Effect of Tempeh and Daidzein on Calcium Status, Calcium Transporters, and Bone Metabolism Biomarkers in Ovariectomized Rats.

Menopause marks a critical life stage characterized by hormonal changes that significantly impact bone health, leading to a heightened susceptibility to bone fractures. This research seeks to elucidate the impact of daidzein and tempeh on calcium status, calcium transporters, and bone metabolism in an ovariectomized rat model. Forty female Wistar rats, aged 3 months, participated in a two-phase experiment. The initial phase involved inducing a calcium deficit, while the second phase comprised dietary interventions across five groups: Sham (S) and Ovariectomy (O) with a standard diet, O with bisphosphonate (OB), O with pure daidzein (OD), and O with tempeh (OT). Multiple parameters, encompassing calcium levels, calcium transporters, bone histopathology, and serum bone metabolism markers, were evaluated. The findings revealed that the OT group showcased heightened levels of bone turnover markers, such as pyridinoline, C-telopeptide of type I collagen, bone alkaline phosphatase, and procollagen type I N-terminal propeptide, in contrast to S and O groups, with statistical significance (p < 0.05). Histopathologically, both the OD and OT groups exhibited effects akin to the OB group, indicating a decrease in the surface area occupied by adipocytes in the femoral bone structure, although statistically non-equivalent, supporting the directionally similar trends. Although TRPV5 and TRPV6 mRNA expression levels in the jejunum and duodenum did not display statistically significant differences (p > 0.05), the OD and OT groups exhibited increased expression compared to the O group. We hypothesized that obtained results may be related to the effect of isoflavones on estrogen pathways because of their structurally similar to endogenous estrogen and weak estrogenic properties. In conclusion, the daily consumption of pure daidzein and tempeh could potentially improve and reinstate calcium status, calcium transport, and bone metabolism in ovariectomized rats. Additionally, isoflavone products demonstrate effects similar to bisphosphonate drugs on these parameters in ovariectomized rats.

Influence of supplementation with iron and probiotic bacteria Lactobacillus plantarum and Lactobacillus curvatus on selected parameters of inflammatory state in rats on a high-fat iron-deficient diet.

BACKGROUND: A high-fat (HF) diet, diet iron deficiency and iron supplementation may affect inflammatory parameters. Probiotics influence both iron metabolism and inflammation. We compared the inflammatory state in rats on a HF iron-deficient diet receiving oral iron, Lactobacillus plantarum and Lactobacillus curvatus in different combinations. METHODS: This was a two-stage experiment. In groups C (n = 8) and HF (n = 8), rats ate a control or HF diet, respectively, for 16 weeks. In the group HFDEF (n = 48), rats ate a HF iron-deficient diet for 8 weeks (first stage) and were subsequently divided into 6 groups (n = 8 each) receiving the following for a further 8 weeks (second stage): HFDEF - a HF iron-deficient diet; HFDEFFe - a HF iron-deficient diet with iron; HFDEFLp and HFDEFLc - a HF iron-deficient diet with L. plantarum or L. curvatus, respectively; and HFDEFFeLp and HFDEFFeLc - a HF iron-deficient diet with iron and L. plantarum or L. curvatus, respectively. Body composition analysis and blood sampling was performed. Markers of iron status and levels of total antioxidant status (TAS), C-reactive protein (CRP), tumour necrosis factor alpha (TNF-α) and interleukin 6 (IL-6) were measured in the blood. RESULTS: TAS was higher in the HFDEF group (756.57 ± 489.53 ng mL-1) versus the HFDEFLc group (187.04 ± 47.84 ng mL-1; P = 0.022). No more differences were found between groups, or in TAS, CRP, TNF-α and IL-6 concentrations. Also, no differences were found between groups for alanine and aspartate aminotransferases, glucose, total cholesterol, low- and high-density lipoproteins and triglycerides. TAS level was positively correlated with ferritin concentration, IL-6 with TAS and TNF-α with hepcidin level. CONCLUSIONS: Supplementation with L. plantarum, L. curvatus and iron in combinations exerts no influence on inflammatory status, lipid profile, hepatic function and serum fasting glucose in rats on a HF iron-deficient diet. © 2024 Society of Chemical Industry.

