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14 pages, 3011 KiB  
Article
Ameliorative Effects of Soybean Powder Fermented by Bacillus subtilis on Constipation Induced by Loperamide in Rats
by Gi Soo Lee, Su Kang Kim, Ju Yeon Ban and Chung-Hun Oh
Int. J. Mol. Sci. 2025, 26(15), 7615; https://doi.org/10.3390/ijms26157615 - 6 Aug 2025
Abstract
Constipation is a prevalent gastrointestinal disorder that significantly impairs quality of life. While pharmacological agents such as loperamide are widely used to induce constipation in experimental models, there is increasing interest in natural alternatives for alleviating intestinal dysfunction. In this study, we investigated [...] Read more.
Constipation is a prevalent gastrointestinal disorder that significantly impairs quality of life. While pharmacological agents such as loperamide are widely used to induce constipation in experimental models, there is increasing interest in natural alternatives for alleviating intestinal dysfunction. In this study, we investigated the laxative effects of soybean powder fermented by Bacillus subtilis DKU_09 in a loperamide-induced rat model of constipation. The probiotic strain was isolated from cheonggukjang, a traditional Korean fermented soybean paste, and its identity was confirmed through 16S rRNA sequencing. Fermented soybean powder was characterized morphologically via scanning electron microscopy and chemically via HPLC to assess its isoflavone content. Rats were administered loperamide (5 mg/kg) for four days to induce constipation and were then treated with fermented soybean powder at doses of 100, 200, or 300 mg/kg. No pharmacological laxatives (e.g., PEG) were used as a positive control; instead, values from the treatment groups were compared with those from the loperamide-only constipation group. Key outcomes of fecal output, water content, colonic fecal retention, and gastrointestinal transit ratio were measured. The fermented product significantly improved stool frequency and moisture content, reduced colonic fecal retention, and restored gastrointestinal transit in a dose-dependent manner. Notably, the 300 mg/kg group demonstrated nearly complete recovery of fecal parameters without affecting body weight. Statistical analysis was performed using one-way ANOVA followed by Tukey’s post hoc test. These findings suggest that Bacillus subtilis-fermented soybean powder exerts synergistic laxative effects through the combined action of probiotic viability and fermentation-enhanced bioactive compounds such as aglycone isoflavones. This study supports the potential use of fermented soybean-based nutraceuticals as a natural and safe intervention for constipation and gastrointestinal dysregulation. Full article
(This article belongs to the Special Issue Functions and Applications of Natural Products)
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16 pages, 22496 KiB  
Article
Comparative Genomics and Adaptive Evolution of Bifidobacterium adolescentis in Geographically Distinct Human Gut Populations
by Pei Fu, Hao Qi and Wenjun Liu
Foods 2025, 14(15), 2747; https://doi.org/10.3390/foods14152747 - 6 Aug 2025
Abstract
Bifidobacterium adolescentis is prevalent in the gastrointestinal tract of healthy humans, and significantly influences host health. Recent studies have predominantly investigated the probiotic characteristics of individual strains and their specific metabolic roles, whereas analyses at the population genome level have been limited to [...] Read more.
Bifidobacterium adolescentis is prevalent in the gastrointestinal tract of healthy humans, and significantly influences host health. Recent studies have predominantly investigated the probiotic characteristics of individual strains and their specific metabolic roles, whereas analyses at the population genome level have been limited to date. This study conducted a comparative genomics analysis of 543 B. adolescentis genomes to explore genetic background variations and functional gene differences across geographically diverse populations. The results revealed significant differences in genome size and GC content among populations from Asia, Europe, and North America (p < 0.05). The pan-gene exhibited an open structure, reflecting the substantial genetic diversity within B. adolescentis. Functional annotation demonstrated that B. adolescentis possesses numerous protein-coding genes and abundant carbohydrate-active enzymes (CAZys) implicated in carbohydrate degradation and transformation. Population-specific CAZys were identified, suggesting adaptive evolution driven by distinct regional dietary patterns. Full article
(This article belongs to the Section Food Microbiology)
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16 pages, 2097 KiB  
Article
Dual Bioconversion Strategy: Synergistic Germination and Lactobacillus Fermentation Engineering for a γ-Aminobutyric Acid-Enriched Beverage from Brown Rice
by Di Yuan, Shan Zhang, Bin Hong, Shan Shan, Jingyi Zhang, Dixin Sha, Shiwei Gao, Qing Liu, Shuwen Lu and Chuanying Ren
Foods 2025, 14(15), 2733; https://doi.org/10.3390/foods14152733 - 5 Aug 2025
Abstract
Growing demand for plant-based nutraceuticals drives the need for innovative bioprocessing strategies. This study developed an integrated approach combining germination and Lactobacillus-mediated fermentation to produce a γ-aminobutyric acid (GABA)-enriched functional beverage from brown rice. Systematic screening identified an optimal rice cultivar for germination. [...] Read more.
