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.

Soybean fermentation: Microbial ecology and starter culture technology.

Fermented soybean products, including Soya sauce, Tempeh, Miso, and Natto have been consumed for decades, mainly in Asian countries. Beans are processed using either solid-state fermentation, submerged fermentation, or a sequential of both methods. Traditional ways are still used to conduct the fermentation processes, which, depending on the fermented products, might take a few days or even years to complete. Diverse microorganisms were detected during fermentation in various processes with Bacillus species or filamentous fungi being the two main dominant functional groups. Microbial activities were essential to increase the bean's digestibility, nutritional value, and sensory quality, as well as lower its antinutritive factors. The scientific understanding of fermentation microbial communities, their enzymes, and their metabolic activities, however, still requires further development. The use of a starter culture is crucial, to control the fermentation process and ensure product consistency. A broad understanding of the spontaneous fermentation ecology, biochemistry, and the current starter culture technology is essential to facilitate further improvement and meet the needs of the current extending and sustainable economy. This review covers what is currently known about these aspects and reveals the limited available information, along with the possible directions for future starter culture design in soybean fermentation.

Development of Meat Substitutes from Filamentous Fungi Cultivated on Residual Water of Tempeh Factories.

In recent years, there has been an increased motivation to reduce meat consumption globally due to environmental and health concerns, which has driven the development of meat substitutes. Filamentous fungal biomass, commonly known as mycoprotein, is a potential meat substitute since it is nutritious and has filaments to mimic meat fibrils. The current study aimed to investigate the potential use of a cheap substrate derived from the food industry, i.e., residual water in a tempeh factory, for mycoprotein production. The type of residual water, nutrient supplementation, optimum conditions for biomass production, and characteristics of the mycoprotein were determined. The results showed that the residual water from the first boiling with yeast extract addition gave the highest mycoprotein content. The optimum growth condition was a pH of 4.5 and agitation of 125 rpm, and it resulted in 7.76 g/L biomass. The mycoprotein contains 19.44% (w/w) protein with a high crude fiber content of 8.51% (w/w) and a low fat content of 1.56% (w/w). In addition, the amino acid and fatty acid contents are dominated by glutamic acid and polyunsaturated fatty acids, which are associated with an umami taste and are considered healthier foods. The current work reveals that the residual boiling water from the tempeh factory can be used to produce high-quality mycoprotein.

Jack Bean (Canavalia ensiformis) Tempeh: ACE-Inhibitory Peptide Formation during Absorption in the Small Intestine.

Research background: High blood pressure is the most significant cause of mortality globally. Some fermented foods include ACE-inhibitory peptides that help fight this disease. The ability of fermented jack bean (tempeh) to inhibit ACE during consumption has not been demonstrated yet. This study identified and characterised ACE-inhibitory peptides from jack bean tempeh produced by small intestine absorption using the everted intestinal sac model. Experimental approach: Sequentially, the protein extract of jack bean tempeh and unfermented jack bean was hydrolysed using pepsin-pancreatin for 240 min. The hydrolysed samples were then evaluated for the peptide absorption using three-segmented everted intestinal sacs (duodenum, jejunum and ileum). The peptides absorbed from all intestinal segments were mixed in the small intestine. Results and conclusion: The data showed that both jack bean tempeh and unfermented jack bean had the same peptide absorption pattern, with the highest percentage of peptide absorption in the jejunum, followed by the duodenum and ileum. The absorbed peptides of jack bean tempeh exhibited equally strong activity of ACE inhibition in all intestinal segments, while the unfermented jack bean showed strong activity only in the jejunum. The mixture of the peptides from jack bean tempeh absorbed in the small intestine had higher ACE-inhibitory activity (81.09%) than the unfermented jack bean (72.22%). The peptides produced from jack bean tempeh were identified as pro-drug ACE inhibitors and had the mixed inhibition pattern. The mixture of peptides consisted of seven types of peptides with a molecular mass of 826.86-978.20 Da (DLGKAPIN, GKGRFVYG, PFMRWR, DKDHAEI, LAHLYEPS, KIKHPEVK, and LLRDTCK). Novelty and scientific contribution: This study discovered that consuming jack bean tempeh generated more potent ACE-inhibitory peptides during small intestine absorption than cooked jack beans. Absorbed tempeh peptides have high ACE-inhibitory activity.

Nuclear magnetic resonance spectroscopy and liquid chromatography-mass spectrometry metabolomics studies on non-organic soybeans versus organic soybeans (Glycine max), and their fermentation by Rhizopus oligosporus.

