Search Results (260)

Fermented products have recently garnered substantial interest in both research and commercial contexts. Although probiotic fermentation is predominantly practiced with dairy, fruits, vegetables, and grains, its application to dual-purpose food-medicine materials like Ganoderma lucidum has been comparatively underexplored. In this study, Ganoderma lucidum fermented juice (GFJ) served as the substrate and was fermented with five probiotic strains. The optimal inoculation ratios—determined by employing a uniform design experiment—were as follows: Bifidobacterium animalis 6.05%, Lacticaseibacillus paracasei 9.52%, Lacticaseibacillus rhamnosus 6.63%, Pediococcus pentosaceus 21.38%, and Pediococcus acidilactici 56.42%. Optimal fermentation parameters established by response surface methodology included 24 h of fermentation at 37 °C, a final cell density of 5 × 10⁶ CFU/mL, and a sugar content of 4.5 °Brix. Experiments with RAW264.7 macrophages revealed that GFJ significantly promoted both phagocytic activity and nitric oxide (NO) secretion, indicating enhanced immune characteristics as a result of fermentation. Untargeted metabolomics profiling of GFJ across different fermentation stages showed upregulation of functional metabolites, including polyphenols, prebiotics, functional oligosaccharides, and Ganoderma triterpenoids (GTs)—notably myricetin-3-O-rhamnoside, luteolin-7-O-glucuronide, raffinose, sesamose, and Ganoderma acids. These increments in metabolic compounds strongly correlate with improved functional properties in GFJ, specifically heightened superoxide dismutase activity and immunomodulatory capacity. These results highlight an effective approach for developing functionally enriched fermented products from medicinal fungi, with promising applications in functional food and nutraceutical industries. Full article

Fortifying sourdough bread with functional ingredients is an effective strategy to enhance nutritional value, bioactive content, and sensory quality. The novel strain Lactiplantibacillus paracasei SP5 was incorporated into mother sponges together with cereal-based derivatives, such as trahanas or delignified wheat bran (2% and 5% w/w), and their effects on sourdough bread functionality were compared with a control bread without supplementation. Sourdough bread containing 5% w/w trahanas exhibited the highest lactic acid bacteria (10.4 log cfu/g), approximately 22% higher than the control, and the strongest fermentation activity, leading to acetic acid levels 27% higher than the control. This sourdough bread sample also had 73% higher total phenolic content (112.1 mg GAE/100 g) and approximately 27% higher antioxidant activity compared to control bread. In addition, phytic acid degradation exceeded 91%, representing a 16% higher reduction than the control, thereby improving mineral bioavailability. Shelf-life was also markedly extended, in terms of mould and rope spoilage compared with the control. Wheat bran-supplemented breads showed moderate improvements, with LAB counts, antioxidant activity, and phytic acid reduction about 10–25% higher than the control. Overall, these findings demonstrate that L. paracasei SP5 in combination with nutrient-rich, low-cost cereal-based derivatives can significantly enhance the technological, nutritional, and bioactive properties of sourdough breads. This approach provides a simple, industrially feasible strategy for producing functional breads with improved health-promoting and preservative attributes. Full article

With growing emphasis on antibiotic-free poultry production, functional probiotics represent a promising strategy to improve gut health and reduce pathogen transmission. This study characterized three lactic acid bacteria (LAB) strains Lactobacillus delbrueckii subsp. bulgaricus NRRL-B-548 (LD), Lacticaseibacillus paracasei DUP-13076 (LP), and Lacticaseibacillus rhamnosus NRRL-B-442 (LR) for their probiotic potential and evaluated their efficacy against Salmonella enterica in poultry. The LAB strains were assessed for acid and bile tolerance, lysozyme resistance, cholesterol assimilation, antimicrobial activity, surface hydrophobicity, epithelial adherence, hemolysis, and antibiotic susceptibility. Genomic analysis was performed to identify genes associated with probiotic functionality. The protective potential of LR and LP was further validated in hatchlings using a hatchery spray model challenged with Salmonella Enteritidis. All strains survived simulated gastric and intestinal conditions, exhibited strong adhesion to epithelial cells, and demonstrated high hydrophobicity, indicating robust colonization capacity. The LAB significantly inhibited Salmonella Enteritidis, S. Typhimurium, and S. Heidelberg growth in vitro and remained sensitive to clinically relevant antibiotics. In vivo application of LR and LP to hatching eggs markedly reduced S. Enteritidis colonization in the liver, spleen, and ceca of hatchlings. Further, genomic profiling of the LAB strains revealed genes for bacteriocin production, exopolysaccharide synthesis, and carbohydrate metabolism supporting probiotic function. In summary, the evaluated LAB strains exhibit multiple probiotic attributes and strong anti-Salmonella activity, confirming their potential as safe, hatchery-applied probiotics for improving gut health and biosecurity in poultry production systems. Full article

