Efficacy of combination of Lacticaseibacillus rhamnosus and Bifidobacterium bifidum probiotic strains against mixed infections of bovine Rotavirus A and Cryptosporidium parvum induced diarrhea by modulating host immune response in mouse model.

Obesity rates are rising globally, placing a significant strain on individuals, society, and economies. The gut microbiota (GM) plays a pivotal role in the development of obesity. Many studies have identified differences in GM composition between obese and normal-weight people worldwide. However, there is limited data on the GM profiles of obese and control Indian individuals. Fecal samples from 20 participants (10 obese, 10 control) underwent 16S rRNA gene sequencing. Anthropometric analysis confirmed significant differences in weight and BMI, with no significant variance in age or height. Taxonomic profiling and diversity indices (Chao1, Shannon, Simpson) were evaluated using the NCBI database. Sequencing identified 1,359 Operational Taxonomic Units. The obese cohort exhibited an elevated Firmicutes-to-Bacteroidetes (F/B) ratio, with Firmicutes increasing to 53.78% and Bacteroidetes decreasing to 25.54%. A significant reduction in Fusobacteria was observed in the Ob group (p = 0.034). Seven species were nominally enriched in obese subjects: Bifidobacterium catenulatum (p = 0.031), Anaerostipes hadrus (p = 0.014), Eggerthella lenta(p = 0.032), Bifidobacterium bifidum (p = 0.037), Clostridium butyricum(p = 0.012), Phascolarctobacterium sp.(p = 0.046), and Alistipes onderdonkii(p = 0.033). Rarefaction curves showed higher species richness in the control group, whereas PCA plots indicated greater community similarity (lower beta diversity) in the obese group. Obesity is associated with distinct microbial dysbiosis, characterized by a significant loss of Fusobacteria and an enrichment of SCFA-producing species. These specific taxonomic shifts, rather than broad diversity indices, provide a more sensitive signature for metabolic changes associated with obesity, supporting the energy harvest hypothesis.

Disruptions in the early-life gut microbiome have been reported to be associated with various health conditions. However, few studies have investigated the establishment of the gut microbiota in the immediate postnatal period. To characterize the gut microbiota of neonates within the first 100 hours after birth. A total of 512 healthy neonates born at a tertiary hospital were enrolled in this study. Stool samples were collected between March 2024 and December 2024 and categorized by delivery mode and stool collection time. Microbiota diversity and composition were assessed using full-length 16S rDNA sequencing. Stool samples from 140 neonates with predominant breastfeeding were analyzed. Of the 140 neonates, 70 (50%) were female, 66 (47.1%) were delivered by vaginal delivery (VD), 74 (52.9%) were delivered by cesarean section (CS), and 87 (62.1%) were firstborn. The time of stool collection ranged from 13.2 minutes to 109.82 hours. A significant difference in the time of stool collection was observed between CS and VD neonates (43.0 hours; 95%CI: 35.8-50.1 vs 32.4 hours; 95%CI: 27.8-36.9; P = 0.016). Gut microbiota analysis revealed that CS was associated with higher alpha diversity (Chao1 and Shannon indices; P < 0.001) and beta diversity (P < 0.001). A reduction in the gut microbiota diversity was observed 48 hours after birth (P < 0.001). Escherichia coli predominated in the VD samples. Bifidobacterium species, including Bifidobacterium longum, Bifidobacterium pseudocatenulatum, and Bifidobacterium bifidum, emerged significantly 48 hours after birth. CS neonates exhibited a higher relative abundance of skin- and environmental-associated taxa, including Staphylococcus, Streptococcus, and Pseudomonas, and markedly reduced colonization by Bifidobacterium species despite predominant breastfeeding. Despite predominant breastfeeding, CS neonates showed reduced colonization by Bifidobacterium and a greater abundance of skin- and environment-associated taxa within the first 100 hours of life. These findings indicate that delivery mode can be a key determinant of early microbial assembly and highlight the need for targeted strategies to support the establishment of beneficial microbiota in CS-delivered infants.

Postbiotic metabolites derived from probiotics bacteria have gained increasing attention as multitargeted anticancer agents with improved safety and stability compared to live microorganisms. In this study, we evaluated the anticancer activity of freeze-dried cell-free supernatants (CFS) obtained from Bifidobacterium bifidum and Lacticaseibacillus rhamnosus against human colorectal adenocarcinoma (Caco-2) cells and integrated functional, molecular, and omics-based analyses. Cytotoxicity assessment using the MTT assay demonstrated concentration-dependent growth inhibition, with IC₅₀ values of 94.38µg/mL and 395.19µg/mL for B. bifidum and L. rhamnosus CFS, respectively. Flow cytometry and morphological analyses confirmed apoptosis induction and G₀/G₁ cell-cycle arrest. Immunocytochemical evaluation revealed increased E-cadherin and caspase-3 expression accompanied by vimentin downregulation, indicating suppression of epithelial-mesenchymal transition (EMT). Genomic screening identified apoptosis-related proteases and metalloproteases, supported by three-dimensional structural prediction, suggesting a functional role in postbiotic bioactivity. Metabolomic profiling using HPLC and GC-MS revealed strain-specific signatures dominated by organic acids, volatile compounds, and fatty acids associated with oxidative, inflammatory, and apoptotic modulation. Collectively, these findings demonstrate that freeze-dried CFS exert anticancer effects through coordinated transcriptomic-independent, proteomic, and metabolomic mechanisms, highlighting their potential as multifunctional postbiotic candidates for colorectal cancer intervention.