BACKGROUND: Soybeans (Glycine max) are high in proteins and isoflavones, which offer many health benefits. It has been suggested that the fermentation process enhances the nutrients in the soybeans. Organic foods are perceived as better than non-organic foods in terms of health benefits, yet little is known about the difference in the phytochemical content that distinguishes the quality of organic soybeans from non-organic soybeans. This study investigated the chemical profiles of non-organic (G, T, U, UB) and organic (C, COF, A, R, B, Z) soybeans (G. max [L.] Merr.) and their metabolite changes after fermentation with Rhizopus oligosporus. RESULTS: A clear separation was only observed between non-organic G and organic Z, which were then selected for further investigation in the fermentation of soybeans (GF and ZF). All four groups (G, Z, GF, ZF) were analyzed using nuclear magnetic resonance (NMR) spectroscopy along with liquid chromatography-tandem mass spectrometry (LC-MS/MS). In this way a total of 41 and 47 metabolites were identified respectively, with 12 in common. A clear variation (|log1.5 FC| > 2 and P < 0.05) was observed between Z and ZF: most of the sugars and isoflavone glycosides were found only in Z, while more amino acids and organic acids were found in ZF. An additional four metabolites clustered as C-glycosylflavonoids were discovered from MS/MS-based molecular networking. CONCLUSION: Chemical profiles of non-organic and organic soybeans exhibited no significant difference. However, the metabolite profile of the unfermented soybeans, which were higher in sugars, shifted to higher amino acid and organic acid content after fermentation, thereby potentially enhancing their nutritional value. © 2022 Society of Chemical Industry.

Microbiological quality of tempeh with different wraps: banana leaf versus plastic.

Tempeh is a traditional Indonesian fermented food widely consumed and became staple food in some Indonesian diet. Commercially, tempeh is available in banana leaf or plastic wraps. The wraps are not only important for fermentation and giving final form of tempeh,  but also potential source of food microbiome. This study aimed to investigate the effect of different packaging materials on the lactic acid bacteria (LAB) quantity and community composition of tempeh from three manufacturers with different production environments. Each tempeh wrapped with banana leaf or plastic during the fermentation process and then LAB from every tempeh sample were quantified using qPCR and plate count method. Terminal Restriction Fragment Length Polymorphism analysis was carried out to assign bacterial community composition from these samples. The LAB population quantity of fresh tempeh from three manufacturers were essentially similar around 10 log CFU/g. However, there is a difference in the proportion of cultured and uncultured bacteria. Principal Coordinate Analyses based on Bray-Curtis similarity matrices showed an apparent clustering pattern for tempeh samples according to tempeh manufacturers. Meanwhile, packaging materials did not significantly influence changes in  bacterial community composition. The result of this work could be used for determination, authentication, and improvement of tempeh quality.

Effect of Tempeh on Gut Microbiota and Anti-Stress Activity in Zebrafish.

Rhizopus oryzae is a fungus used to ferment tempeh in Indonesia and is generally recognized as safe (GRAS) for human consumption by the USA FDA. We previously assessed the effect of a tempeh extract on cortisol levels in zebrafish but did not include behavioral studies. Here, we measured the GABA content in three strains of Rhizopus oryzae, two isolated by us (MHU 001 and MHU 002) and one purchased. We then investigated the effect of tempeh on cortisol and the gut microbiota in a zebrafish experimental model. GABA concentration was the highest in MHU 002 (9.712 ± 0.404 g kg-1) followed by our MHU 001 strain and the purchased one. The fish were divided into one control group fed a normal diet and three experimental groups fed soybean tempeh fermented with one of the three strains of Rhizopus oryzae. After two weeks, individual fish were subjected to unpredicted chronic stress using the novel tank diving test and the tank light-dark test. Next-generation sequencing was used to analyze gut microbial communities and RT-PCR to analyze the expression of BDNF (brain-derived neurotrophic factor) gene and of other genes involved in serotonin signaling/metabolism in gut and brain. Tempeh-fed zebrafish exhibited increased exploratory behavior (less stress) in both tank tests. They also had significantly reduced gut Proteobacteria (include E. coli) (51.90% vs. 84.97%) and significantly increased gut Actinobacteria (include Bifidobacterium spp.) (1.80% vs. 0.79%). The content of Bifidobacteriumadolescentis, a "psychobiotic", increased ten-fold from 0.04% to 0.45%. Tempeh also increases BDNF levels in zebrafish brain. Rhizopus oryzae MHU 001 greatly improved the anti-stress effect of tempeh and microbiota composition in zebrafish gut.

Tempeh Consumption and Cognitive Improvement in Mild Cognitive Impairment.