Effects of Daidzein, Tempeh, and a Probiotic Digested in an Artificial Gastrointestinal Tract on Calcium Deposition in Human Osteoblast-like Saos-2 Cells.

Adequate calcium intake is crucial for the prevention and treatment of bone-related issues. Developing a nutritional source of readily bioavailable calcium is particularly significant for individuals deficient in this essential element and at risk of developing osteoporosis. This research aimed to evaluate the impact of tempeh (T), daidzein (D), and Lactobacillus acidophilus (LA) within a simulated intestinal environment consisting of Caco-2 epithelial and Saos-2 cells, focusing on their implications for bone mineralization mechanisms. In the initial phase, calcium bioaccessibility from calcium citrate (CaCt), LA, D, the daidzein combination D-CaCt-LA (D1:1:1), and the tempeh combination T-CaCt-LA (T1:1:1) was assessed through digestion simulation. The calcium content of both untreated and digested samples was determined using atomic absorption spectrometry (AAS). In the subsequent stage, the digested samples were used to induce intestinal absorption in differentiated enterocyte-like Caco-2 cells. The permeable fractions were then evaluated in a culture of osteoblast-like Saos-2 cells. Preliminary cellular experiments employed the MTT assay to assess cytotoxicity. The results indicated that the analyzed products did not influence the deposition of extracellular calcium in Saos-2 cells cultured without mineralization stimulators. The combined formulations of permeable fractions of digested CaCt, LA, D, and T demonstrated the capacity to enhance the proliferation of Saos-2 cells. In Saos-2 cells, D, D1:1:1, and LA showed no discernible impact on intracellular calcium accumulation, whereas T and T1:1:1 reduced the calcium deposits. Additionally, mRNA transcripts and alkaline phosphatase (ALP) activity levels in Saos-2 cells cultured without mineralization induction were unaffected by the analyzed products. An examination of the products revealed no discernible effect on ALP activity or mRNA expression during Saos-2 cell differentiation. Our findings suggest that tempeh, daidzein, and L. acidophilus did not positively impact cellular calcium deposition in Saos-2 cells. However, tempeh, daidzein and its combination, and L. acidophilus might enhance the process of osteogenic differentiation in Saos-2 cells. Nevertheless, this study did not identify any synergistic impact on calcium deposition and the process of osteogenic differentiation in Saos-2 cells of isoflavones and probiotics.

Influence of Isoflavones and Probiotics on Magnesium Status in Healthy Female Rats.

Isoflavones and probiotics are promising nutrients for bone health, and magnesium (Mg) is essential for bone metabolism. This study aimed to determine the effects of daidzein, genistein and Lactobacillus acidophilus on the Mg status of healthy female rats. Forty-eight rats were randomly assigned to six groups, with the control group receiving a standard diet (AIN 93M). The remaining groups were fed the same diet with added ingredients such as tempeh flour; soy flour; pure daidzein and genistein; L. acidophilus or a combination of daidzein, genistein, and L. acidophilus. Tissue samples were collected after the eight-week intervention, and Mg concentrations were analysed using flame atomic absorption spectrometry. Myeloid and erythroid cells were determined using the haematoxylin and eosin bone staining method. Statistical analyses were conducted using one-way ANOVA with Tukey's post hoc test and Pearson's correlation coefficient. The threshold for significance was p < 0.05. Compared with the control group, adding tempeh to the diet of rats resulted in significant changes in Mg concentrations in various tissues, with a decrease in the kidneys and an increase in the fur. Although not statistically significant compared to the control group, the tempeh group showed increased Mg concentrations in the femur and spleen. The myeloid-to-erythroid cell ratio did not differ significantly among groups, but all intervention groups showed higher ratios than the control group. A strong negative correlation was observed between Mg concentrations in the kidneys and fur. Conversely, a positive correlation was identified between Mg concentrations in the pancreas and fur. Daily consumption of tempeh may improve Mg status in the organism. Intake of pure daidzein, genistein, or probiotic seems to have no effect on Mg concentrations in healthy rats.

