced intestinal Akkermansia muciniphila.

Obesity, a major healthcare problem worldwide, induces metabolic endotoxemia through the gut translocation of lipopolysaccharides (LPS), a major cell wall component of Gram-negative bacteria, causing a chronic inflammatory state. A combination of several probiotics including Lactobacillus acidophilus 5 (LA5), a potent lactic acid-producing bacterium, has previously been shown to attenuate obesity. However, data on the correlation between a single administration of LA5 versus microbiota alteration might be helpful for the probiotic adjustment. LA5 was administered daily together with a high-fat diet (HFD) for 8 weeks in mice. Furthermore, the condition media of LA5 was also tested in a hepatocyte cell-line (HepG2 cells). Accordingly, LA5 attenuated obesity in mice as demonstrated by weight reduction, regional fat accumulation, lipidemia, liver injury (liver weight, lipid compositions, and liver enzyme), gut permeability defect, endotoxemia, and serum cytokines. Unsurprisingly, LA5 improved these parameters and acidified fecal pH leads to the attenuation of fecal dysbiosis. The fecal microbiome analysis in obese mice with or without LA5 indicated; (i) decreased Bacteroidetes (Gram-negative anaerobes that predominate in non-healthy conditions), (ii) reduced total fecal Gram-negative bacterial burdens (the sources of gut LPS), (iii) enhanced Firmicutes (Gram-positive bacteria with potential benefits) and (iv) increased Verrucomycobia, especially Akkermansia muciniphila, a bacterium with the anti-obesity property. With LA5 administration, A. muciniphila in the colon were more than 2,000 folds higher than the regular diet mice as determined by 16S rRNA. Besides, LA5 produced anti-inflammatory molecules with a similar molecular weight to LPS that reduced cytokine production in LPS-activated HepG2 cells. In conclusion, LA5 attenuated obesity through (i) gut dysbiosis attenuation, partly through the promotion of A. muciniphila (probiotics with the difficulty in preparation processes), (ii) reduced endotoxemia, and (iii) possibly decreased liver injury by producing the anti-inflammatory molecules.

Influence of different proteolytic strains of Streptococcus thermophilus in co-culture with Lactobacillus delbrueckii subsp. bulgaricus on the metabolite profile of set-yoghurt.

Proto-cooperation between Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus is one of the key factors that determine the fermentation process and final quality of yoghurt. In this study, the interaction between different proteolytic strains of S. thermophilus and L. delbrueckii subsp. bulgaricus was investigated in terms of microbial growth, acidification and changes in the biochemical composition of milk during set-yoghurt fermentation. A complementary metabolomics approach was applied for global characterization of volatile and non-volatile polar metabolite profiles of yoghurt associated with proteolytic activity of the individual strains in the starter cultures. The results demonstrated that only non-proteolytic S. thermophilus (Prt-) strain performed proto-cooperation with L. delbrueckii subsp. bulgaricus. The proto-cooperation resulted in significant higher populations of the two species, faster milk acidification, significant abundance of aroma volatiles and non-volatile metabolites desirable for a good organoleptic quality of yoghurt. Headspace SPME-GC/MS and (1)H NMR resulted in the identification of 35 volatiles and 43 non-volatile polar metabolites, respectively. Furthermore, multivariate statistical analysis allows discriminating set-yoghurts fermented by different types of starter cultures according to their metabolite profiles. Our finding underlines that selection of suitable strain combinations in yoghurt starters is important for achieving the best technological performance regarding the quality of product.