INTRODUCTION: Tempeh consumption has been linked to the improvement of cognitive function in older people. However, to what extent the amount of microorganism or the size of tempeh serving consumed per day influences the benefit to cognitive functions has not yet been studied. METHODS: This experimental study involved a total of 90 respondents, who were divided into 3 groups: group A (consuming 100 g of Tempeh A/day), group B (consuming 100 g of Tempeh B/day), and group C (control). Intervention was given for 6 months. Cognitive assessments were done before and after the intervention. Blood uric acid level was checked at the end of intervention to examine the effect of tempeh consumption on this. The inclusion criteria were respondents aged 60 years or over with mild cognitive impairment (MCI) who agreed not to consume other fermented food during the study period. Respondents with diabetes were excluded. RESULTS: There were 84 subjects at the end of the study, majority being female (71.4%) and aged over 65 years (72.6%). An increase in global cognitive scores was found in both groups A and B. The increase in language domain scores was found only in group A. CONCLUSION: Both Tempeh A or Tempeh B consumption for 6 months appeared to be beneficial in improving global cognitive function of older people with MCI. Consuming Tempeh A, which had a lower number of microorganisms, was also associated with an improvement in the language domain.

Neuroprotective effect of tempeh against lipopolysaccharide-induced damage in BV-2 microglial cells.

Objectives: This study evaluated the bioactive composition of tempeh products and examined the effects of tempeh on BV-2 microglial cell cytotoxicity, neurotrophic effects, and expression of inflammatory genes.Methods: Tempeh products included soybean fermented by Rhizopus, soybean fermented through cocultivation with Rhizopus and Lactobacillus, and red bean fermented through cocultivation with Rhizopus and Lactobacillus (RT-C). We analyzed the bioactive contents of tempeh extracts and evaluated the effects of tempeh water extract on lipopolysaccharide (LPS)-treated BV-2 cells.Results: The results showed that RT-C water extract had the highest concentrations of γ-aminobutyric acid (GABA) and anthocyanin. The tempeh water extracts, especially RT-C, reduced the formation of LPS-induced reactive oxygen species, downregulated the levels of nitric oxide synthase and phospho-cyclic-AMP response element-binding protein, and upregulated the expression of brain-derived neurotrophic factor (BDNF).Discussion: Our data demonstrate that RT-C has the highest concentrations of GABA and anthocyanin, more effectively reduces oxidative stress and inflammation, and increases the expression of BDNF in LPS-induced BV-2 cells.

Enhanced vitamin B12 production in an innovative lupin tempeh is due to synergic effects of Rhizopus and Propionibacterium in cofermentation.

Fermentation represents a valuable and cost-effective approach for food stabilisation and nutritional improvement. Tempeh is an example of soybean solid-state fermentation. In this work, we investigated the possibility of producing a tempeh analogue containing high amounts of vitamin B12 using seeds of three different species of the legume lupin, namely Lupinus albus, L. angustifolius and L. mutabilis, with Rhizopus oligosporus and Propionibacterium freudenreichii cofermentation. Synergic effects of Rhizopus and Propionibacterium in increasing vitamin B12 up to 1230 ng/g dw was observed. These findings indicate that this cofermentation can improve lupin nutritional quality and safety to provide a tempeh analogue with added value for vegan and vegetarian communities and low-income populations. The level of potentially toxic lupin alkaloids was also monitored during the tempeh preparation.

Effect of controlled lactic acid fermentation on selected bioactive and nutritional parameters of tempeh obtained from unhulled common bean (Phaseolus vulgaris) seeds.

BACKGROUND: Tempeh is a traditional Indonesian food of high nutritional quality obtained by fungal fermentation of dehulled, soaked and cooked legumes. The aim of this research was to study the effect of Lactobacillus plantarum DSM 20174 activity on selected parameters of tempeh made from unhulled seeds of common bean (Phaseolus vulgaris). Lactobacillus plantarum cells were applied during soaking of seeds (submerged fermentation) or during solid state fermentation with Rhizopus microsporus var. chinensis (co-cultivation). RESULTS: Tempeh obtained from common beans contained 200 g kg⁻¹ protein of 34% in vitro bioavailability. Fungal fermentation caused decomposition of raffinose, stachyose and verbascose levels in seeds, on average by 93, 84 and 73% respectively. Enhanced antiradical (DPPH•, ABTS•+) capacity was accompanied by increased soluble phenol content. Application of Lactobacillus in the fermentation procedure increased tempeh protein and in vitro protein bioavailability by 18 and 17% respectively. Mixed culture tempeh contained lower levels of stachyose (25%), verbascose (64%) and condensed tannins (20%). Co-cultivation enhanced both DPPH•-scavenging activity and antioxidant capacity. CONCLUSION: The application of Lactobacillus in most cases improved the nutritional parameters of tempeh from unhulled common beans. It may also be recommended to obtain products with diverse antioxidant properties as compared with fungal fermentation alone.