Factors Affecting the Extraction of (Poly)Phenols from Natural Resources Using Deep Eutectic Solvents Combined with Ultrasound-Assisted Extraction.

Replacing conventional solvents with deep eutectic solvents (DES) has shown promising effects on the extraction yield of (poly)phenols. DES can be combined with ultrasound-assisted extraction (UAE) to further increase the extraction efficiency of (poly)phenols from natural resources compared to conventional methods. This review discusses the factors associated with DES (composition, solvent-to-sample ratio, extraction duration, and temperature) and UAE (ultrasound frequency, power, intensity, and duty cycle) methods that influence the extraction of (poly)phenols and informs future improvements required in the optimization of the extraction process. For the optimum (poly)phenol extraction from natural resources, the following parameters shall be considered: ultrasound frequency should be in the range of 20-50 kHz, ultrasound intensity in the range of 60-120 W/cm2, ultrasound duty cycle in the range of 40-80%, ultrasound duration for 10-30 minutes, and ultrasound temperature for 25-50 °C. Among the reported DES systems, choline chloride with glycerol or lactic acid, with a solvent-to-sample mass ratio of 10-30:1 shown to be effective. The solvent composition and solvent-to-sample mass ratio should be selected according to the target compound and the source material. However, the high viscosity of DES is among the major limitations. Optimizing these factors can help to increase the yield of extracted (poly)phenols and their applications.

Effects of isoflavone and probiotic intake on calcium transport and bone metabolism biomarkers in female rats.

Calcium is essential for maintaining bone health as it contributes to bone formation, remodeling, strength, and density. This study investigated the effect of isoflavones and probiotics on calcium transporters' gene expression, serum calcium levels, and bone metabolism biomarkers in healthy female rats. Forty-eight female Wistar rats were classified into six groups. Bone metabolism biomarkers (pyridinoline, deoxypyridinoline, parathyroid hormone, and osteocalcin) and serum calcium levels were measured by enzyme-linked immunosorbent assay (ELISA) and atomic absorption spectroscopy (AAS), respectively. Gene expression of calcium transporters (Trpv5 and Trpv6) was evaluated in duodenum and jejunum tissue samples using quantitative polymerase chain reaction (qPCR). Trpv5 and Trpv6, epithelial calcium channels, play a crucial role in calcium transport and homeostasis in the body. The study consisted of a1-week adaptation period for the rats to adjust to the controlled conditions, followed by an 8-week intervention phase. The daidzein and genistein group showed a significant increase in the gene expression of the Trpv6 transporter in the duodenum and a marked decrease in serum pyridinoline levels compared to the control group. The tempeh and soybean groups showed a significant decrease in the gene expression of the Trpv5 calcium transporter in the jejunum. However, no significant influence of the Lactobacillus acidophilus diet on calcium transport and bone metabolism biomarkers was observed in the L. acidophilus group. The correlation analysis showed a significant positive relationship between serum calcium, bone metabolism biomarkers, and calcium transporters. In conclusion, our study demonstrates that the daidzein and genistein diet improves calcium transport in the duodenum and reduces pyridinoline serum concentrations, while tempeh and soybean diets reduce calcium transport in the jejunum. However, the combination of daidzein, genistein, and L. acidophilus did not demonstrate a synergistic effect on calcium transport and bone metabolism, suggesting that further investigations are needed to elucidate their potential interactions.

Corrigendum to "Isoflavones and probiotics effect on bone calcium and bone cells in rats" [Heliyon 9(6) (June 2023) e16801].

[This corrects the article DOI: 10.1016/j.heliyon.2023.e16801.].

Isoflavones and probiotics effect on bone calcium and bone cells in rats.

