Preliminary characterization of gut mycobiome enterotypes reveals the correlation trends between host metabolic parameter and diet: a case study in the Thai Cohort.

The association between the gut mycobiome and its potential influence on host metabolism in the Thai Cohort was assessed. Two distinct predominant enterotypes, Saccharomyces (Sa) and Aspergillus/Penicillium (Ap/Pe) showed differences in gut mycobiota diversity and composition. Notably, the Sa enterotype exhibited lower evenness and richness, likely due to the prevalence of Saccharomyces, while both enterotypes displayed unique metabolic behaviors related to nutrient metabolism and body composition. Fiber consumption was positively correlated with adverse body composition and fasting glucose levels in individuals with the Sa enterotype, whereas in the Ap/Pe enterotype it was positively correlated with fat and protein intake. The metabolic functional analysis revealed the Sa enterotype associated with carbohydrate metabolism, while the Ap/Pe enterotype involved in lipid metabolism. Very interestingly, the genes involved in the pentose and glucuronate interconversion pathway, such as polygalacturonase and L-arabinose-isomerase, were enriched in the Sa enterotype signifying a metabolic capacity for complex carbohydrate degradation and utilization of less common sugars as energy sources. These findings highlight the interplay between gut mycobiome composition, dietary habits, and metabolic outcomes within the Thai cohort studies.

Synergistic activity of Limosilactobacillus reuteri KUB-AC5 and water-based plants against Salmonella challenge in a human in vitro gut model.

A synbiotic is a combination of live microorganisms and specific substrates that are selectively utilized by host microorganisms, resulting in health benefits for the host. Previous studies have demonstrated the protective effects of L. reuteri KUB-AC5 against Salmonella infection in chicken and mouse models. The probiotic activity of L. reuteri KUB-AC5 in these hosts was influenced by nutritional supplements. Water-based plants contain significant amounts of carbohydrates, particularly dietary fiber and proteins, making them potential prebiotic substrates. In this study, four water-based plants (Ulva rigida, Caulerpa lentillifera, Wolffia globosa, and Gracillaria fisheri) were screened for their ability to support the growth of L. reuteri KUB-AC5. Under monoculture testing, U. rigida exhibited the highest capacity to support the growth of L. reuteri KUB-AC5 and the production of organic acids, including acetic acid, lactic acid, and propionic acid (p ≤ 0.05). In co-culture experiments, the synbiotic combination of U. rigida and L. reuteri KUB-AC5 demonstrated the potential to eliminate Salmonella Typhimurium DMST 48437 when inoculated at 104 CFU/mL within 9 h. The synbiotic activities of U. rigida and L. reuteri KUB-AC5 were further investigated using an in vitro human gut model. Compared to the probiotic treatment, the synbiotic combination of L. reuteri KUB-AC5 and U. rigida showed significantly higher levels of L. reuteri KUB-AC5 (5.1 log copies/mL) and a reduction of S. Typhimurium by 0.8 log (CFU/ml) after 24 h (p ≤ 0.05). Synbiotic treatment also significantly promoted the production of short-chain fatty acids (SCFAs), including butyric acid, propionic acid, and acetic acid, compared to prebiotic and probiotic treatments alone (p ≤ 0.05). Furthermore, the synbiotic formulation modulated the in vitro simulated gut microbiome, enhancing putatively beneficial gut microbes, including lactobacilli, Faecalibacterium, and Blautia. Our findings demonstrated that L. reuteri KUB-AC5, in combination with U. rigida, exhibited synergistic activity, as indicated by increased viability, higher anti-pathogenicity toward Salmonella, and the ability to modulate the gut microbiome.

Microencapsulating role of whey protein isolate and sucrose in protecting the cell membrane and enhancing survival of probiotic lactobacilli strains during spray drying, storage, and simulated gastrointestinal passage.

