Distinct gut microbiota of healthy children from two different geographic regions of Thailand.

In Thailand, food consumption by people from each region is different. This can be an important environmental factor which shapes the gut microbiota further affecting their health. This study aimed to use quantitative PCR (qPCR) to investigate the intestinal microbial community in 60 healthy children (aged 8-11 years) living in specific areas, namely central (CT) and northeastern (NE) Thailand where each region has its own typical food consumption. The children from NE had significantly higher consumption frequency of meat (chicken and beef), a wide variety of carbohydrate sources (noodle, fermented rice and sweet potato) including vegetables and fruit, while in CT, there was a significant preference for rice, breakfast cereal and cow milk. The qPCR analysis resulted in significantly higher abundance of lactobacilli, Clostridium coccoides-Eubacterium rectale, Clostridium leptum, Prevotella and Bacteroides fragilis in children from the NE region. However, no significant difference in the count of Bifidobacterium spp., Enterobacteriaceae and methanogens was observed. Considering the correlation of food sources and microbial groups, the consumption frequency of vegetables showed a moderately positive correlation coefficient of 0.42 and 0.34 to the Lactobacillus group (P = 0.001) and the Prevotella group (P = 0.008), respectively, while a diet of fish and beef showed a moderately negative correlation coefficient of -0.41 (P = 0.001) and -0.33 (P = 0.09) to Bifidobacterium spp., respectively. Our results suggested that high frequency consumption of varieties of carbohydrates, protein sources, fruits and vegetables by the NE children promoted a high abundance of bacterial species in the phyla Firmicutes and Bacteroidetes.

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.

Evaluating the Prebiotic Properties of Agar Oligosaccharides Obtained from the Red Alga Gracilaria fisheri via Enzymatic Hydrolysis.

Currently, the demand in the food market for oligosaccharides with biological activities is rapidly increasing. In this study, agar polysaccharides from Gracilaria fisheri were treated with ß-agarases and hydrolyzed to agar oligosaccharides (AOSs). High-performance anion-exchange chromatography/pulsed amperometric detection (HPAEC-PAD), Fourier-transform infrared spectroscopy (FT-IR), and gel permeation chromatography (GPC), were employed to analyze the chemical characteristics of AOSs. The FT-IR spectra revealed that the enzymatic hydrolysis had no effect on specific functional groups in the AOS molecule. To investigate the prebiotic and pathogen inhibitory effects of AOSs, the influence of AOSs on the growth of three probiotic and two pathogenic bacteria was examined. The gastrointestinal tolerance of probiotics in the presence of AOSs was also investigated. AOSs enhanced the growth of Lactobacillus plantarum by 254%, and inhibited the growth of Bacillus cereus by 32.80%, and Escherichia coli by 58.94%. The highest survival rates of L. plantarum and L. acidophilus were maintained by AOSs in the presence of α-amylase and HCl under simulated gastrointestinal conditions. This study demonstrates that AOSs from G. fisheri exhibit potential as a prebiotic additive in foods.

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.

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.

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

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Urban Diets Linked to Gut Microbiome and Metabolome Alterations in Children: A Comparative Cross-Sectional Study in Thailand.