Growing demand for plant-based nutraceuticals drives the need for innovative bioprocessing strategies. This study developed an integrated approach combining germination and Lactobacillus-mediated fermentation to produce a γ-aminobutyric acid (GABA)-enriched functional beverage from brown rice. Systematic screening identified an optimal rice cultivar for germination. Sequential enzymatic liquefaction and saccharification were optimized to generate a suitable hydrolysate. Screening of 13 probiotic strains revealed that a 10-strain Lactobacillus–Bifidobacterium consortium maximized GABA synthesis (12.2 mg/100 g). Fermentation parameters were optimized to 0.25% monosodium glutamate, 4% inoculum, 10 μmol/L pyridoxine hydrochloride, 37 °C, and 24 h. The resulting beverage achieved significantly elevated GABA concentrations while exhibiting low fat (0.2 g/100 g), reduced caloric content (233.6 kJ/100 g), and high viable probiotic counts (2 × 108 CFU/g). This strategy demonstrates significant potential for the scalable production of multifunctional, plant-based nutraceuticals with targeted bioactive components. Full article
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20 pages, 2168 KiB  
Article
Microbial Profiling of Buffalo Mozzarella Whey and Ricotta Exhausted Whey: Insights into Potential Probiotic Subdominant Strains
by Andrea Bonfanti, Romano Silvestri, Ettore Novellino, Gian Carlo Tenore, Elisabetta Schiano, Fortuna Iannuzzo, Massimo Reverberi, Luigi Faino, Marzia Beccaccioli, Francesca Sivori, Carlo Giuseppe Rizzello and Cristina Mazzoni
Microorganisms 2025, 13(8), 1804; https://doi.org/10.3390/microorganisms13081804 - 1 Aug 2025
Viewed by 111
Abstract
Buffalo mozzarella cheese whey (CW) and ricotta cheese exhausted whey (RCEW) are valuable by-products of the Mozzarella di Bufala Campana PDO production chain. This study characterized their microbial communities using an integrated culture-dependent and -independent approach. Metabarcoding analysis revealed that the dominance of [...] Read more.
Buffalo mozzarella cheese whey (CW) and ricotta cheese exhausted whey (RCEW) are valuable by-products of the Mozzarella di Bufala Campana PDO production chain. This study characterized their microbial communities using an integrated culture-dependent and -independent approach. Metabarcoding analysis revealed that the dominance of lactic acid bacteria (LAB), including Streptococcus thermophilus, Lactobacillus delbrueckii, and Lactobacillus helveticus, alongside diverse heat-resistant yeasts such as Cyberlindnera jadinii. Culture-based isolation identified subdominant lactic acid bacteria strains, not detected by sequencing, belonging to Leuconostoc mesenteroides, Enterococcus faecalis, and Enterococcus durans. These strains were further assessed for their probiotic potential. E. faecalis CW1 and E. durans RCEW2 showed tolerance to acidic pH, bile salts, and lysozyme, as well as a strong biofilm-forming capacity and antimicrobial activity against Bacillus cereus and Staphylococcus aureus. Moreover, bile salt resistance suggests potential functionality in cholesterol metabolism. These findings support the potential use of CW and RCEW as reservoirs of novel, autochthonous probiotic strains and underscore the value of regional dairy by-products in food biotechnology and gut health applications. Full article
(This article belongs to the Special Issue Microbial Fermentation, Food and Food Sustainability)
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10 pages, 1567 KiB  
Article
Correlation of Midgut Microbiota and Metabolic Syndrome-Related Lipids in Hemolymph Between Obese and Lean Silkworm Strains
by Huiduo Guo, Yalei Wang, Yu Guo, Xiangbiao Liu, Tao Gui, Mingfa Ling and Heying Qian
Insects 2025, 16(8), 798; https://doi.org/10.3390/insects16080798 - 1 Aug 2025
Viewed by 222
Abstract
Metabolic syndrome is a global health crisis. However, there are no effective therapeutic strategies for metabolic syndrome. Therefore, this study was conducted to find out a novel silkworm-based metabolic syndrome model that bridges microbial ecology and metabolic dysregulation by integrating hemolymph lipids and [...] Read more.
