The interest in fermented food products has been increasing in recent years. Details about their microbial composition and the effects of their consumption on the human gut microbiome are of particular interest. However, evidence regarding their potential to increase gut microbial diversity, a measure likely associated with health, is lacking. To address this, we analyzed the microbial composition of commercially available fermented vegetables using 16S rRNA sequencing. We also conducted a pilot study to assess the feasibility of studying the effects of regular consumption of fermented vegetables on the gut microbiome. Six healthy male volunteers participated in a randomized crossover trial, with two two-week intervention phases. Volunteers consumed 150 g/d of either sauerkraut or a variety of six different commercially available fermented vegetables. This study is registered at the German Clinical Trials Register (DRKS-ID: DRKS00014840). Lactobacillales was the dominant family in all fermented vegetables studied. However, the alpha diversity, richness and evenness of the microbiota differed substantially among the different products. The number of species per product varied between 20 and 95. After consumption of both sauerkraut and the selection of fermented vegetables, we observed a slight increase in alpha diversity. Specifically, the amount of the genus Prevotella decreased while the amount of Bacteroides increased after both interventions. However, these initial observations need to be confirmed in larger studies. This pilot study demonstrates the feasibility of this type of research. Full article

Aspergillus-derived cellulase, which is involved in the production of Aspergillus-fermented foods, has been employed in the food and animal feed industries. However, the effect of dietary Aspergillus cellulase on health is poorly understood. Previously, we discovered that supplemental Aspergillus-derived protease and lipase preparations had substantial bifidogenic effects on rats fed a high-fat diet. Therefore, this study reports on the effects of a 0.1% dietary Aspergillus-derived cellulase preparation (CEL) on the gut microbiota of rats fed a high-fat diet. Gene sequencing analysis of 16S rRNA revealed that CEL treatment markedly affected the microbiota profiles of the cecal contents (p < 0.05). Notably, CEL markedly increased the relative abundance (RA) of typical probiotics, such as Bifidobacterium and Lactobacillus, at the genus level (28- and 5-fold, respectively, p < 0.05). Similarly, at the family level, CEL treatment significantly increased the RA of Bifidobacteriaceae and Lactobacillaceae (p < 0.05). Furthermore, CEL increased the RA of other genera, such as Collinsella and Enterococcus, but decreased the RA of Oscillospira, Dorea and Coprobacillus (p < 0.05). The effects on these genera are similar to those reported for typical prebiotic oligosaccharides. Overall, this study demonstrates the prebiotic-like effects of dietary CEL by significantly increasing Bifidobacterium and Lactobacillus abundance. Full article

In this study, the antioxidant mechanism of Astragalus membranaceus fermented by Lactiplantibacillusplantarum MG5276 (MG5276F-AM) was evaluated in HepG2 cells and in an animal model. HPLC analysis was performed to confirm the bioconversion of the bioactive compounds in A. membranaceus by fermentation. Calycosin and formononetin, which were not detected before fermentation (NF-AM), were detected after fermentation (MG5276F-AM), and its glycoside was not observed in MG5276F-AM. In HepG2 cells, MG5276F-AM alleviated H₂O₂-induced oxidative stress by mediating lipid peroxidation and glutathione levels, and upregulated antioxidant enzymes including catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GPx). In the tBHP-injected mouse model, administration of MG5276F-AM reduced hepatic aspartate transaminase, alanine transaminase, and lipid peroxidation. MG5276F-AM also modulated antioxidant enzymes as well as HepG2 cells. Thus, fermentation of A. membranaceus with L. plantarum MG5276 elevated the isoflavonoid aglycone by hydrolysis of its glycosides, and this bioconversion enhanced antioxidant activity both in vitro and in vivo. Full article