Limosilactobacillus reuteriKUB-AC5 andLactobacillus johnsoniiKUNN19-2 were spray-dried using whey protein isolate, sucrose, and whey protein isolate combined with sucrose at 130 °C and 150 °C. Differences in membrane fatty acid compositions of probiotics led to differences in cell membrane fluidity, cell injury, and cell survival during spray drying. Combination of whey protein isolate and sucrose improved survival of L. reuteriKUB-AC5, whose cell membrane were more fluid and more sensitive to stress, while whey protein isolate provided the best protection for L. johnsoniiKUNN19-2 against changes in protein secondary structures. Moreover, a relationship was found between membrane damage and the shelf-life ofmicrocapsules. Both microencapsulated cells prepared at the inlet temperature of 130 °C showed an enhanced survival upon simulated gastric digestion compared to the free cells. A thorough understanding of injury sites during spray drying will contribute to establishing optimal conditions and encapsulant formulations that maintain the cells' viability during spray drying, storage, and gastrointestinal passage.

Selection of pretreatment method and mannanase enzyme to improve the functionality of palm kernel cake.

Palm kernel cake (PKC) is a by-product of palm kernel oil extraction with moderate nutritional value, containing 30-35% ß-mannan, which is indigestible, slows growth, and reduces feed efficiency. PKC can be improved by mannanase hydrolysis, but the effectiveness of mannanase is dependent on the microbial source. Thus, the effect of steam pretreatment and bacterial mannanases on PKC quality was investigated. PKC was pretreated by steaming and hydrolyzed in the small intestine by various mannanases. The contents of reducing sugar, total sugar, and protein release were measured. Steamed PKC had a significant increase in protein (16.95 ± 0.14 to 20.98 ± 0.13%) and a substantial decrease in hemicellulose (29.52 ± 0.44 to 3.46 ± 0.88%) and lignin (8.94 ± 0.28 to 1.40 ± 0.22%). Mannanases from Escherichia coli-KMAN-3 and E. coli-Man6.7 recorded the highest activities, followed by commercial mannanase, Bacillus circulans NT6.7 and B. amyloliquefaciens NT6.3 mannanases, orderly. B. circulans NT6.7 and B. amyloliquefaciens NT6.3 had multi-activities that include glucanase (3.10 ± 0.04% and 2.47 ± 0.02%) and amylase (1.74 ± 0.03% and 1.38 ± 0.04%), respectively. B. amyloliquefaciens NT6.3 mannanase hydrolyzed steamed PKC to release more reducing sugar, total sugar, and protein than hydrolyzed raw PKC. In raw and steamed PKC, B. amyloliquefaciens NT6.3 mannanase produced the highest reducing sugar release. As a result, steam pretreatment and mannanase hydrolysis, particularly from B. amyloliquefaciens, can be used to increase the functioning of PKC and develop new feed ingredients for monogastric animals at a reasonable cost.

Probing Genome-Scale Model Reveals Metabolic Capability and Essential Nutrients for Growth of Probiotic Limosilactobacillus reuteri KUB-AC5.

Limosilactobacillus reuteri KUB-AC5 displays the hallmark features of probiotic properties for food and feed industries. Optimization of cultivation condition for the industrial production is important to reach cell concentration and cost reduction. Considering the strain-specific growth physiology, metabolic capability, and essential nutrients of L. reuteri KUB-AC5, the genome-scale metabolic model (GSMM) of L. reuteri KUB-AC5 was developed. Hereby, the GSMM of iTN656 was successfully constructed which contained 656 genes, 831 metabolites, and 953 metabolic reactions. The iTN656 model could show a metabolic capability under various carbon sources and guide potentially 14 essential single nutrients (e.g., vitamin B complex and amino acids) and 2 essential double nutrients (pairwise glutamine-glutamate and asparagine-aspartate) for L. reuteri KUB-AC5 growth through single and double omission analysis. Promisingly, the iTN656 model was further integrated with transcriptome data suggesting that putative metabolic routes as preferable paths e.g., sucrose uptake, nucleotide biosynthesis, urea cycle, and glutamine transporter for L. reuteri KUB-AC5 growth. The developed GSMM offers a powerful tool for multi-level omics analysis, enabling probiotic strain optimization for biomass overproduction on an industrial scale.

Selection of potential probiotics with cholesterol-lowering properties for probiotic yoghurt production.