Metabolic syndrome is a global health crisis. However, there are no effective therapeutic strategies for metabolic syndrome. Therefore, this study was conducted to find out a novel silkworm-based metabolic syndrome model that bridges microbial ecology and metabolic dysregulation by integrating hemolymph lipids and midgut microbiota. Our results showed that the levels of HDL-C in the hemolymph of the lean silkworm strain were significantly higher than that in the obese silkworm strain. Furthermore, correlation analysis revealed that Lactococcus and Oceanobacillus were positively related to HDL-C levels, while SM1A02 and Pseudonocardia were negatively associated with HDL-C levels. These relationships between the identified bacteria in the midgut and HDL-C, known as the “good” lipid, in the hemolymph could help guide the development of new treatments for obesity and metabolic problems like high cholesterol in humans. Overall, our results not only established a framework for understanding microbiota-driven lipid dysregulation in silkworms but also offered potential probiotic targets and a bacterial biomarker for obesity and metabolic dysfunction intervention in humans. Full article
(This article belongs to the Section Insect Physiology, Reproduction and Development)
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15 pages, 1487 KiB  
Article
Protective Effects of a Bifidobacterium-Based Probiotic Mixture on Gut Inflammation and Barrier Function
by Yeji You, Tae-Rahk Kim, Minn Sohn, Dongmin Yoo and Jeseong Park
Microbiol. Res. 2025, 16(8), 168; https://doi.org/10.3390/microbiolres16080168 - 1 Aug 2025
Viewed by 329
Abstract
Disruption of the intestinal epithelial barrier is a key driver of gut-derived inflammation in various disorders, yet strategies to preserve or restore barrier integrity remain limited. To address this, we evaluated a four-strain Bifidobacterium mixture—selected for complementary anti-inflammatory potency and industrial scalability—in lipopolysaccharide [...] Read more.
Disruption of the intestinal epithelial barrier is a key driver of gut-derived inflammation in various disorders, yet strategies to preserve or restore barrier integrity remain limited. To address this, we evaluated a four-strain Bifidobacterium mixture—selected for complementary anti-inflammatory potency and industrial scalability—in lipopolysaccharide (LPS)-challenged RAW 264.7 macrophages and a Caco-2/THP-1 transwell co-culture model. Pretreatment with the probiotic blend reduced nitric oxide (NO) release in a dose-dependent manner by 25.9–48.3% and significantly down-regulated the pro-inflammatory markers in macrophages. In the co-culture system, the formulation decreased these markers, increased transepithelial electrical resistance (TEER) by up to 31% at 105 colony-forming unit (CFU)/mL after 48 h, and preserved the membrane localization of tight junction (TJ) proteins. Adhesion to Caco-2 cells (≈ 6%) matched that of the benchmark probiotic Lacticaseibacillus rhamnosus GG, suggesting direct epithelial engagement. These in vitro findings demonstrate that this probiotic mixture can attenuate LPS-driven inflammation and reinforce epithelial architecture, providing a mechanistic basis for its further evaluation in animal models and clinical studies of intestinal inflammatory disorders. Full article
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16 pages, 2259 KiB  
Article
Probiotic Potential and Characterization of Enterococcus faecium Strains Isolated from Camel Milk: Implications for Animal Health and Dairy Products
by Imen Fguiri, Manel Ziadi, Samira Arroum, Touhami Khorchani and Hammadi Mohamed
Fermentation 2025, 11(8), 444; https://doi.org/10.3390/fermentation11080444 - 31 Jul 2025
Viewed by 235
Abstract
In this study, 62 lactic acid bacteria (LAB) strains were isolated from raw camel milk and evaluated for their probiotic potential. The strains exhibited significant variability in their ability to withstand simulated gastrointestinal conditions. Of the isolates, only 26 survived exposure to pH [...] Read more.