Microbiological, sensory, and chemical properties of high-quality tempeh made with instant Mosaccha tempeh inoculum powder.

The combination of Saccharomyces cerevisiae and Rhizopus oligosporus liquid inoculum has been successfully used to ferment soybeans into tempeh that contains ß-glucan. However, using the liquid inoculum of these two microbes as a starter is impractical; so, developing an instant tempeh dry inoculum in powdered form, called the Mosaccha inoculum powder, for ease of use is necessary. This study aimed to determine the best concentration of instant Mosaccha inoculum powder to produce high-quality Mosaccha tempeh. The study used a Complete Randomized Block Design with seven different levels of instant Mosaccha inoculum powder percentage, ranging from 0.3% to 1.8% (w/w). A commercial tempeh inoculum, RAPRIMA, amounting to 0.2%, was used as control. Then, the microbiological (total mold and total yeast) and sensory (color, aroma, texture, and taste) properties were evaluated. The data obtained was analyzed statistically using analysis of variance (ANOVA) and Honestly Significant Difference (HSD) tests at the 5% level. The results showed that the percentage of instant Mosaccha inoculum powder significantly affected the microbiological and sensory properties of Mosaccha tempeh. A concentration of Mosaccha inoculum powder between 0.6% to 1.8% could produce good quality Mosaccha tempeh, but the best Mosaccha tempeh was produced with 1.5% instant Mosaccha inoculum powder, which met the Indonesian National Standards (SNI) 3144:2015, had a very favorable taste, and contained 0.49% ß-glucan. Therefore, Mosaccha inoculum in powdered form can be developed and used as a starter in making high-quality tempeh that contains ß-glucan.

Dextran and levan exopolysaccharides from tempeh-associated lactic acid bacteria with bioactivity against enterotoxigenic Escherichia coli (ETEC).

Soybean tempeh contains bioactive carbohydrate that can reduce the severity of diarrhea by inhibiting enterotoxigenic Escherichia coli (ETEC) adhesion to mammalian epithelial cells. Lactic acid bacteria (LAB) are known to be present abundantly in soybean tempeh. Some LAB species can produce exopolysaccharides (EPS) with anti-adhesion bioactivity against ETEC but there has been no report of anti-adhesion bioactive EPS from tempeh-associated LAB. We isolated EPS-producing LAB from tempeh-related sources, identified them, unambiguously elucidated their EPS structure and assessed the bioactivity of their EPS against ETEC. Pediococcus pentosaceus TL, Leuconostoc mesenteroides WA and L. mesenteroides WN produced both dextran (α-1,6 linked glucan; >1000 kDa) and levan (ß-2,6 linked fructan; 650-760 kDa) in varying amounts and Leuconostoc citreum TR produced gel-forming α-1,6-mixed linkage dextran (829 kDa). All four isolates produced EPS that could adhere to ETEC cells and inhibit auto-aggregation of ETEC. EPS-PpTL, EPS-LmWA and EPS-LmWN were more bioactive towards pig-associated ETEC K88 while EPS-LcTR was more bioactive against human-associated ETEC H10407. Our finding is the first to report on the bioactivity of dextran against ETEC. Tempeh is a promising source of LAB isolates that can produce bioactive EPS against ETEC adhesion and aggregation.

Soy-Based Tempeh as a Functional Food: Evidence for Human Health and Future Perspective.

Tempeh is an Indonesian traditional food made from fermented soybeans, which offers wide culinary use in East Asian countries. Similar to all fermented foods, its preparation offers the purpose of food preservation. However, preclinical studies have highlighted that microbial action leads to a modification in the nutritional composition of the food's matrix. Although there is a wide availability of data on the beneficial effect of soy, tempeh remains relatively unexplored, perhaps due to its limited diffusion in the world, which limits its research availability. However, available data suggest that tempeh may confer beneficial health effects due to the high bioavailability of nutrients and phytochemicals, showing ameliorative action on oxidative stress, glycaemic control, and blood lipid levels. Furthermore, the high biological value of tempeh means it can be used to optimize protein and caloric intake in athletes, vegetarians, and children. Moreover, the microbial fermentation used in the production of tempeh, in addition to improving the bioavailability of minerals, proteins, fibre, vitamins, and isoflavones, produces biopeptides whose biological effect is currently of great interest. Tempeh can be employed in traditional preparations as well as second-generation foods, such as plant-based meat substitutes, to provide functional and nutritional properties and a higher eco-friendly option compared to animal foods. This review aims to provide an overview of tempeh's properties, regarding human data and future research perspectives.