From 61 lactic acid bacteria (LAB) isolates, three had good cholesterol-lowering properties, with Limosilactobacillus fermentum KUB-D18 having the highest cholesterol assimilation (68.75%) (51 µg/109 CFU). In addition, Lactiplantibacillus pentosus HM04-25 and L. pentosus HM04-3 had the two highest levels of bile salt hydrolase (BSH) activity (22.60 and 21.45 U/mL, respectively). These three strains could resist four antibiotics (aztreonam, vancomycin, teicoplanin, and nalidixic). However, fortunately, they contained no mobile antibiotic resistance genes. To evaluate the influence of probiotic strains in yoghurt production, L. fermentum KUB-D18, L. pentosus HM04-25, or L. pentosus HM04-3 were simultaneously cultured with commercial yoghurt starter (YF-L812) and incubated at 43 °C for 6 h. During yoghurt fermentation, the total bacteria in the yoghurt tended to increase from 7.39 to 8.90 log CFU/mL. The growth rates of two probiotic strains (L. pentosus HM04-25 and L. pentosus HM04-3) were stable at 6.06 to 6.62 log CFU/mL. Only the rate for L. fermentum KUB-D18 increased (to 7.5 log CFU/mL). These three probiotics did not affect the physical characteristics of yoghurt. The total soluble solids, pH, and titratable acidity values of the probiotic yoghurts were similar to the control yoghurt at 30°Brix, 4.91, and 0.90%, respectively. The firmness values of the probiotic yoghurts and the control were not significantly different (p > 0.05). Differentiation of the appearance of color, odor, flavor, and texture between the control yoghurt and the probiotic yoghurts was investigated using 56 volunteers and no significant differences were identified. Additionally, sensory evolution revealed that the acceptability of the probiotic yoghurts was higher than for the control (p ≤ 0.05). Therefore, the three probiotic strains with cholesterol-lowering properties had potential in future yoghurt production.

Crystallization, structural characterization and kinetic analysis of a GH26 ß-mannanase from Klebsiella oxytoca KUB-CW2-3.

ß-Mannanase (EC 3.2.1.78) is an enzyme that cleaves within the backbone of mannan-based polysaccharides at ß-1,4-linked D-mannose residues, resulting in the formation of mannooligosaccharides (MOS), which are potential prebiotics. The GH26 ß-mannanase KMAN from Klebsiella oxytoca KUB-CW2-3 shares 49-72% amino-acid sequence similarity with ß-mannanases from other sources. The crystal structure of KMAN at a resolution of 2.57 Šrevealed an open cleft-shaped active site. The enzyme structure is based on a (ß/α)8-barrel architecture, which is a typical characteristic of clan A glycoside hydrolase enzymes. The putative catalytic residues Glu183 and Glu282 are located on the loop connected to ß-strand 4 and at the end of ß-strand 7, respectively. KMAN digests linear MOS with a degree of polymerization (DP) of between 4 and 6, with high catalytic efficiency (kcat/Km) towards DP6 (2571.26 min-1 mM-1). The predominant end products from the hydrolysis of locust bean gum, konjac glucomannan and linear MOS are mannobiose and mannotriose. It was observed that KMAN requires at least four binding sites for the binding of substrate molecules and hydrolysis. Molecular docking of mannotriose and galactosyl-mannotetraose to KMAN confirmed its mode of action, which prefers linear substrates to branched substrates.

A randomized trial to evaluate the impact of copra meal hydrolysate on gastrointestinal symptoms and gut microbiome.

The impact of copra meal hydrolysate (CMH) on gut health was assessed by conducting a double-blinded, placebo-controlled study. Sixty healthy adult participants, aged 18-40 years were assigned to daily consume 3 g of CMH, 5 g of CMH or placebo in the form of drink powder for 21 days. Consumption of CMH at 3 g/d improved defecating conditions by reducing stool size and also relieved flatulence and bloating symptoms. Fecal samples were collected serially at the baseline before treatment, after the treatment and after a 2-week washout period. The gut microbiomes were similar among the treatment groups, with microbial community changes observed within the groups. Intake of CMH at 3 g/d led to increase microbial diversity and richness. Reduction of the ratio between Firmicutes to Bacteroidetes was observed, although it was not significantly different between the groups. The 3 g/d CMH treatment increased beneficial microbes in the group of fiber-degrading bacteria, especially human colonic Bacteroidetes, while induction of Bifidobacteriaceae was observed after the washout period. Intake of CMH led to increase lactic acid production, while 3 g/d supplement promoted the present of immunoglobulin A (IgA) in stool samples. The 3 g daily dose of CMH led to the potentially beneficial effects on gut health for healthy individuals.