Aspergillus-derived protease and lipase, which are involved in the production of Aspergillus-fermented foods, are consumed as digestive enzyme supplements. A marked bifidogenic effect of supplemental Aspergillus protease preparation (AP) in rats fed with a high-fat diet was identified. This study was conducted to examine whether the consumption of Aspergillus-derived lipase exerts similar bifidogenic effect. Rats were fed diets supplemented with either an Aspergillus-derived lipase preparation (AL) or AP at 0.1% for two weeks. 16S rRNA gene sequencing analysis indicated that supplemental AL and AP markedly influenced cecal microbial community. At the phylum level, treatment with AL and AP resulted in a lower relative abundance of Firmicutes and Bacteroidetes, but a higher relative abundance of Actinobacteria and Proteobacteria than the control rats (p < 0.05). At the genus level, AL and AP remarkedly elevated the relative abundances of Bifidobacterium, Collinsella, and Enterococcus, but significantly reduced those of Oscillospira, Dorea, and Coprobacillus (p < 0.05). These modulations were similar to those reported by several studies with typical prebiotic oligosaccharides. Notably, the bifidogenic effect of AL was much greater than that of AP. Our results show that the two different Aspergillus-derived preparations, AL and AP, have strong bifidogenic effects and can change the microbiota’s composition. Full article

Lećevački cheese is a traditional Croatian hard cheese, which belongs to the group of hard Mediterranean cheeses produced from mixed milk (cow and sheep). The aim of this research was to determine the influence of different ratios and compositions of sheep milk on the composition and yield of Lećevački cheese. A total of 15 batches of Lećevački cheese were selected containing different ratios of sheep and cow milk from the regular production of a dairy plant. The ratio of sheep milk was as follows: up to 39%, from 40 to 44%, and from 45 to 50%. For each ratio, five batches were randomly selected. A higher ratio of sheep milk caused a noticeable increase in fat, protein, lactose, and total solids content, while the content of solids-not-fat significantly (p < 0.05) increased. A similar trend was found for casein content (p < 0.1). The highest ratio of sheep milk in mixed milk increased (p < 0.05) the protein content by almost 1%. However, the results showed that it is not reasonable to increase the sheep milk ratio in mixed milk above 44% (v/v) because it causes a higher (p < 0.01) moisture content in the cheese, as well as a lower fat content (p < 0.01) and fat recovery (p = 0.07) during the manufacturing of Lećevački cheese. Full article

In this study, we aim to decrease the fermentation time and to produce low-fat set-type kefir with adequate textural properties using microbial transglutaminase without inactivation. In addition, we reveal the effect of microbial transglutaminase, during and after fermentation, on kefir made with cow, goat, or donkey milk, which is a novel approach. Fermentation is followed by continuous pH and viscosity measurements; the final product is characterized by dry matter content, whey separation, protein pattern, and viscosity parameters, as well as gel firmness. The results show that already 0.5 U/g protein dosage of MTGase decreases pH levels independent of milk type, but MTGase does not influence the kinetics of fermentation. Apparent viscosity could be measured from different stages of fermentation depending on milk type (cow milk, 6 h; goat milk, 8 h; and donkey milk, 9 h). The final product characteristics show that the higher the casein ratio of the applied milk, the better the viscosity and gel firmness of the kefir due to the high reaction affinity of MTGase. Full article

Miang, a traditional fermented tea from Northern Thailand, potentially hosts beneficial probiotic bacteria. A total of 133 isolates of lactic acid bacteria (LAB) isolated from Miang were evaluated for probiotic potential. Among them, 5 strains showed high tolerance to bile and acidic conditions and were selected for further evaluation. All selected strains showed inhibitory activity against human pathogens, including Bacillus cereus, Staphylococcus aureus, and Salmonella ser. Typhimurium. Nucleotide sequences analysis of the 16S rRNA gene revealed that 3 isolates were identified as Lactobacillus pentosus; the remaining were L. plantarum and Pediococcus pentosaceus, respectively. All 5 strains showed a high survival rate of more than 90% when exposed to simulated gastrointestinal conditions and were also susceptible to antibiotics such as erythromycin, tetracycline, and gentamycin, and resistant to vancomycin, streptomycin, and polymycin. In addition, the selected isolates exhibited different degrees of cell surface hydrophobicity (58.3–92.9%) and auto-aggregation (38.9–46.0%). The antioxidant activity reflected in DPPH scavenging activities of viable cells and their cell-free culture supernatants (CFCS) were also found in selected LAB isolates. Moreover, selected LAB isolates showed ability to grow on commercial prebiotics (GOS, FOS or XOS). The preliminary study of spray-drying using cyclodextrin as thermoprotectant suggested that all strains can be designed as a powdered formulation. L. pentosus A14-6 was the best strain, with high tolerance against simulated gastrointestinal conditions, high cell surface hydrophobicity, effective response to tested commercial oligosaccharides, especially XOS, and the highest cell antioxidant properties. L. pentosus A14-6 was therefore targeted for further applications in food and synbiotic applications. Full article