Loss of traditional diets by food globalization may have adverse impact on the health of human being through the alteration of gut microbial ecosystem. To address this notion, we compared the gut microbiota of urban (n = 17) and rural (n = 28) school-aged children in Thailand in association with their dietary habits. Dietary records indicated that children living in urban Bangkok tended to consume modern high-fat diets, whereas children in rural Buriram tended to consume traditional vegetable-based diets. Sequencing of 16S rRNA genes amplified from stool samples showed that children in Bangkok have less Clostridiales and more Bacteroidales and Selenomonadales compared to children in Buriram and bacterial diversity is significantly less in Bangkok children than in Buriram children. In addition, fecal butyrate and propionate levels decreased in Bangkok children in association with changes in their gut microbial communities. Stool samples of these Thai children were classified into five metabolotypes (MTs) based on their metabolome profiles, each characterized by high concentrations of short and middle chain fatty acids (MT1, n = 17), amino acids (MT2, n = 7), arginine (MT3, n = 6), amino acids, and amines (MT5, n = 8), or an overall low level of metabolites (MT4, n = 4). MT1 and MT4 mainly consisted of samples from Buriram, and MT2 and MT3 mainly consisted of samples from Bangkok, whereas MT5 contained three samples from Bangkok and five from Buriram samples. According to the profiles of microbiota and diets, MT1 and MT2 are characteristic of children in Buriram and Bangkok, respectively. Predicted metagenomics indicated the underrepresentation in MT2 of eight genes involved in pathways of butyrate biosynthesis, notably including paths from glutamate as well as pyruvate. Taken together, this study shows the benefit of high-vegetable Thai traditional diets on gut microbiota and suggests that high-fat and less-vegetable urban dietary habits alter gut microbial communities in Thai children, which resulted in the reduction of colonic short chain fatty acid fermentation.

Microbial Community of Healthy Thai Vegetarians and Non-Vegetarians, Their Core Gut Microbiota, and Pathogen Risk.

Pyrosequencing analysis of intestinal microflora from healthy Thai vegetarians and non-vegetarians exhibited 893 OTUs covering 189 species. The strong species indicators of vegetarians and non-vegetarians were Prevotella copri and Bacteroides vulgatus as well as bacteria close to Escherichia hermanii with % relative abundance of 16.9 and 4.5-4.7, respectively. Core gut microbiota of the vegetarian and non-vegetarian groups consisted of 11 and 20 different bacterial species, respectively, belonging to Actinobacteria, Firmicutes, and Proteobacteria commonly found in both groups. Two species, Faecalibacterium prausnitzii and Gemmiger formicilis, had a prevalence of 100% in both groups. Three species, Clostridium nexile, Eubacterium eligens, and P. copri, showed up in most vegetarians, whereas more diversity of Collinsella aerofaciens, Ruminococcus torques, various species of Bacteroides, Parabacteroides, Escherichia, and different species of Clostridium and Eubacterium were found in most non-vegetarians. Considering the correlation of personal characters, consumption behavior, and microbial groups, the age of non-vegetarians showed a strong positive correlation coefficient of 0.54 (p = 0.001) to Bacteroides uniformis but exhibited a moderate one to Alistipes finegoldii and B. vulgatus. Only a positive moderate correlation of body mass index and Parabacteroides distasonis appeared. Based on the significant abundance of potential pathogens, the microbiota of the non-vegetarian group showed an abundance of potential pathogen varieties of Bilophila wadsworthia, Escherichia coli, and E. hermannii, whereas that of the vegetarian group served for only Klebsiella pneumoniae. These results implied that the microbiota of vegetarians with high abundance of P. copri and low potential pathogen variety would be a way to maintain good health in Thais.

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

Asia differs substantially among and within its regions populated by diverse ethnic groups, which maintain their own respective cultures and dietary habits. To address the diversity in their gut microbiota, we characterized the bacterial community in fecal samples obtained from 303 school-age children living in urban or rural regions in five countries spanning temperate and tropical areas of Asia. The microbiota profiled for the 303 subjects were classified into two enterotype-like clusters, each driven by Prevotella (P-type) or Bifidobacterium/Bacteroides (BB-type), respectively. Majority in China, Japan and Taiwan harbored BB-type, whereas those from Indonesia and Khon Kaen in Thailand mainly harbored P-type. The P-type microbiota was characterized by a more conserved bacterial community sharing a greater number of type-specific phylotypes. Predictive metagenomics suggests higher and lower activity of carbohydrate digestion and bile acid biosynthesis, respectively, in P-type subjects, reflecting their high intake of diets rich in resistant starch. Random-forest analysis classified their fecal species community as mirroring location of resident country, suggesting eco-geographical factors shaping gut microbiota. In particular, children living in Japan harbored a less diversified microbiota with high abundance of Bifidobacterium and less number of potentially pathogenic bacteria, which may reflect their living environment and unique diet.