Integrative growth physiology and transcriptome profiling of probiotic Limosilactobacillus reuteri KUB-AC5.

Limosilactobacillus reuteri KUB-AC5 has been widely used as probiotic in chicken for Salmonella reduction. However, a preferable carbon source and growth phase is poorly characterized underlying metabolic responses on growth and inhibition effects of L. reuteri KUB-AC5. This study therefore aimed to investigate transcriptome profiling of L. reuteri KUB-AC5 revealing global metabolic responses when alteration of carbon sources and growth phases. Interestingly, L. reuteri KUB-AC5 grown under sucrose culture showed to be the best for fast growth and inhibition effects against Salmonella Enteritidis S003 growth. Towards the transcriptome profiling and reporter proteins/metabolites analysis, the results showed that amino acid transport via ABC systems as well as sucrose metabolism and transport are key metabolic responses at Logarithmic (L)-phase of L. reuteri KUB-AC5 growth. Considering the Stationary (S)-phase, we found the potential reporter proteins/metabolites involved in carbohydrate metabolism e.g., levansucrase and levan. Promisingly, levansucrase and levan were revealed to be candidates in relation to inhibition effects of L. reuteri KUB-AC5. Throughout this study, L. reuteri KUB-AC5 had a metabolic control in acclimatization to sucrose and energy pools through transcriptional co-regulation, which supported the cell growth and inhibition potentials. This study offers a perspective in optimizing fermentation condition through either genetic or physiological approaches for enhancing probiotic L. reuteri KUB-AC5 properties.

Anti-inflammatory Effect of Probiotic Limosilactobacillus reuteri KUB-AC5 Against Salmonella Infection in a Mouse Colitis Model.

Acute non-typhoidal salmonellosis (NTS) caused by Salmonella enterica Typhimurium (STM) is among the most prevalent of foodborne diseases. A global rising of antibiotic resistance strains of STM raises an urgent need for alternative methods to control this important pathogen. Major human food animals which harbor STM in their gut are cattle, swine, and poultry. Previous studies showed that the probiotic Limosilactobacillus (Lactobacillus) reuteri KUB-AC5 (AC5) exhibited anti-Salmonella activities in chicken by modulating gut microbiota and the immune response. However, the immunobiotic effect of AC5 in a mammalian host is still not known. Here, we investigated the anti-Salmonella and anti-inflammatory effects of AC5 on STM infection using a mouse colitis model. Three groups of C57BL/6 mice (prophylactic, therapeutic, and combined) were fed with 109 colony-forming units (cfu) AC5 daily for 7, 4, and 11 days, respectively. Then, the mice were challenged with STM compared to the untreated group. By using a specific primer pair, we found that AC5 can transiently colonize mouse gut (colon, cecum, and ileum). Interestingly, AC5 reduced STM gut proliferation and invasion together with attenuated gut inflammation and systemic dissemination in mice. The decreased STM numbers in mouse gut lumen, gut tissues, and spleen possibly came from longer AC5 feeding duration and/or the combinatorial (direct and indirect inhibitory) effect of AC5 on STM. However, AC5 attenuated inflammation (both in the gut and in the spleen) with no difference between these three approaches. This study demonstrated that AC5 confers both direct and indirect inhibitory effects on STM in the inflamed gut.

Anti-cancer and anti-inflammatory effects elicited by short chain fatty acids produced by Escherichia coli isolated from healthy human gut microbiota.