Isoflavones and probiotics have shown the therapeutic potential to alter calcium absorption and bone cell metabolism. This study sought to ascertain the effect of isoflavones and probiotics on calcium status and bone health in healthy female rats. Forty-eight adult female Wistar rats were grouped and fed: a standard diet (control); and standard diets with tempeh; soy; daidzein and genistein; Lactobacillus acidophilus; and a combination of daidzein, genistein, and L. acidophilus. The biochemical serum parameters, such as alanine transaminase, aspartate transaminase, glucose, and triacylglycerol concentrations, were measured, and calcium contents in tissues were determined. After staining the bone with hematoxylin and eosin, the number of osteoblasts, osteocytes, and the percentage of bone marrow adipocytes were counted. Compared with the control group, the soy group showed a significantly lower triacylglycerol concentration. The L. acidophilus group considerably increased the calcium content in the femoral bone. The daidzein and genistein, L. acidophilus, and a combination of daidzein, genistein, and L. acidophilus groups showed significantly lower calcium contents in the heart and kidneys. The daidzein and genistein group significantly enhanced the number of osteoblasts and osteocytes. A substantial inverse correlation was observed between calcium contents in kidneys and osteoblasts. In conclusion, the combination of daidzein, genistein, and L. acidophilus may improve bone calcium concentrations and bone cells. However, no synergistic effect between isoflavones and probiotics was detected in this study.

Termites as human foods-A comprehensive review.

Global food production is anticipated to rise along with the growth of the global population. As a result, creative solutions must be devised to ensure that everyone has access to nutritious, affordable, and safe food. Consequently, including insects in diets has the potential to improve global food and nutrition security. This paper aims to share recent findings by covering edible termites as the main aspect, from their consumption record until consumer acceptance. A total of 53 termite species are reported as edible ones and distributed in 6 biogeographic realms. Generally, termites have a nutrient composition that is suitable for human consumption, and cooked termites are a better dietary choice than their raw counterparts. Besides, increasing customer interest in eating termite-based food can be achieved by making it more palatable and tastier through various cooking processes, that is, boiling, frying, grilling, roasting, smoking, and sun-drying. Moreover, edible termites can also be used as a new source of medication by exhibiting antimicrobial activity. Regarding their advantages, it is strongly encouraged to implement a seminatural rearing system to sustain the supply of edible termites. Overall, this paper makes it evident that termites are an important natural resource for food or medicine. Hence, the long-term objective is to stimulate scientific inquiry into the potential of edible insects as an answer to the problem of global food security.

The impact of soy products, isoflavones, and Lactobacillus acidophilus on iron status and morphological parameters in healthy female rats.

BACKGROUND: Isoflavones and probiotics are two major factors involved in bone health. Osteoporosis and disturbances in iron (Fe) levels are common health problems in aging women. This study aimed to analyze how soybean products, daidzein, genistein, and Lactobacillus acidophilus (LA) affect Fe status and blood morphological parameters in healthy female rats. METHODS: A total of 48 Wistar rats aged 3 months were randomly divided into six groups. The control group (K) received a standard diet (AIN 93 M). The remaining five groups received a standard diet supplemented with the following: tempeh flour (TP); soy flour (RS); daidzein and genistein (DG); Lactobacillus acidophilus DSM20079 (LA); as well as a combination of daidzein, genistein, and L. acidophilus DSM20079 (DGLA). After 8 weeks of intervention, blood samples of the rats were collected for morphological analysis, whereas tissue samples were collected and kept at -80 °C until Fe analysis. Red blood cells, hemoglobin, hematocrit, mean corpuscular volume (MCV), mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration, platelets (PLTs), red cell distribution width, white blood cells, neutrophils (NEUT), lymphocytes (LYM), monocytes, eosinophils (EOS), and basophils were measured for blood morphological analysis. Fe concentrations were determined using flame atomic spectrometry. For statistical analysis, an ANOVA test for significance at the 5 % level was used. The relationship between tissue Fe levels and blood morphological parameters was determined using Pearson's correlation. RESULTS: Although no significant differences were observed in the Fe content between all diets, the TP group showed significantly higher levels of NEUT and lower levels of LYM than the control group. Compared with the DG and DGLA groups, the TP group showed a dramatically higher PLT level. In addition, the RS group showed significantly higher Fe concentrations in the spleen compared with the standard diet. Compared with the DG, LA, and DGLA groups, the RS group also showed significantly higher Fe concentrations in the liver. Compared with the TP, DG, LA, and DGLA groups, the RS group showed dramatically higher Fe concentrations in the femur. Pearson's correlations between blood morphological parameters and Fe levels in tissues were observed, especially a negative correlation between the Fe level in the femur and the NEUT concentration (-0.465) and a strong positive correlation between the Fe level in the femur and the LYM concentration (0.533). CONCLUSION: Soybean flour was found to increase Fe levels in rats, whereas tempeh may alter anti-inflammatory blood parameters. Isoflavones and probiotics did not affect Fe status in healthy female rats.