Osteoporosis is the most common bone disease associated with low bone mineral density. It is the process of bone loss and is most commonly caused by decreased estrogen production in women, particularly after menopause. Pueraria lobata, which contains various metabolites, especially isoflavone, is widely known as regulator for bone mineral contents. In this study, the effects of the P. lobata extract (PE) with or without fermentation with Lactobacillus paracasei JS1 (FPE) on osteoporosis were investigated in vitro and in vivo. The effects of PE and FPE on human osteoblastic MG63 cells, RAW 264.7 cells, and ovariectomized (OVX)-induced model mice were analyzed at various ratios. We found that FPE increased calcium deposition and inhibited bone resorption by in vitro assay. Furthermore, treatment with PE and FPE has significantly restored destroyed trabecular bone in the OVX-induced bone loss mouse model. Overall, FPE demonstrated bioactivity to prevent bone loss by decreasing bone turnover. Full article

Traditional fermented foods have been recognized by various communities to be good for health since ancient times. There is a provincial legacy of traditional fermented foods among the ethnic population of North-East India. Fermented bamboo shoots (local name: Tuaither), soybeans (Bekang), and pork fat (Sa-um) are famous in the Mizoram state and represent a primary portion of the daily diet. These foods are prepared using methods based on cultural traditions inherited from previous generations, and prepared using a relatively uncontrolled fermentation process. Analysis of the bacterial diversity in these foods can provide important information regarding the flavor and texture of the final products of fermentation. Unfortunately, studies on the microbial composition and health benefits of such traditional fermented foods have rarely been documented. Therefore, the present study aims to highlight this bacterial diversity, along with the proximate composition of different traditional fermented foods (Tuaither, Bekang and Sa-um) primarily consumed in Mizoram state, India. Samples were collected on three different days of fermentation (3rd, 5th and 7th day), and bacterial diversity analysis was performed using the V3-V4 variable region of 16S rRNA gene with Illumina sequencing. Results revealed differences in the bacterial composition of dominant group members among all of the three food types. Firmicutes (82.72–94.00%), followed by Proteobacteria (4.67–15.01%), were found to dominate to varying degrees in all three of the fermented foods. However, at genus level high variation was observed in bacterial composition among these three different types of fermented foods. Lactobacillus (91.64–77.16%), Staphylococcus (52.00–17.90%), and Clostridium (72.48–55.40%) exhibited the highest relative abundances in the Tuaither, Bekang and Sa-um foods, respectively, in descending order from the 3rd to 7th day of fermentation. A few of the bacterial genera such as Lactobacilli were positively correlated with fermented bamboo shoot samples, and Staphylococcus was positively correlated with protein, carbohydrate and crude fiber content in soybean samples. In general, Tuaither, Bekang and Sa-um exhibited distinct differences in bacterial composition. This variation may be due to differences in the raw materials and/or methods used in the preparation of the different fermented food products. This is the first study to describe the bacterial composition of these traditional fermented foods using high-throughput sequencing techniques, and could help to drive research attention to comprehensive studies on improving understanding of the role of microbial communities in the preparation of traditional foods and their health benefits. Full article