In this study, 62 lactic acid bacteria (LAB) strains were isolated from raw camel milk and evaluated for their probiotic potential. The strains exhibited significant variability in their ability to withstand simulated gastrointestinal conditions. Of the isolates, only 26 survived exposure to pH 2, and just 10 were tolerant to 0.3% bile salts. Partial sequencing of the 16S rRNA gene identified all the strains as belonging to the species Enterococcus faecium. Several probiotic traits were assessed, including adhesion to gastric mucin and STC-1 intestinal epithelial cells, as well as auto-aggregation and co-aggregation capacities. Although adhesion to hydrophobic solvents such as chloroform and ethyl acetate was generally low to moderate, all the strains demonstrated strong adhesion to gastric mucin, exceeding 60% at all the growth stages. Notably, two strains—SCC1-33 and SLch6—showed particularly high adhesion to STC-1 cells, with values of 7.8 × 103 and 4.2 × 103 CFU/mL, respectively. The strains also exhibited promising aggregation properties, with auto-aggregation and co-aggregation ranging between 33.10% and 63.10%. Furthermore, all the isolates displayed antagonistic activity against Listeria innocua, Micrococcus luteus, and Escherichia coli. Cytotoxicity assays confirmed that none of the tested strains had harmful effects on STC-1 cells, indicating their safety and supporting their potential application as probiotics. Full article
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11 pages, 1139 KiB  
Article
Effect of Akkermansia muciniphila on GLP-1 and Insulin Secretion
by Ananta Prasad Arukha, Subhendu Nayak and Durga Madhab Swain
Nutrients 2025, 17(15), 2516; https://doi.org/10.3390/nu17152516 - 31 Jul 2025
Viewed by 299
Abstract
Background/Objectives: Gut microbiota research has gained momentum in recent years broadening knowledge of microbial components and their potential effects on health and well-being. Strong association between explicit microbes and metabolic diseases associated with obesity and type 2 diabetes mellitus, gastrointestinal disorders, neurodegenerative diseases, [...] Read more.
Background/Objectives: Gut microbiota research has gained momentum in recent years broadening knowledge of microbial components and their potential effects on health and well-being. Strong association between explicit microbes and metabolic diseases associated with obesity and type 2 diabetes mellitus, gastrointestinal disorders, neurodegenerative diseases, and even cancers have been established. Akkermansia muciniphila is a budding next-generation probiotic that plays an important role in systemic metabolism, intestinal health, and immune regulation, establishing strong implications for its use as a potent therapeutic intervention in diverse diseases. This project aimed at evaluating whether bacterial cell extracts of VH Akkermansia muciniphila (Vidya Strain; VS) can stimulate insulin secretion in INS-1 pancreatic beta cells and GLP-1 secretion in NCI-H716 human L-cells, both established in vitro models for studying metabolic regulation. Methods: Cultured VH Akkermansia muciniphila extracts were administered in a dose-dependent manner on INS-1 cells, and glucose-stimulated insulin secretion (GSIS) was measured via ELISA. Treated Human L-cell lines (NCI-H716) were analyzed for GLP-1 secretion. Results: Our study demonstrated that VH Akkermansia muciniphila extracts modestly increase insulin secretion from INS-1 beta cells and, more notably, induce a robust, dose-dependent rise in GLP-1 secretion from NCI-H716 L-cells, with the highest dose achieving over a 2000% increase comparable to glutamine. Conclusions: These findings suggest that VH A. muciniphila extracts may offer metabolic benefits by enhancing GLP-1 release, highlighting their potential for managing type 2 diabetes and obesity. Full article
(This article belongs to the Section Prebiotics and Probiotics)
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18 pages, 3259 KiB  
Article
Lactic Acid Bacteria in Chinese Sauerkraut: Its Isolation and In Vitro Probiotic Properties
by Ming-Yang Han, Wen-Yong Lou and Meng-Fan Li
Foods 2025, 14(15), 2690; https://doi.org/10.3390/foods14152690 - 30 Jul 2025
Viewed by 313
Abstract
Probiotics have been widely explored for their potential in managing hyperuricemia. However, their isolation and identification are fundamental prerequisites for practical application. In this study, 254 lactic acid bacteria (LAB) strains were isolated from Chinese sauerkraut and screened for probiotic potential based on [...] Read more.