Background/Objectives: Impaired intestinal barrier function is a hallmark of inflammatory bowel disease (IBD). Akkermansia muciniphila and its outer membrane protein Amuc_1100 can enhance this barrier, but the clinical application of Amuc_1100 is limited by the fastidious growth of its native host. This study aimed to overcome this by utilizing the robust probiotic Lacticaseibacillus paracasei L9 for targeted Amuc_1100 delivery. Methods: We engineered Lc. paracasei L9 to express Amuc_1100 via intracellular (pA-L9), secretory (pUA-L9), and surface-display (pUPA-L9) strategies. Their efficacy was assessed in Lipopolysaccharide (LPS)-induced macrophages and a dextran sulfate sodium (DSS)-induced colitis mouse model, evaluating inflammation, barrier integrity, and mucosal repair. Results: The secretory (pUA-L9) and surface-display (pUPA-L9) strains most effectively suppressed pro-inflammatory cytokines (IL-6, IL-1β, and TNF-α) in macrophages. In mice, both strains alleviated colitis and outperformed native A. muciniphila in improving disease activity. Crucially, they exhibited distinct, specialized functions: pUA-L9 acted as a systemic immunomodulator, reducing pro-inflammatory cytokines (IL-6, IL-1β, and TNF-α), elevating anti-inflammatory mediators (IL-4 and IL-10), and promoting goblet cell differentiation; notably, the inhibitory effect of pUA-L9 on IL-6 expression was approximately 2-fold greater than that of pUPA-L9. In contrast, pUPA-L9 excelled in local barrier repair, uniquely restoring mucus layer integrity (Muc1, Muc2, and Tff3) and reinforcing tight junctions (ZO-1, Occludin, Claudin1, Claudin3, and Claudin4). In particular, pUPA-L9 increased Muc2 expression by approximately 3.6-fold compared with pUA-L9. Conclusions: We demonstrate that the subcellular localization of Amuc_1100 within an engineered probiotic dictates its therapeutic mode of action. The complementary effects of secretory and surface-displayed Amuc_1100 offer a novel, spatially targeted strategy for precision microbiome therapy in IBD. Full article

Inoculated fermentation can enhance the flavor, nutrition, and functionality of juice. The lactic acid bacteria (LAB) are commonly used as starter cultures. This study screened LAB for orange juice fermentation and optimized fermentation factors using response surface methodology (RSM) to improve GABA content in orange juice. A total of 52 LAB strains were screened, and Lacticaseibacillus paracasei ZY (Lcb. paracasei ZY) and Lacticaseibacillus rhamnosus SN12 (Lcb. rhamnosus SN12) presented higher GABA yields and adaptability to substrates. The optimized fermentation factors for GABA enhancement in orange juice were as follows: initial pH of 5.5, fermentation temperature of 37 °C, soluble solids content of 12.0 °Bx, inoculum ratio of Lcb. paracasei ZY to Lcb. rhamnosus SN12 as 1:1, inoculum size of 6 Log CFU/mL, and fermentation time of 96 h. Under these optimized conditions, the GABA content reached 0.89 g/L, representing a 39.06% increase compared to uninoculated orange juice. This indicates that RSM-based optimization is conducive to increasing GABA content in orange juice and provides a scientific basis for the development of GABA-enriched functional fermented juices. Full article

In our previous work, we reported for the first time the presence of enterocin-encoding genes in novel Lacticaseibacillus paracasei (L. paracasei) and Levilactobacillus brevis (Lev. brevis) strains isolated from artisanal dairy products made from raw cow milk. The aim of this study was to isolate enterocin-positive lactic acid bacteria (LAB) from artisanal dairy products and assess their technological characteristics and safety for potential application in food systems. LAB isolates were characterized using phenotypic tests, Matrix-Assisted Laser Desorption/Ionization Time-of-Flight (MALDI-TOF) identification, and PCR detection of enterocin genes, followed by evaluation of their physiological and technological properties and a comprehensive safety assessment, including antimicrobial resistance, virulence, biogenic amine, and integron genes. Two strains, L. paracasei S2 and Lev. brevis S62, carried enterocin genes (entAS-48 and entQ) and exhibited strong acidifying and proteolytic activities, along with antibacterial effects against foodborne pathogens and reference strains. Both isolates tolerated environmental stresses, including low pH, and lacked virulence factors, clinically relevant antibiotic resistance genes, biogenic amine production, and integron elements. These results indicate that the strains are safe, multifunctional, and suitable for developing regionally adapted dairy products, highlighting artisanal dairy products as a valuable source of novel LAB with promising biotechnological applications. Full article