BACKGROUND: Extracellular metabolites of short chain fatty acids (SCFA) excreted by gut microbiota have been reported to play an important role in the regulation of intestinal homeostasis. Apart from supplying energy, SCFA also elicit immune stimulation in animal and human cells. Therefore, an attempt was conducted to isolate SCFA producing bacteria from healthy human microbiota. The anti-cancer and anti-inflammatory effects of extracellular metabolites and individual SFCA were further investigated by using breast, colon cancer and macrophage cells. Toxin, inflammatory and anti-inflammatory cytokine gene expressions were investigated by RT-qPCR analyses in this study. RESULTS: Escherichia coli KUB-36 was selected in this study since it has the capability to produce seven SCFA extracellularly. It produced acetic acid as the main SCFA. It is a non-exotoxin producer and hence, it is a safe gut microbiota. The IC50 values indicated that the E. coli KUB-36 metabolites treatment elicited more potent cytotoxicity effect on MCF7 breast cancer cell as compared to colon cancer and leukemia cancer cells but exhibited little cytotoxic effects on normal breast cell. Furthermore, E. coli KUB-36 metabolites and individual SCFA could affect inflammatory responses in lipopolysaccharide-induced THP-1 macrophage cells since they suppressed inflammatory cytokines IL-1ß, IL-6, IL-8 and TNF-α well as compared to the control, whilst inducing anti-inflammatory cytokine IL-10 expression. CONCLUSION: SCFA producing E. coli KUB-36 possessed vast potential as a beneficial gut microbe since it is a non-exotoxin producer that exhibited beneficial cytotoxic effects on cancer cells and elicited anti-inflammatory activity simultaneously. However, the probiotic characteristic of E. coli KUB-36 should be further elucidated using in vivo animal models.

Analysis of the infant gut microbiome reveals metabolic functional roles associated with healthy infants and infants with atopic dermatitis using metaproteomics.

The infant gut microbiome consists of a complex and diverse microbial community. Comprehensive taxonomic and metabolic functional knowledge about microbial communities supports medical and biological applications, such as fecal diagnostics. Among the omics approaches available for the investigation of microbial communities, metaproteomics-based analysis is a very powerful approach; under this method, the activity of microbial communities is explored by investigating protein expression within a sample. Through use of metaproteomics, this study aimed to investigate the microbial community composition of the infant gut to identify different key proteins playing metabolic functional roles in the microbiome of healthy infants and infants with atopic dermatitis in a Thai population-based birth cohort. Here, 18 fecal samples were analyzed by liquid chromatography-tandem mass spectrometry to conduct taxonomic, functional, and pathway-based protein annotation. Accordingly, 49,973 annotated proteins out of 68,232 total proteins were investigated in gut microbiome samples and compared between the healthy and atopic dermatitis groups. Through differentially expressed proteins (DEPs) analysis, 130 significant DEPs were identified between the healthy and atopic dermatitis groups. Among these DEPs, eight significant proteins were uniquely expressed in the atopic dermatitis group. For instance, triosephosphate isomerase (TPI) in Bifidobacteriaceae in the genus Alloscardovia and demethylmenaquinone methyltransferase (DMM) in Bacteroides were shown to potentially play metabolic functional roles related to disease. PPI network analysis revealed seven reporter proteins showing metabolic alterations between the healthy and disease groups associated with the biosynthesis of ubiquinone and other quinones as well as the energy supply. This study serves as a scaffold for microbial community-wide metabolic functional studies of the infant gut microbiome in relation to allergic disease.

Lactococcus lactis KA-FF 1-4 reduces vancomycin-resistant enterococci and impacts the human gut microbiome.

Probiotic is an alternative method to treat intestinal infection disease caused by antibiotic-resistant bacteria. In this study, Lactococcus lactis KA-FF 1-4 demonstrated to have the potential to inhibit the growth of Vancomycin-resistant enterococci (VRE) by producing anti-microbial substance. In co-culture, L. lactis KA-FF 1-4 (108 CFU/mL) inhibited the growth of VRE from 103-104 CFU/mL to zero after 6 h of exposure. However, in a gut model contained human gut microbiota, this anti-VRE activity of L. lactis KA-FF 1-4 was reduced to only 3.59-6.12%. The unexpected difference in efficacy between the experimental models could be explained by the fact that the growth of L. lactis KA-FF 1-4 was stable in the gut model. Leaving aside these limitations, we observed that adding L. lactis KA-FF 1-4 into the human gut model containing VRE was able to enhance microbial richness and diversity. Specifically, a higher abundance of beneficial microbes from the group of Bifidobacterium spp. and Bacteroides fragilis. L. lactis KA-FF 1-4 also enhanced the abundance of Parabacteroides, Lactococcus, and Fusobacterium and promoted the production of lactic acid in the gut model. However, these effects were not observed in the gut model without L. lactis KA-FF 1-4. Even though this study could not demonstrate a significant anti-VRE effect of the L. lactis KA-FF 1-4 in a gut model, our results still offer evidence that L. lactis KA-FF 1-4 could positively modulate the gut microbiota by promoting the growth of beneficial microbes and their metabolite. L. lactis KA-FF 1-4 has probiotic properties to fight against VRE infection, therefore further investigation in animal model is needed.