The microalga Chlorella and strains of Bifidobacterium have been used in human or animal food supplements for decades because of their positive health effects. The presented study assessed different properties of C. vulgaris and its combination with bifidobacteria with the aim to develop new functional foods. The growth of four bifidobacteria strains in milk and whey supplemented with 1.0% (w/v) C. vulgaris and the immunomodulatory effects of aqueous Chlorella solutions (0.5%, 1.0%, and 3.0%) on human peripheral mononuclear cells were evaluated. Furthermore, synergistic effects on lipid metabolism of rats fed a high-fat diet with Chlorella and B. animalis subsp. lactis BB-12^® were analysed. Chlorella had a positive growth-promoting effect on the tested bifidobacteria (p < 0.05), and significantly increased the secretion of inflammatory cytokines (tumor necrosis factor-α, interleukin-10, and interleukin-6), depending on the concentration of Chlorella (p < 0.05). After 8 weeks, significant synergistic effects of Chlorella and bifidobacteria on triglyceride levels in rat heart, liver, and serum were observed (p < 0.05). These results demonstrate that various combinations of Chlorella and bifidobacteria have significant potential for the development of new fermented products, dependent on the algal species, probiotic strain, application form, and concentrations for acceptable sensory quality for consumers. Full article

This article presents new data on Bifidobacterium longum MC-42—a strain that has been actively used for the preparation of commercial dairy products in Russia for almost 40 years. It was demonstrated that this strain possesses high activities of β-galactosidase, α-glucosidase, and leucine arylaminidase; inhibits the growth of pathogens such as Salmonella typhimurium, Staphylococcus aureus, and Escherichia coli; and can efficiently remove cholesterol from the cultural medium. The resistance of B. longum MC-42 determined for 15 commonly used antibiotics was in agreement with those previously reported for Bifidobacterium spp. The absence of frequently transmittable antibiotic resistance genes in the genome and the lack of undesirable activity of β-glucuronidase proved the safe use of B. longum MC-42 as a probiotic and starter culture. Additionally, the impact of two growth-promoting additives—yeast extract or milk protein hydrolysate containing supplementation—on the B. longum MC-42 fermentation profile was assessed. The introduction of these additives increases the maximum attainable viable cell count by orders of magnitude, significantly changed the profile of aminopeptidase activities in extracellular extracts, and influenced the antioxidant and antihypertensive properties of the obtained fermented products. Full article

In Taiwan, adult hyperuricemia affects as many as 1 in 4 males and 1 in 6 females, who are predominantly young adults aged 19–45. In this study, lactic acid bacteria (LAB) with acid tolerance, bile salt tolerance and high affinity to intestinal cells were extracted from the side products of alcohol fermentation (distillers’ grains). These bacteria were evaluated for their ability to lower uric acid levels. Qualitative identification and quantitative analysis were performed using high-performance liquid chromatography (HPLC) on the purine-degrading enzymes to select purine-decomposing LAB for animal testing. When the final concentration of purine compounds reached 0.1% and 1%, seven strains of LAB showed potential in degrading purine compounds. HPLC was used to analyze their purine-degrading abilities, and the three best performing LAB strains, (107) 8–16, (107) tau 1–3, and (107) 6–10 were screened for further animal testing with Wistar rats. By the third week, the results showed that strain (107) 6–10 could prevent formation and reduce the levels of blood urea nitrogen (BUN) in yeast extract/potassium oxonate-induced hyperuricemia. The multi-strain lactic acid bacteria (MLAB) performed best for uric acid reduction in the serum and down regulated BUN. Yeast extract/potassium oxonate-induced hyperuricemia had no impact on serum creatinine, while LAB did not affect the creatinine concentration. In summary, MLAB not only protects kidney function but is also effective in regulating uric acid concentration in the body. Hence, MLAB can be used as a functional food supplement that prevents or aids the treatment of hyperuricemia in a rodent model. Full article