Second cheese whey (SCW), a major by-product of ricotta cheese production, poses significant environmental challenges due to its high organic load. Biohydrogen (bio-H₂) and poly-β-hydroxybutyrate (PHB) production offer a sustainable reuse of SCW, that provides ideal nutrients for microbial growth. This study aimed to convert SCW into Bio-H₂ and PHB using a 5-liter tubular bioreactor in a sequential lactic fermentation and photofermentation system. Two lighting conditions were tested: white LED (WL) and selected LED (SL). Optimal results were achieved with a co-inoculum of Lactococcus lactis MK L84 and Lacticaseibacillus paracasei MK L49 at pH 4.5–5.5, followed by photofermentation with Rhodopseudomonas palustris 42OL under SL condition. The process yielded an average of 0.47 L of H₂ per liter of substrate and 1.66% wPHB/wCDW. This approach successfully transformed dairy waste into high-value products, promoting circular economy principles. Full article

Background: This study investigated the phenotypic and genotypic antibiotic resistance profiles of 50 Lactic Acid Bacteria (LAB) strains—25 Lactiplantibacillus plantarum and 25 Lacticaseibacillus paracasei—isolated from traditional Sardinian fermented foods of animal origin. Methods: The sensitivity of the isolates to antibiotics such as β-lactams, tetracyclines, aminoglycosides, macrolides, phenicols, and glycopeptides was initially assessed using disc diffusion and minimum inhibitory concentration (MIC) tests. Subsequently, PCR analyses were performed on both genomic DNA and plasmid DNA to detect blaZ, tet(W), strA, aac(6′)-Ie–aph(2″)-Ia, and vanX genes associated with resistance to ampicillin, tetracycline, streptomycin, gentamicin, and vancomycin. Results: The analysis revealed that L. plantarum strains frequently carried the tet(W) gene on the chromosome and strA on plasmids, while vanX was detected in most strains as a chromosomal determinant. By contrast, L. paracasei strains exhibited a predominantly plasmid-mediated distribution of resistance genes. For example, strA, aac(6′)-Ie–aph(2″)-Ia and blaZ were often found on plasmids, whereas vanX remained chromosomally encoded. Phenotypic assays confirmed high intrinsic resistance to vancomycin in both species, with L. plantarum showing a higher overall frequency and diversity of resistant phenotypes compared to L. paracasei. Conclusions: The co-occurrence of multiple resistance determinants, including plasmid-encoded ones, in most strains suggests that even autochthonous isolates from artisanal products may represent potential reservoirs for transmissible resistance genes. Full article

The influence of lactic acid bacteria (LAB) strains of various species isolated from Chinese traditional sourdough on the properties of rice sourdough and the textural and flavor qualities of steamed rice bread (SRB) was investigated. Lactiplantibacillus plantarum-fermented rice sourdough had a higher total titratable acidity (13.10 mL) than the other groups. Strains Lacticaseibacillus paracasei PC1 (LPC), Lactobacillus helveticus H1 (LH), Lactobacillus crustorum C1 (LC), Lactobacillus paralimentarius PA1 (LPA), and Lactiplantibacillus plantarum P1 (LP) showed marked protein hydrolysis during rice sourdough fermentation and increased free amino acid levels in rice sourdoughs relative to the control. The Fourier Transform Infrared Spectroscopy results indicated that LAB fermentation could promote the strengthening of inter-intramolecular hydrogen bonds and cause modifications in protein structures; however, these effects varied among the different strains. The LC and LPC strains had the most significant effect on improving the specific volume and textural properties of SRBs. Gas chromatography-mass spectrometry (GC-MS) and GC-ion mobility spectrometry (IMS) identified 33 and 35 volatile compounds, respectively, in the LAB-fermented SRBs, and differentiation was observed in the volatile profiles of SRBs made using different LAB strains. The differential impacts of LAB strains during rice sourdough fermentation can assist in the selection of candidate microorganisms for the production of high-quality gluten-free rice products. Full article