Interrelationship between Vitamin D and Calcium in Obesity and Its Comorbid Conditions.

Obesity has been linked to vitamin D (VD) deficiency and low calcium (CAL) status. In the last decade, dietary supplementation of vitamin D and calcium (VD-CAL) have been extensively studied in animal experiments and human studies. However, the physiological mechanisms remain unknown as to whether the VD-CAL axis improves homeostasis and reduces biomarkers in regulating obesity and other metabolic diseases directly or indirectly. This review sought to investigate their connections. This topic was examined in scientific databases such as Web of Science, Scopus, and PubMed from 2011 to 2021, and 87 articles were generated for interpretation. Mechanistically, VD-CAL regulates from the organs to the blood, influencing insulin, lipids, hormone, cell, and inflammatory functions in obesity and its comorbidities, such as non-alcoholic fatty liver disease, cardiovascular disease, and type-2 diabetes mellitus. Nevertheless, previous research has not consistently shown that simultaneous VD-CAL supplementation affects weight loss or reduces fat content. This discrepancy may be influenced by population age and diversity, ethnicity, and geographical location, and also by degree of obesity and applied doses. Therefore, a larger prospective cohort and randomised trials are needed to determine the exact role of VD-CAL and their interrelationship.

Fat Oxidation of Fatty Fish vs. Meat Meal Diets Under in vitro Standardized Semi-Dynamic Gastric Digestion.

Meat and fish are introduced into the diet as a source of protein, but these muscle foods present different fatty acid (FA) compositions and different lipid stabilities. Fatty fish is expected to oxidize due to its higher content of polyunsaturated FA (PUFA), whereas the higher heme-Fe content of red meat will also affect lipid stability. Combining other food ingredients within a meal also influences lipid oxidation, which will not stop after meals intake. This is due to the acidic environment of the stomach together with the presence of metallic ions, a process that is scarcely understood. The goal of this study was to evaluate the oxidation of fatty fish vs. meat meal diets under in vitro standardized semi-dynamic gastric conditions and FA release from the stomach to the duodenum. Meal diets composed by 25% beef meal (BM) or fatty fish meal (FM), 25% fried potatoes, and 50% sugar soft drink were prepared. Proximate composition, FA and amino acid profiles, and meals quality indices were evaluated. Their differences in composition led to different total gastric digestion time of 242.74 (BM) and 175.20 (FM) minutes. Using the INFOGEST semi-dynamic gastric model, 4 gastric emptying (GE) were simulated in both meals. In each GE, FA profile and lipid oxidation products (LOPs) formation were assessed. As a result, more than 50% FA release to the duodenum occurred in GE1, whose percentage decreased with the time of digestion. FM exhibited the highest LOPs formation, which corroborates the high peroxidizability index measured for this meal diet. Higher LOPs formation occurred in the later GEs, which released less FA. This suggests that higher times of residence in the stomach increase FA oxidation. This study shows a higher formation of LOPs during digestion of FM using a whole meal approach. These results relate to its richness in PUFAs compared to BM. Despite higher LOPs formation, FM digests that reached duodenum still contain higher content of unoxidized PUFAs compared with BM and a desirable ω3/ω6 PUFAs ratio of ~0.43. LOPs formation in PUFA-rich meals could be reduced if those meals have a low caloric value, avoiding large times of residence in the stomach and consequently high levels of oxidation.

Antioxidant and antibacterial activities in 21 species of Indonesian sea cucumbers.