Bacillus velezensis ST03 and ST32, Bacillus amyloliquefaciens ST06 and ST109, and Bacillus subtilis ST08 were isolated from artisanal-produced kimchi and were identified based on 16S rRNA partial sequencing. DNA obtained from the investigated bacilli generated positive results for lichenicidin, iturin, subtilosin, and surfactin on a strain-specific basis. The strains were found to produce antimicrobial metabolites with activity levels ranging between 800 and 1600 AU/mL on a strain-specific basis, as determined against Listeria monocytogenes ATCC15313. Moreover, all tested strains in this study were still active after treatment with proteolytic enzymes, even with reduced inhibition zones compared to the controls, pointing to additional antimicrobial activity possibly related to a non-proteinaceous molecular structure. Most probably these strains may express surfactin as an additional factor in their complex antimicrobial activity. B. amyloliquefaciens ST09 and B. velezensis ST03 and ST32 were characterized as positive for β-hemolysis. B. subtilis ST08 was shown to be positive for hblC and nheC and B. amyloliquefaciens ST109 for nheB. B. amyloliquefaciens ST109 generated positive results for gelatinase activity. The ability of the studied Bacillus strains to metabolize different carbohydrate sources was done based on the API50CHB test, while the enzyme production profile was recorded by the APIZym kit. All studied strains were positive producers for biogenic amines production. Studied Bacillus spp. strains were resistant to some of the evaluated antibiotics, tested according to recommendations of CLSI and EFSA. Full article

Although results of recent studies suggest that fermented foods strongly affect the gut microbiota composition and that they relieve inflammatory bowel disease symptoms, some reports have described that fermented foods increase some inflammation markers based on differences in fermented food materials. This study evaluated the effects of fermented plant extract (FPE) on dextran sulfate sodium (DSS)-induced colitis in mice and the effects on fecal microbiota composition in humans. Mice fed 5% FPE with 3% DSS (FPE group) showed no body weight loss, atrophy of colonic length, or bloody stool, similar to mice fed a basal diet (negative group), whereas mice fed 3% DSS (positive group) exhibited those effects. Concentrations of inflammation markers IL-6 and TNF-α were not significantly different between FPE and negative groups; however, those concentrations became higher in the positive group. 16S ribosomal RNA gene sequencing was used to characterize fecal microbiota in healthy women before and after 3-month FPE supplementation. The FPE supplementation induced increases in Firmicutes phyla and in Clostridiales order, which play a central role in inflammation suppression. These results suggest that FPE enhances Clostridiales growth in the gut and that it has an anti-inflammatory effect. Full article

Although high-fat and high-salt diets are considered risk factors for hypertension, the intake of salty soybean-based fermented foods has beneficial effects. This study explored the potential of Chinese traditional fermented soy sauce (CTFSS) in preventing hypertension by analyzing its effects on adipogenesis and the renin-angiotensin-aldosterone system (RAAS). Male Sprague-Dawley (SD) rats were divided into four groups (n = 6): normal diet (ND), high-fat diet (HD), high-fat diet with saline (HDS, NaCl-8%), and high-fat diet with Chinese traditional soy sauce (HDCTS, NaCl-8%). Each group is administrated 12 weeks by oral gavage as 10 mL/kg dose, respectively. CTFSS supplementation resulted in significantly lower body weight, epididymal fat weight, and systolic blood pressure. Additionally, it decreased the serum total cholesterol (TC), triglyceride (TG), alanine aminotransferase (ALT), aspartate aminotransferase (AST), renin, angiotensin II (Ang II), angiotensin-converting enzyme (ACE), and aldosterone levels. It also increased the urinary volume and improved sodium and potassium ion balance. The gene levels showed significant enhancements in the mRNA levels of renin-angiotensin-aldosterone system-related and adipogenesis-related genes. In addition, CTFSS may prevent hypertension-associated kidney injury. Therefore, this study demonstrates that CTFSS has no harmful effects on hypertension. In contrast, the beneficial effects of CTFSS intake in ameliorating hypertension were shown. Full article