Probiotics have been widely explored for their potential in managing hyperuricemia. However, their isolation and identification are fundamental prerequisites for practical application. In this study, 254 lactic acid bacteria (LAB) strains were isolated from Chinese sauerkraut and screened for probiotic potential based on genomic and phenotypic characteristics, as well as nucleoside-degrading activity relevant to decrease serum urate. Among them, Lactiplantibacillus plantarum (L. plantarum) F42 exhibited the highest bile salt tolerance (survivor rate: 19.46 ± 4.33%), strong adhesion to Caco-2 cells (1.89 ± 0.12%), effective nucleoside degradation (inosine: 5.46 ± 0.67 mg∙L−1∙min−1; guanosine: 3.84 ± 0.11 mg∙L−1∙min−1), and notable anti-listeria activity (inhibition zone: 6.9 ± 0.3 mm). Based on its functional profile, L. plantarum F42 was selected as a promising probiotic candidate for further investigation of its urate-lowering effects. This work provides a new insight into anti-hyperuricemia probiotic selection based on in vitro nucleoside-degrading activity. Full article
(This article belongs to the Section Food Biotechnology)
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18 pages, 2731 KiB  
Article
Dose-Dependent Anti-Inflammatory Effects of Live and Heat-Treated Ligilactobacillus salivarius and Bifidobacterium breve via NF-κB and COX-2 Modulation in an In Vitro Model of Airway Inflammation
by Marta Pagnini, Annalisa Visciglia, Giovanni Deusebio, Marco Pane, Alessandro Celi, Angela Amoruso and Tommaso Neri
Nutrients 2025, 17(15), 2504; https://doi.org/10.3390/nu17152504 - 30 Jul 2025
Viewed by 297
Abstract
Background: Probiotics are live microorganisms known for their health-promoting effects, particularly in modulating immune responses and reducing inflammation within the gastrointestinal tract. Emerging evidence suggests probiotics may also influence respiratory health, prompting investigation into their potential therapeutic application in lung inflammation. Methods: This [...] Read more.