Promising discovery of beneficial Escherichia coli in the human gut.

Ingested dietary fibres are hydrolysed by colon microbiota to produce energy-providing short-chain fatty acids (SCFA) that stimulate anti-inflammatory effects. SCFA-producing bacteria were screened from bacteria isolated from human faeces using bromothymol blue as an acid indicator and gas chromatography for SCFA profiling. The beneficial functions (antagonistic activity, haemolytic activities, antibiotic susceptibility, mucus adherent percentage and toxin gene detection) were evaluated for the top five SCFA-producing bacteria isolated from three healthy volunteers that identified as Escherichia coli strains. They produced acetic, propionic, isobutyric, butyric, isovaleric, valeric and caproic acids at average concentrations of 15.9, 1.8, 1.1, 1.9, 1.8, 2.7 and 3.4 mM, respectively. The SCFA production by E. coli strains was rapidly increased during the first 8 h of incubation and gradually decreased after 16 h of incubation. All E. coli strains showed acid and bile tolerance, resulting in a survival rate greater than 70% with no haemolytic activity, mucus adherence greater than 40% and susceptibility to conventional antibiotics. Hence, the selected E. coli strains exhibited promising probiotic properties with neither enterotoxin nor LPS producibility was detected. The present results confirm the existence of friendly and harmless E. coli strains in human microbiota as potential probiotics.

Age-related changes in the gut microbiota and the core gut microbiome of healthy Thai humans.

The gut microbial diversity of Thai people was investigated between two large cohorts, adult and elderly subjects, from the middle region of Thailand; the cohorts were divided into different age groups of healthy adult (73) and elderly subjects (47). The diversities of the groups were characterized using a pyrosequencing technique with primers targeting the V6-V8 region of the 16S rRNA gene, and a significant decrease in the Firmicutes and Bacteroidetes ratio from 7.3 to 4.5 was observed with increased age. The microbiota of the adult and elderly groups had a significantly higher abundance of the phylum Actinobacteria, including the three species Bifidobacterium adolescentis, Bifidobacterium longum and Bifidobacterium pseudocatenulatum, and the phylum Bacteroidetes containing the four species Bacteroides uniformis, Bacteroides ovatus, Bacteroides caccae and Bacteroides thetaiotaomicron. Firmicutes showed no significant differences between the two groups. Eleven species belonging to Firmicutes, Bacteroidetes and Proteobacteria were shared by at least 90% of all subjects and defined as core gut microbiota of healthy Thai, among which a high abundance of Escherichia coli was particularly characterized in Thai elderly individuals. Multiple linear regression analysis of age, gender, BMI and diet consumption frequency showed the correlation of age with Bacteroides and Bifidobacterium. Rice consumption frequency showed a significant positive correlation with Bacteroides, while no correlation was found for other factors. Taken together, in the gut of Thai adults, Bifidobacterium decreased and Bacteroides increased with age, while rice consumption increased the abundance of Bacteroides. These link of age and food, especially rice carbohydrate, to gut microbiota and health could be ultimately proposed as the Thai feature.

Copra meal hydrolysis by the recombinant ß-mannanase KMAN-3 and MAN 6.7 expressed in Escherichia coli.

Hydrolysis products of defatted copra meal (DCM) hydrolysis were investigated with either recombinant ß-mannanases from Klebsiella oxytoca KUB-CW2-3 (KMAN-3) or Bacillus circulans NT 6.7 (MAN 6.7). Morphological changes and functional groups of solid residues were also determined by scanning electron microscopy (SEM) and Fourier transform infrared (FTIR) spectroscopy. Results revealed that the Michaelis-Menten constant (K m) and maximum velocity (V max) values of KMAN-3 on DCM were 2.4 mg/ml and 5.4 U/mg, respectively, while MAN 6.7 recorded K m and V max at 2.0 mg/ml and 4.3 U/mg, respectively. Both enzymes efficiently randomly hydrolysed DCM and produced a range of different manno-oligosaccharides (MOS). The profile of hydrolysis products was different for each enzyme used. Main products from hydrolysis of DCM by KMAN-3 and MAN 6.7 were various MOS including mannobiose (M2), mannotriose (M3), mannotetraose (M4), and mannose, whereas mannopentaose (M5) was only found from KMAN-3. Amount of M3 produced by KMAN-3 was about three times higher than from MAN 6.7. Total MOS yield for KMAN-3 was 1.5-folds higher than for MAN 6.7. SEM analysis showed that enzymatic hydrolysis with KMAN-3 and MAN 6.7 resulted in deconstruction of the DCM structure which generated a variety of MOS products. FTIR spectra revealed that the properties of both hydrolysed solids were not significantly different compared to the original DCM. Results suggested that KMAN-3 was a promising candidate for production of high MOS content from copra meal.