Sea cucumbers are marine organisms with uses in food, cosmetics, and medicine. This study aimed to identify Indonesian sea cucumbers with high antioxidant and antibacterial activities. Twenty-one sea cucumber species were used for this study. Antioxidant capacity was evaluated using the 2,2-diphenyl-ß-picrylhydrazine assay. Antibacterial activity was assessed using the disk diffusion assay, whereas the resazurin-based assay was employed to determine the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). Volatile compounds possibly related to the biological activity of sea cucumbers were analyzed via gas chromatography-mass spectrometry (GC-MS). Holothuria atra had the strongest antioxidant capacity (IC50 = 14.22 ± 0.87 µg µL-1). Stichopus vastus displayed the best antibacterial activity against Staphylococcus aureus, whereas Stichopus ocellatus extract was most potent against Vibrio cholerae. Holothuria albiventer, which controlled Bacillus subtilis most effectively while also being active against S. aureus and V. cholerae, was the optimal antimicrobial species. H. albiventer and Actinopyga echinites inhibited B. subtilis growth at 12.5 µg µL-1. The MBC tests indicated that the antibacterial activities of sea cucumbers at the MIC were bacteriostatic, rather than bactericidal, in nature. GC-MS analysis uncovered long-chain fatty acids that might be associated with the antibacterial activities of sea cucumbers.

The calcium deficit diet does not affect body composition, glucose, and lipid status in ovariectomized rats.

BACKGROUND: and objectives. Ovariectomy leads to an alteration of estrogen status associated with impaired calcium metabolism, reduced bone mineral density, and disorder in lipid metabolism. Calcium deficiency may enhance the effect of estrogen-deficiency condition. Therefore, this study aimed to define whether a calcium deficit in a diet affected the body composition, glucose, and lipid blood parameters in ovariectomized rats. METHODS: This study included thirty female Wistar rats (three months old). The rats were randomly assigned into three groups: Group 1 (n = 10) consisted of rats in the control group (C) fed the standard diet, group 2 (n = 10) consisted of ovariectomized rats (OVX) fed the standard diet, and group 3 (n = 10) consisted of ovariectomized rats fed the calcium-deficit diet (OVXD). The body composition, blood glucose, and blood lipid profile were determined. RESULTS: Obtained results demonstrated that a calcium-deficient diet and a standard diet had a similar effect on feeding efficiency and body composition in ovariectomized rats. Moreover, the liver’s relative weight, heart, kidneys, pancreas, brain, and femurs significantly decreased in both ovariectomized groups. The blood levels of cholesterol (CHOL), triglycerides (TG), low-density lipoprotein cholesterol (LDL-C), and high-density lipoprotein cholesterol (HDL-C) increased similarly in the ovariectomized groups OVX and OVXD in comparison to the control group. Glucose concentration was comparable between groups. CONCLUSIONS: Calcium deficiency did not enhance the ovariectomy effect on body composition, glucose, and lipid status in rats during the 12-week study.

Probiotics and Isoflavones as a Promising Therapeutic for Calcium Status and Bone Health: A Narrative Review.

Probiotics have potential clinical effects for treating and preventing osteoporosis. Meanwhile, isoflavones have attracted much attention due to their ability to prevent postmenopausal symptoms. Research has established that probiotics and isoflavones can regulate hormones, immune cells, and the gastrointestinal system, acting as links in the gut-bone axis. However, combining the effects of probiotics and isoflavones on calcium status and bone health is a more novel and a still-evolving research area. Lactobacillus and Bifidobacterium are the foremost strains that influence bone health to a significant extent. Among the isoflavones, daidzein, genistein, and the metabolites of genistein (such as equol) stimulate bone formation. It can be concluded that probiotics and isoflavones promote bone health by regulating calcium uptake, gut microbiota, and various metabolic pathways that are associated with osteoblast activity and bone formation. Nevertheless, further experiments of probiotics and isoflavones are still necessary to confirm the association between calcium bioavailability and bone health.

The Impact of Multispecies Probiotics on Calcium and Magnesium Status in Healthy Male Rats.