Background: Probiotics are live microorganisms known for their health-promoting effects, particularly in modulating immune responses and reducing inflammation within the gastrointestinal tract. Emerging evidence suggests probiotics may also influence respiratory health, prompting investigation into their potential therapeutic application in lung inflammation. Methods: This study examined the anti-inflammatory effects of Ligilactobacillus salivarius (LS01 DSM 22775) and Bifidobacterium breve (B632 DSM 24706) on inflamed pulmonary epithelial cells. Lung carcinoma epithelial cells (A549) and normal bronchial epithelial cells (16HBE) were stimulated with IL-1β and treated with viable and heat-treated probiotics. Results: CCL-2 levels were significantly reduced by up to 40%, in A549 by viable form (105–107 AFU/g), instead of in 16HBE by heat-treated form (107–109 TFU/g). In A549 cells, TNF-α decreased by 20–80% with all formulations; instead, in 16HBE cells, IL-8 was reduced by viable strains (107 AFU/g) by approximately 50%, while heat-treated strains (109 TFU/g) decreased both IL-6 and IL-8 by 50%. All effective treatments completely inhibited IL-4 and eotaxin and suppressed NF-κB activation in both cell lines, with up to 80% reduction in phospho-p65 levels. In A549 cells, heat-treated strains fully blocked PGE2 production; instead, all four probiotics significantly inhibited COX-2 expression by approximately 50%. Conclusions: These findings demonstrate that both viable and heat-treated probiotics can modulate inflammatory responses in pulmonary epithelial cells, suggesting their potential application in inflammatory respiratory diseases. Heat-treated formulations may be particularly suited for local administration via inhalation, offering a promising strategy for targeting airway inflammation directly. Full article
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16 pages, 2615 KiB  
Article
The Prebiotic Potential of Porphyra-Derived Polysaccharides and Their Utilization by Lactic Acid Bacteria Fermentation
by Yu-Jyun Wei, Hong-Ting Victor Lin, Chorng-Liang Pan and Chung-Hsiung Huang
Fermentation 2025, 11(8), 435; https://doi.org/10.3390/fermentation11080435 - 29 Jul 2025
Viewed by 389
Abstract
Porphyra-derived polysaccharides (PPs) are promising prebiotic candidates due to their capacity to modulate gut microbiota and promote host health. However, their interactions with and utilization by probiotic microorganisms remain unclear. In this study, the fermentability of PPs by murine-derived lactic acid bacteria [...] Read more.
Porphyra-derived polysaccharides (PPs) are promising prebiotic candidates due to their capacity to modulate gut microbiota and promote host health. However, their interactions with and utilization by probiotic microorganisms remain unclear. In this study, the fermentability of PPs by murine-derived lactic acid bacteria (LAB) strains was investigated, with particular attention to strain-specific metabolic activity, carbohydrate utilization, and potential exopolysaccharide (EPS) production. All tested strains were capable of utilizing PPs to varying extents, with strain A10 exhibiting the highest level of carbohydrate consumption. Notably, strain A5 showed increased mannose concentrations following fermentation, suggesting the biosynthesis of mannose-rich EPSs. HPLC analysis confirmed the presence of high-molecular-weight polysaccharides ranging from 2.6 to 8.1 × 105 Da, indicative of EPS production. FT-IR spectroscopy further revealed spectral features consistent with EPS structures. The antibacterial activity of postbiotic compounds produced by LAB strains fermenting PPs against Escherichia coli and Staphylococcus aureus was observed. These findings demonstrate distinct metabolic adaptations of LAB strains to PPs and emphasize their potential as prebiotic substrates. Full article
(This article belongs to the Section Probiotic Strains and Fermentation)
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18 pages, 2062 KiB  
Review
Living Cultures in a Glass: The Health Promise of Probiotic Bacteria in Kombucha
by Tara Budimac, Aleksandra Ranitović, Olja Šovljanski, Dragoljub Cvetković and Ana Tomić
Fermentation 2025, 11(8), 434; https://doi.org/10.3390/fermentation11080434 - 29 Jul 2025
Viewed by 363
Abstract
Kombucha is a fermented tea beverage of Asian origin, widely consumed due to its functional properties; yet, it typically lacks sufficient levels of probiotic micro-organisms to be classified as a probiotic product. This review analyzes the occurrence of lactic acid bacteria (LAB) in [...] Read more.