Chronic psychological stress impairs neuroendocrine balance and increases the risk of mental health disturbances, including anxiety, sleep disruption, and low mood. The gut–brain axis has emerged as a promising target for stress modulation, particularly through psychobiotic interventions. This randomized, double-blind, placebo-controlled trial evaluated the effects of a combined psychobiotic formulation (Neuralli Mood), comprising live Lactiplantibacillus plantarum PS128 (PS128) and heat-treated Lacticaseibacillus paracasei PS23 (HT-PS23), on the psychological and physiological stress responses in a high-stress occupational population. A total of 116 healthy participants with elevated perceived stress (PSS ≥ 14), primarily firefighters, were randomly assigned to receive the dual-strain supplement or placebo for 8 weeks. Stress-related outcomes were assessed by using validated psychological scales and serum biomarkers. Compared with placebo, the psychobiotics group showed significantly greater reductions in overall job stress perception (JSS), state anxiety (STAI), and insomnia severity (ISI) (all p < 0.05). Additionally, serum adrenocorticotropic hormone (ACTH) and norepinephrine levels were significantly reduced post-intervention, whereas cortisol levels remained unchanged. These findings suggested that combining live and heat-treated psychobiotic strains may provide a safe and effective strategy for alleviating psychological stress and regulating neuroendocrine function in high-risk populations. Full article

Acne vulgaris is a common chronic inflammatory skin condition. Conventional acne treatments are often limited by adverse effects, driving interest in alternative therapies. This study explored the multifunctional bioactivities of a lyophilized cell-free supernatant (LCFS) derived from Lacticaseibacillus paracasei T0901, isolated from fermented palm sap, with a focus on its antimicrobial, antibiofilm, and antimelanogenic potential for dermatological applications. Antimicrobial activity was evaluated using agar well diffusion and broth microdilution assays against acne-associated pathogens, while antibiofilm effects were quantified via crystal violet staining. Antimelanogenic activity was assessed in α-melanocyte-stimulating hormone (α-MSH)-stimulated B16F10 melanoma cells by measuring melanin content and tyrosinase activity. Whole-genome sequencing was performed to identify genes linked to observed bioactivities, and molecular docking was used to predict metabolite–protein interactions. The LCFS exhibited strong inhibitory activity against acne-associated bacteria, with inhibition zones of C. acnes (10.67 ± 0.58 mm), S. epidermidis (21.00 ± 0.00 mm), and S. aureus (20.00 ± 0.00 mm), and a minimum inhibitory concentration of 25 mg/mL. Biofilm formation was significantly reduced by 62.98 ± 3.54%. In α-MSH-stimulated B16F10 cells, LCFS treatment (10 mg/mL) significantly decreased melanin content (73.23 ± 2.36%) and intracellular tyrosinase activity (68.19 ± 6.29%) relative to control. Genomic analysis revealed antioxidant-related genes (sodA, trxB, nox), pigmentation regulators (mco, fcbD), and buk (butyrate kinase), supporting the observed bioactivities. Molecular docking further demonstrated strong binding affinities of LCFS-derived metabolites to tyrosinase and MITF, suggesting modulation of melanogenic pathways. Collectively, these results indicate that L. paracasei T0901 produces safe postbiotic compounds with potent antimicrobial, antibiofilm, and antimelanogenic activities, highlighting its promise as a multifunctional ingredient in probiotic-based skincare formulations. Full article

Donkey milk is an underexplored biological niche with distinctive nutritional and microbiological properties, potentially harboring lactic acid bacteria (LAB) with technological or probiotic value. In this study, raw milk from the endangered Zamorano-Leonesa donkey breed was stored at 4 °C for 24 h to simulate realistic cold-chain conditions and favor the recovery of cold-tolerant microorganisms. Fourteen isolates were obtained, eight of which belonged to LAB or species with potential technological interest and were selected for functional evaluation. Phenotypic screening showed that most isolates tolerated acidic conditions (pH 2.5) and that four also resisted 0.3% bile salts. Acidification assays in pasteurized donkey milk revealed variable fermentation performance, with L. mesenteroides subsp. mesenteroides B8 and Lacticaseibacillus paracasei subsp. tolerans B19 displaying the most favorable profiles. These two strains were selected for genome sequencing. Genomic analysis revealed genes associated with acid and bile resistance, adhesion, cold and environmental stress responses, and carbohydrate metabolism. Both genomes also encoded biosynthetic gene clusters linked to secondary metabolites, including β-lactones, lincosamides, and RiPP-like compounds. No acquired antimicrobial resistance genes were detected. To our knowledge, this is the first study combining isolation, phenotypic screening, and genome-based characterization of cold-tolerant LAB from Zamorano-Leonesa donkey milk. Our findings highlight this milk as a valuable reservoir of safe, cold-adapted microorganisms with promising applications in functional dairy products and food biotechnology. Full article