Characterization and probiotic properties of Lactobacilli from human breast milk.

Several studies have reported a complex microbial community in human breast milk. This community impacts the shape of the infant gut microbiota and consequently impacts host health. Lactobacillus is an important probiotic and has many applications in the functional food industry. This study isolated and evaluated the potential probiotic bacteria from human milk. Two Lactobacillus species, L. plantarum and L. pentosus, were isolated from the breast milk of Thai women. L. pentosus HM04-22, L. pentosus HM04-3, L. plantarum HM04-80, L. plantarum HM04-88 and L. plantarum HM01-1 showed good adhesion activity (> 55%) and resistance in gastric (pH 2) and bile (pH 8) conditions. Characterization of the probiotic properties indicated that all selected Lactobacillus isolates had anti-adhesion properties against Escherichia coli and Salmonella Typhimurium. Lactobacillus isolates protected Caco-2 cells from pathogen adhesion at 25-40%. In addition, the five selected strains presented anti-inflammatory properties by reducing interleukin (IL)-8 expression at 0.14 ± 0.16 to 0.52 ± 0.117-fold. However, the strains had no effect on the expression of tight junction genes, including zona occludens (ZO)-1, occludin and claudin-1. In conclusion, five selected Lactobacillus isolates from human milk were candidates for use as probiotics to promote health. However, more tests in animal models and clinical trials need to be performed.

Author Correction: Diversity in gut bacterial community of school-age children in Asia.

A correction has been published and is appended to both the HTML and PDF versions of this paper. The error has not been fixed in the paper.

In vitro fermentation of copra meal hydrolysate by human fecal microbiota.

Copra meal hydrolysate (CMH) is obtained by hydrolyzing defatted copra meal with ß-mannanase from Bacillus circulans NT 6.7. In this study, we investigated the resistance of CMH to upper gastrointestinal tract digestion and the fecal fermentation profiles of CMH. Fecal slurries from four healthy human donors were used as inocula, and fructooligosaccharides (FOS) were used as a positive prebiotic control. Fecal batch cultures were performed at 37 °C under anaerobic conditions. Samples were collected at 0, 10, 24 and 34 h for bacterial enumeration via fluorescent in situ hybridization and organic acid (OA) analysis. In vitro gastric stomach and human pancreatic α-amylase simulations demonstrated that CMH was highly resistant to hydrolysis. Acetate was the main fermentation product of all the substrates. The proportions of acetate production of the total OAs from FOS, CMH and yeast mannooligosaccharides (MOS) after 34 h of fermentation did not significantly differ (69.76, 65.24 and 53.93%, respectively). At 24 h of fermentation, CMH promoted the growth of Lactobacillus and Bifidobacterium groups (P < 0.01) and did not significantly differ from the results obtained using FOS. The results of in vitro fecal fermentation of CMH indicate that CMH can promote the growth of beneficial bacteria.

Improving palm kernel cake nutrition using enzymatic hydrolysis optimized by Taguchi method.

Enzymatic hydrolysis of palm kernel cake to improve the quality of substrates with multi-response criteria based on the Taguchi orthogonal array. Nine experimental runs were performed based on an L9 orthogonal array. Percent substrate, incubation time, and enzyme units were optimized considering multiple performance characteristics. Analysis of variance was also applied to identify the most significant factors. Results determined percent substrate as the most important factor for enzymatic hydrolysis followed by incubation time and enzyme units. Enzymatic hydrolysis conditions were optimized as percent substrate, incubation time and enzyme units at 14%, 6 h and 750 units, respectively. Tests were conducted to compare experimental and model results. The experimental result (protein release) at optimal condition were three times higher than the predicted mode.