Alzheimer’s disease (AD) is a common neurodegenerative disease characterized by continuous accumulation of β-amyloid (Aβ) in the brain. Deep ocean water (DOW) with rich inorganic salts and minerals was proven to promote fungi growth and metabolism. Cordyceps cicada, a functional food fungus, can produce higher anti-oxidant and anti-inflammatory compounds including adenosine, polysaccharide, and N(6)-(2-Hydroxyethyl) adenosine (HEA). This study used DOW as the culture water of C. cicadae NTTU 868 for producing DOW-cultured C. cicadae (DCC), and further investigated the effects and mechanisms on improving the memory deficit and repressing risk factors expressions in Aβ40 and streptozotocin (STZ)-induced Alzheimer’s disease rats model. In the results, DCC including mycelium and filtrate had adenosine, HEA, polysaccharide, and intracellular Mg²⁺ after fermentation with DOW. DCC had more effect on the improvement of memory deficit because it suppressed Aβ40 and streptozotocin (STZ) infusion caused BACE, pro-inflammatory factors expressions, and Aβ40 accumulation by increasing sRAGE expression in the brain. Furthermore, DCC enhanced the MAGT1 expression due to high organic magnesium, which can reverse Aβ40-induced cortex magnesium deficiency and further repress Aβ40 accumulation. Full article

The control of the immune system of pigs after weaning is important in pig farming because productivity depends on the survival of the post-weaned pigs. Previously, antibiotics would have been administered in the case of infectious diseases to increase the survival rate of post-weaned pigs, but now, the use of antibiotics is strictly restricted in order to prevent other problems such as the occurrence of antibiotic-resistant pathogens. In this study, the effect of fermented kefir as a functional feed additive as a replacement to antibiotics was evaluated in terms of the microbial profile in fecal samples, immunological factors in the blood of pigs, growth performance measured as average daily gain (ADG) and the feed conversion rate (FCR) of post-weaned pigs. In the kefir-treated group, the number of lactic acid bacteria and Bacillus spp. in the fecal samples of the pigs increased with the kefir treatments. Interestingly, the number of coliform groups as opportunistic pathogens was reduced in the fecal samples of pigs treated with kefir. We found out that treatment with kefir enhanced the innate immunity of post-weaned pigs though the reduction of IL-6 as a proinflammatory cytokine and an increase in IgG as an immunoglobulin, enhancing immunological defense against pathogens. Finally, after treatment with kefir, we observed that the ADG of post-weaned pigs increased to 135.6% but FCR decreased to 92.2%. Therefore, this study shows that fermented kefir can be used as a functional feed additive and an antibiotic alternative in order to improve both the innate immune system and growth performance of post-weaned pigs. Full article

Litopenaeus vannamei, known as whiteleg shrimp, is susceptible to infection by pathogenic microorganisms such as viruses and bacteria. Therefore, the prevention of infections in this shrimp is important to regulate the outbreaks of pathogenic microorganisms. In this study, we investigated the effects of kefir as a functional feed additive on innate immunity, survival against WSSV (White Spot Syndrome Virus) and productivity of L. vannamei. As a result, the treatment of kefir could upregulate six of seven genes crucial for innate immunity of L. vannamei. Also, the treatment of kefir directly improved the survival rate of L. vannamei against WSSV infection. Finally, in order to determine whether kefir can improve the productivity of shrimp, we carried out field tests in three aquaculture farms in South Korea. The weight of shrimp fed kefir was increased by 120% as well as the length, compared with that of the control group. These results demonstrate that kefir can be utilized as a functional feed additive to improve both innate immunity and productivity of L. vannamei in shrimp farming with no use of antibiotics. Full article

Probiotics and synbiotics are known to have beneficial effects on human health and disease. Hirsutism, a disorder that is characterised by the presence of coarse terminal hairs in a male-like pattern, is usually caused by elevated androgen levels in blood plasma. This disorder is usually observed in PCOS women and it is linked to insulin resistance (IR). Although idiopathic hirsutism (IH) is not shown to have excess androgen production from the ovarian and adrenal glands, increased 5α-reductase in peripheral tissues and insulin resistance are common observations. The effect of probiotics and synbiotics have been recently studied on PCOS women; androgens were also included in the hormonal groups that were investigated. Only a few studies focus on hirsutism and the potential effect of the beneficial microbes mentioned, whereas the increasing interest on insulin resistance and synbiotics indicate a potential beneficial effect on hirsutism through the management of insulin resistance. Full article