Kombucha is a fermented tea beverage of Asian origin, widely consumed due to its functional properties; yet, it typically lacks sufficient levels of probiotic micro-organisms to be classified as a probiotic product. This review analyzes the occurrence of lactic acid bacteria (LAB) in kombucha, reporting that concentrations rarely exceed 4–5 log CFU/mL and often decline during fermentation or storage. Strategies to enhance probiotic viability, including the use of robust LAB strains and encapsulation technologies, are critically evaluated. Notably, encapsulation using pea and whey protein has been shown to sustain LAB levels above 6 log CFU/mL during fermentation and up to 21 days under refrigerated storage for whey protein. Fortified kombucha beverages with probiotic strains have also been shown to possess enhanced functional and health-promoting benefits compared to traditional control samples. Despite promising approaches, inconsistencies in microbial survival and regulatory constraints remain key challenges. Future research should focus on the optimization of delivery systems for probiotic cultures, identification of kombucha-compatible LAB strains and standardized protocols to validate probiotic efficacy in real-world beverage conditions. Full article
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22 pages, 2147 KiB  
Article
Streamlining Bacillus Strain Selection Against Listeria monocytogenes Using a Fluorescence-Based Infection Assay Integrated into a Multi-Tiered Validation Pipeline
by Blanca Lorente-Torres, Pablo Castañera, Helena Á. Ferrero, Sergio Fernández-Martínez, Suleiman Adejoh Ocholi, Jesús Llano-Verdeja, Farzaneh Javadimarand, Yaiza Carnicero-Mayo, Amanda Herrero-González, Alba Puente-Sanz, Irene Sainz Machín, Isabel Karola Voigt, Silvia Guerrero Villanueva, Álvaro López García, Eva Martín Gómez, James C. Ogbonna, José M. Gonzalo-Orden, Jesús F. Aparicio, Luis M. Mateos, Álvaro Mourenza and Michal Letekadd Show full author list remove Hide full author list
Antibiotics 2025, 14(8), 765; https://doi.org/10.3390/antibiotics14080765 - 29 Jul 2025
Viewed by 275
Abstract
Background/Objectives: Listeria monocytogenes is a foodborne pathogen of major public health concern due to its ability to invade host cells and cause severe illness. This study aimed to develop and validate a multi-tiered screening pipeline to identify Bacillus strains with probiotic potential [...] Read more.
Background/Objectives: Listeria monocytogenes is a foodborne pathogen of major public health concern due to its ability to invade host cells and cause severe illness. This study aimed to develop and validate a multi-tiered screening pipeline to identify Bacillus strains with probiotic potential against L. monocytogenes. Methods: A total of 26 Bacillus isolates were screened for antimicrobial activity, gastrointestinal resilience, and host cell adhesion. A fluorescence-based infection assay using mCherry-expressing HCT 116 cells was used to assess cytoprotection against L. monocytogenes NCTC 7973. Eight strains significantly improved host cell viability and were validated by quantification of intracellular CFU. Two top candidates were tested in a murine model of listeriosis. The genome of the lead strain was sequenced to evaluate safety and biosynthetic potential. Results: B. subtilis CECT 8266 completely inhibited intracellular replication of L. monocytogenes in HCT 116 cells, reducing bacterial recovery to undetectable levels. In vivo, it decreased splenic bacterial burden by approximately 6-fold. Genomic analysis revealed eight bacteriocin biosynthetic clusters and silent antibiotic resistance genes within predicted genomic islands, as determined by CARD and Alien Hunter analysis. The strain also demonstrated bile and acid tolerance, as well as strong adhesion to epithelial cells. Conclusions: The proposed pipeline enables efficient identification of probiotic Bacillus strains with intracellular protective activity. B. subtilis CECT 8266 is a promising candidate for translational applications in food safety or health due to its efficacy, resilience, and safety profile. Full article
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21 pages, 3912 KiB  
Article
Screening and Phenotyping of Lactic Acid Bacteria in Boza
by Xudong Zhao, Longying Pei, Xinqi Wang, Mingming Luo, Sihan Hou, Xingqian Ye, Wei Liu and Yuting Zhou
Microorganisms 2025, 13(8), 1767; https://doi.org/10.3390/microorganisms13081767 - 29 Jul 2025
Viewed by 325
Abstract
The aim of this study was to isolate and identify lactic acid bacteria (LAB) from a traditional fermented beverage, Boza, and to conduct an in-depth study on their fermentation and probiotic properties. The fermentation (acid production rate, acid tolerance, salt tolerance, amino acid [...] Read more.