Although probiotics have been discovered in numerous diseases in the last decade, there is little consensus on the relationship between probiotic properties and minerals balance and their distribution in the organism. This research aimed to evaluate the calcium (Ca) and magnesium (Mg) status in rats on a diet containing multispecies probiotics. Thirty male 10-week-old Wistar rats were selected and divided into three groups (n = 10 rats)-a group fed a standard diet (C), a group fed a low-dose of multispecies probiotics with 2.5 × 109 CFU per day (LD), and a group fed high-dose of multispecies probiotics 1 × 1010 CFU per day (HD) for 6 weeks. The results revealed that HD intake significantly increased the Ca concentration in hair and Mg concentration in femur bones. A significant positive correlation was found between calcium and magnesium levels in hair. The Ca/Mg molar ratio was lower in testicles in the groups with probiotics. In conclusion, multispecies probiotics altered the Ca concentration in hair and Mg level in femur bone, and also changed the molar ratio of these elements in testicles in male rats.

Gut microbiota profile in healthy Indonesians.

BACKGROUND: Recently, gut microbiota has been associated with various diseases other than intestinal disease. Thus, there has been rapid growth in the study of gut microbiota. Considering the numerous factors influencing gut microbiota such as age, diet, etc., area-based research is required. Indonesia has numerous different tribes and each of these tribes have different lifestyles. Hence, it is expected that each tribe has a specific gut microbiota. A deeper insight into the composition of gut microbiota can be used to determine the condition of gut microbiota in Indonesians and to consider which treatment may be suitable and effective to improve health status. AIM: To investigate the gut microbiota of Indonesian subjects represented by Javanese and Balinese tribes by analyzing fecal samples. METHODS: Fecal samples were collected from a total of 80 individuals with 20 in each of the young groups ranging from 25-45 years and the elderly group aged 70 years or more from two different regions, Yogyakarta and Bali. Fecal sample collection was performed at the end of the assessment period (day 14 ± 1 d) during which time the subjects were not allowed to consume probiotic or antibiotic products. The quantification of various Clostridium subgroups, Lactobacillus subgroups, Enterococcus, Streptococcus, Staphylococcus, Bacteroides fragilis group and Prevotella, Bifidobacterium and Atopobium cluster, Enterobacteriaceae and Pseudomonas was performed using the Yakult intestinal flora-scan (YIF-SCAN). RESULTS: The bacterial population in younger subjects' feces was higher than that in the elderly population, with a total of approximately 10.0 - 10.6 log10 bacterial cells/g feces. The most abundant bacteria in all groups were Clostridium, followed by Prevotella, Atopobium, Bifidobacterium and Bacteroides. In the elderly, an increase in Enterobacteriaceae, Coliform and Escherichia coli was found. In terms of bacterial counts in Yogyakarta, total bacteria, Clostridium coccoides (C. coccoides) group, Bifidobacterium, Prevotella, Lactobacillus plantarum subgroup, and Streptococcus were significantly higher (P < 0.05) in younger than elderly subjects, while the Lactobacillus gasseri subgroup, Lactobacillus casei subgroup, and Lactobacillus reuteri subgroup counts were significantly lower (P < 0.05) in younger subjects. In Balinese subjects, total bacteria, C. coccoides group, Clostridium leptum subgroup, Bacteroides fragilis group, and Prevotella were significantly higher (P < 0.05) in younger compared to elderly individuals, while the Lactobacillus ruminis subgroup, and Enterobacteriaceae were significantly lower (P < 0.05) in younger subjects. The results also revealed that, besides the C. coccoides group and Clostridium leptum group being the most abundant gut microbiota in both Yogyakarta and Balinese people, the latter was indicated by a higher Clostridium perfringens count, which was almost 10 times that of Yogyakarta subjects. This may be a response to different lifestyles in the different tribes; however, this phenomenon requires further extensive study. CONCLUSION: Bacterial populations were higher in younger than in elderly subjects. Most abundant bacterial groups were Clostridium, Prevotella, Atopobium, Bifidobacterium, and Bacteroides. The level of Clostridium perfringens in Yogyakarta subjects was lower than that in Balinese subjects.