Mechanism of Action of Isoflavone Derived from Soy-Based Tempeh as an Antioxidant and Breast Cancer Inhibitor via Potential Upregulation of miR-7-5p: A Multimodal Analysis Integrating Pharmacoinformatics and Cellular Studies.

Breast cancer presents a significant global health challenge with rising incidence rates worldwide. Despite current efforts, it remains inadequately controlled. Functional foods, notably tempeh, have emerged as promising candidates for breast cancer prevention and treatment due to bioactive peptides and isoflavones exhibiting potential anticancer properties by serving as antioxidants, inducing apoptosis, and inhibiting cancer cell proliferation. This study integrates pharmacoinformatics and cellular investigations (i.e., a multifaceted approach) to elucidate the antioxidative and anti-breast cancer properties of tempeh-derived isoflavones. Methodologies encompass metabolomic profiling, in silico analysis, antioxidant assays, and in vitro experiments. Daidzein and genistein exhibited potential therapeutic options for breast cancer treatment and as antioxidant agents. In vitro studies also supported their efficacy against breast cancer and their ability to scavenge radicals, particularly in soy-based tempeh powder (SBT-P) and its isoflavone derivatives. Results have demonstrated a significant downregulation of breast cancer signaling proteins and increased expression of miR-7-5p, a microRNA with tumor-suppressive properties. Notably, the LD50 values of SBT-P and its derivatives on normal breast cell lines indicate their potential safety, with minimal cytotoxic effects on MCF-10A cells compared to control groups. The study underscores the favorable potential of SBT-P as a safe therapeutic option for breast cancer treatment, warranting further clinical exploration.

Gastroenteritis outbreak associated with unpasteurized tempeh, North Carolina, USA.

During an investigation of an outbreak of gastroenteritis caused by Salmonella enterica serovar Paratyphi B variant L(+) tartrate(+), we identified unpasteurized tempeh as a novel food vehicle and Rhizopus spp. starter culture as the source of the contamination. Safe handling of uncooked, unpasteurized tempeh should be emphasized for prevention of foodborne illnesses.

The effects of using Tempeh as a supplement for type 2 diabetes.

Studies suggest that the consumption of Tempeh can improve abnormal blood glucose and lipid parameters, although it remains still unclear as to whether Tempeh can improve tissue damage. In our study, db/db obese diabetic mice were given Tempeh 1 (300 mg/kg) and Tempeh 2 (600 mg/kg) for 3 months. The tissue samples collected were stained using different tissue-staining methodologies and were compared with the diabetic control group that was not given any Tempeh. Our results demonstrated that consuming high-dose Tempeh for 1 month could significantly reduce serum glucose and body weight in mice whereas the tissue section of our result could validate that consuming high-dose Tempeh for 3 months effectively improves lipid droplet size and lipid accumulation in the liver, aorta, and kidney of the mice. Moreover, an indication of the recovery of the damaged tissue could be observed in the heart and pancreatic tissue when high dosage of Tempeh was given as a treatment. Thus, it can be concluded that continuous consumption of Tempeh as a treatment could improve both blood glucose and body weight of diabetic mice while also improving lipid accumulation and tissue damage.

Physicochemical Properties of Yanggaeng with Added Tempeh Powder.

In this study, we investigated the physicochemical and antioxidative properties of the traditional Korean confectionery, Yanggaeng, when various amounts of tempeh powder (TP) were added. We replaced a portion of the white bean paste in Yanggaeng with TP at percentages of 0% (CON), 2% (TP2), 4% (TP4), and 6% (TP6) by total weight. The proximate composition results showed that TP6 exhibited the highest crude ash and crude protein contents, but its moisture content and carbohydrate content were the lowest compared to the CON. Tempeh addition altered the colorimetric properties by increasing the L* value, b* value, and browning index; however, tempeh addition did not alter the a* value. The results also showed that tempeh addition gradually decreased the pH of Yanggaeng. The Brix value was the highest in TP2; in TP4 and TP6, the Brix value gradually decreased, and these formulations exhibited lower Brix values than the CON. Furthermore, tempeh addition gradually induced antioxidative capacities, as evidenced by 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) radical scavenging activities. The results of this study demonstrate that the addition of tempeh to Yanggaeng alters its physicochemical properties and antioxidative capacity.

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.

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.