The aim of this study was to isolate and identify lactic acid bacteria (LAB) from a traditional fermented beverage, Boza, and to conduct an in-depth study on their fermentation and probiotic properties. The fermentation (acid production rate, acid tolerance, salt tolerance, amino acid decarboxylase activity) and probiotic properties (gastrointestinal tolerance, bile salt tolerance, hydrophobicity, self-aggregation, drug resistance, bacteriostatic properties) of the 16 isolated LAB were systematically analyzed by morphological, physiological, and biochemical tests and 16S rDNA molecular biology. This analysis utilized principal component analysis (PCA) to comprehensively evaluate the biological properties of the strains. The identified LAB included Limosilactobacillus fermentum (9 strains), Levilactobacillus brevis (2 strains), Lacticaseibacillus paracasei (2 strains), and Lactobacillus helveticus (3 strains). These strains showed strong environmental adaptation at different pH (3.5) and temperature (45 °C), with different gastrointestinal colonization, tolerance, and antioxidant properties. All the strains did not show hemolytic activity and were inhibitory to Staphylococcus aureus, and showed resistance to kanamycin, gentamicin, vancomycin, and streptomycin. Based on the integrated scoring of biological properties by principal component analysis, Limosilactobacillus fermentum S4 and S6 and Levilactobacillus brevis S5 had excellent fermentation properties and tolerance and could be used as potential functional microbial resources. Full article
(This article belongs to the Special Issue Microbial Fermentation in Food Processing)
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15 pages, 2248 KiB  
Article
Effects of Treadmill Exercise on Gut Microbiota in Alzheimer’s Disease Model Mice and Wild-Type Mice
by Zhe Zhao, Xingqing Wu, Wenfeng Liu, Lan Zheng and Changfa Tang
Microorganisms 2025, 13(8), 1765; https://doi.org/10.3390/microorganisms13081765 - 29 Jul 2025
Viewed by 291
Abstract
There is a growing body of research showing that Alzheimer’s disease (AD) is related to enteric dysbacteriosis. Exercise can be effective in alleviating AD, but the effects that exercise has on the gut microbiota in AD patients needs to be further studied. Through [...] Read more.
There is a growing body of research showing that Alzheimer’s disease (AD) is related to enteric dysbacteriosis. Exercise can be effective in alleviating AD, but the effects that exercise has on the gut microbiota in AD patients needs to be further studied. Through this study, we aimed to investigate the differences in the diversity of gut microorganisms between AD model mice and wild-type mice and the effect that treadmill exercise has on the composition of the gut microbiota in both types of mice. C57BL/6 wild-type mice were randomly divided into a sedentary control group (WTC) and an exercise group (WTE); APP/PS1 double transgenic mice were also randomly divided into a sedentary control group (ADC) and an exercise group (ADE). After the control group remained sedentary for 12 weeks and a 12-week treadmill exercise intervention was adopted for the exercise group, the rectal contents were collected so that they could undergo V3-V4 16S rDNA sequencing, and a comparative analysis of the microbial composition and diversity was also performed. The alpha diversity of the gut microbiota in AD mice was lower than that in wild-type mice, but exercise increased the gut microbial diversity in both types of mice. At the phylum level, the dominant microorganisms in all four groups of mice were Bacteroidetes and Firmicutes. There was an increase in the Bacteroidetes phylum in AD mice. Treadmill exercise reduced the abundance of Bacteroidetes in both groups of mice, whereas the abundance of Firmicutes increased. At the genus level, Muribaculaceae, the Lachnospiraceae_NK4A136_group, Alloprevotella, and Alistipes were in relatively high abundance. Muribaculaceae and Alloprevotella were in greater abundance in AD mice than in wild-type mice, but both decreased after treadmill exercise. Through performing linear discriminant analysis effect size (LEfSe), we found that the dominant strains in AD mice were Campilobacterota, Helicobacteraceae, Escherichia–Shigella, and other malignant bacteria, whereas exercise resulted in an increase in probiotics among the dominant strains in both types of mice. Although gut microbial diversity decreases and malignant bacteria increase in AD mice, treadmill exercise can increase gut microbial diversity and lead to the development of dominant strains of probiotics in both types of mice. These findings provide a basis for applying exercise as a treatment for AD. Full article
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