Previous research showed that three strains of lactic acid bacteria (LAB)—Leuconostoc mesenteroides, Lacticaseibacillus paracasei and Loigolactobacillus coryniformis—exhibited in vitro anti-listerial activity. The objectives of the present study was: (1) to determine whether the initial pH of heat-treated reconstituted milk affects the growth of Listeria monocytogenes, in monoculture and coculture with each of the LAB strains; and (2) to compare the capacity of the LAB strains to inhibit L. monocytogenes in the milk model. Monoculture and coculture challenge tests were conducted in milk adjusted to three initial pH levels of 5.5, 6.0, and 6.5. A pH-driven model and a Jameson-effect model were fitted to the growth curves. The former provided more precise estimates than the latter. In monoculture, L. mesenteroides exhibited the fastest development at all initial pH levels whereas, in coculture, this strain more effectively controlled L. monocytogenes by reducing its growth rates. As the initial milk pH increased, the maximum concentration of L. monocytogenes in monoculture and in coculture also increased, although the LAB strains were all able to reduce the pathogen’s maximum concentration. These findings demonstrated that adjusting milk to a more acidic pH before fermentation provides an additional barrier against the development of L. monocytogenes. Full article

Elderly individuals are more vulnerable to disease due to their increased frailty. Emerging evidence highlights the potential of probiotics as geroprotective agents by maintaining gut health and modulating key physiological processes involved in aging, such as inflammation, cognitive functions, and metabolism. Here, we investigated the geroprotective potential of four probiotic strains (Lacticaseibacillus paracasei LPC1114, Limosilactobacillus reuteri PBS072, Bifidobacterium breve BB077, and Bifidobacterium animalis subsp. lactis BL050) using Caenorhabditis elegans as an aging model. Mid-life healthspan parameters were assessed, including lifespan, motility, ROS levels, lipofuscin accumulation, and cognitive capabilities. The probiotics exhibited strain-specific effects. L. reuteri PBS072 and B. lactis BL050 significantly increased locomotion by 20% and decreased ROS levels by 70% and 30% respectively, suggesting enhanced oxidative stress response and neuromuscular maintenance. B. breve BB077, L. paracasei LPC1114, and L. reuteri PBS072 enhanced associative learning performance, whereas B. lactis BL050 improved chemotactic response. Notably, only L. paracasei LPC1114 and L. reuteri PBS072 extended the maximum lifespan by 4 and 5 days, respectively, an effect mediated by the longevity-related genes skn1, sir2.1, and daf16. Our findings highlight the multifaceted, strain-specific geroprotective properties of probiotics and support their potential as microbiome-based interventions to promote healthy aging. Full article

Industrial hemp (Cannabis sativa L.; Cannabaceae), traditionally cultivated for fiber, also represents a valuable source of nutrient-rich seed oil. In this study, cold-pressed hemp seed oils from three Romanian varieties (Teodora, Silvana, and Armanca) were evaluated for their fatty acid composition, minor bioactive constituents, antioxidant activity, growth-promoting property toward probiotic strains in vitro, and molecular docking interactions with probiotic targets. Gas chromatography revealed a fatty acid profile dominated by linoleic (49.4–51.9%), oleic (16.3–22.8%), and α-linolenic acids (9.8–14.4%), resulting in favorable PUFA/SFA ratios (5.17–6.39) and ω-6/ω-3 ratios (3.93–5.53). The oils also contained phenolics (118–160 mg GAE/kg), chlorophylls (6.18–8.31 mg/kg), and carotenoids (2.58–3.37 mg/kg), which contributed to their antioxidant activity (DPPH inhibition 35.92 µM TE/100 g–43.37 µM TE/100 g). Broth microdilution assays against Lacticaseibacillus rhamnosus GG, L. paracasei ATCC BAA-52, and L. acidophilus ATCC 4356 demonstrated strain- and dose-dependent potential to promote probiotic growth under in vitro conditions. While L. rhamnosus and L. paracasei were inhibited at low concentrations and only mildly stimulated at higher levels, L. acidophilus showed robust growth promotion, reaching +54.7% effect and CP = 1.55 with Teodora oil at 16 mg/mL. Molecular docking highlighted strong binding affinities of γ-linolenic and linoleic acids with key metabolic enzymes involved in probiotic metabolism (hydratase, enolase, glyceraldehyde-3-phosphate dehydrogenase, ribonucleoside hydrolase), forming stable hydrophilic and hydrophobic interactions which are explored in defining the stability of the ligand-protein complexes. These results indicate that both major fatty acids and minor bioactive constituents contribute to the nutritional and antioxidant value of Romanian hemp seed oils and reveal a potential to promote probiotic growth under in vitro conditions, as supported by complementary in silico evidence. Full article