Early life is a critical window for bone mineral deposition, immune system development, and neurodevelopment. This study evaluated the effects of four probiotic strains on bone growth and regulation of immune function and cognitive function using a growing mouse model, thereby providing preclinical evidence for potential applications in supporting infant and child health. Female BALB/c mouse pups received a daily oral gavage of 1 × 109 colony-forming units of four probiotic strains or saline from postnatal week 1 to week 5. At the experimental endpoint, bone microarchitecture was evaluated using micro-computed tomography and histomorphometric analysis. Lacticaseibacillus paracasei L578 increased femoral length and bone volume fraction, while Bifidobacterium bifidum B11 supplementation specifically improved cortical bone parameters. Mechanistic insights revealed that L. paracasei L578 upregulated serum concentrations of bone formation markers, namely osteocalcin, bone-specific alkaline phosphatase, and procollagen type I N-terminal propeptide, along with the transcription levels of key osteogenic transcription factors Bmp2, Runx2, and Osterix, which constitute a hierarchical cascade governing osteoblast lineage commitment and differentiation. Immunological assessments indicated that both L. paracasei L578 and B. animalis subsp. lactis BAL531 significantly increased colonic concentrations of immunoglobulin A, immunoglobulin G, and interferon-γ, suggesting their role in regulating the Th1/Th2 immune balance and promoting mucosal humoral immunity. In behavioral tests, L. paracasei L578 administration resulted in significantly longer time spent in the central area of the open field and a higher spontaneous alternation rate in the Y maze, indicating enhanced cognitive function. Collectively, these findings suggest that L. paracasei L578 enhances bone formation metabolism, stimulates longitudinal bone growth, increases bone mass, regulates mucosal humoral immunity, and improves cognitive function, positioning it as a promising probiotic for supporting development. © 2026 Society of Chemical Industry.

Given the global risks associated with antibiotic overuse, antibiotic-associated diarrhea (AAD) has become a widespread public health concern worldwide. Human milk oligosaccharides (HMO) and bifidobacteria are often used to prepare synbiotics to enhance their health effects. However, the synergistic effect of this combination in alleviating AAD remains unclear. In this study, lincomycin was used to establish a mouse model of AAD, and the effects of 3-fucosyllactose with 6'-sialyllactose, as well as Bifidobacterium bifidum, were evaluated. Further analyses examined the effects of HMO plus probiotics on the gut microbiota and its associated metabolites. In addition, the roles and underlying mechanisms of specific metabolites in AAD were investigated. The results indicate that HMO plus probiotics significantly ameliorate diarrheal symptoms in mice with AAD. This effect is reflected by increased body weight, markedly reduced diarrhea scores and fecal water content, significantly elevated aquaporin-3 (AQP3) expression, and substantially decreased levels of the inflammatory mediators tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β). Moreover, the combination of HMO and probiotics demonstrated superior efficacy compared with either intervention alone. Integrated microbiome-metabolomics analysis revealed that HMO plus probiotics significantly enriched Lactobacillus, a change that was potentially associated with increased levels of the gut metabolite indole-3-acetic acid (IAA). Indole-3-acetic acid plays a crucial role in intestinal barrier repair by promoting Muc2 expression and thereby enhancing the intestinal chemical barrier. Concurrently, IAA contributes to the restoration of the mechanical barrier by regulating claudin-1 protein expression. Furthermore, the effects of IAA were significantly attenuated following intervention with aryl hydrocarbon receptor (AhR) inhibitors, indicating that IAA exerts its actions via activation of AhR. In summary, this study suggests that HMO plus probiotics may alleviate AAD by promoting IAA production via the gut microbiota, thereby activating the IAA/AhR/Muc2/claudin-1 axis. This research provides novel insights into microbiota-based interventions targeting intestinal health.

A detoxifying multi-strain probiotic formula attenuates toxicity induced by heavy metals and phthalates.

The immune system is a highly coordinated network that defends the host against pathogens and maintains physiological homeostasis. Probiotics have emerged as promising immunoregulatory agents; however, the mechanisms by which they modulate innate and adaptive immune responses remain incompletely understood. In this study, we investigated the immunomodulatory potential of Bifidobacterium bifidum BGN4 using in vitro assays and a cyclophosphamide (CP)-induced immunosuppressed mouse model. In vitro, B. bifidum BGN4 treatment markedly enhanced natural killer (NK) cell cytotoxicity against YAC-1 lymphoma target cells and activated macrophages, as evidenced by elevated nitric oxide production, increased secretion of pro-inflammatory cytokines (IL-1β, IL-6, and TNF-α), and upregulated expression of phagocytosis-related genes (Marco, Msr1, and Cd14). These effects were accompanied by activation of MAPK and NF-κB signaling pathways in macrophages. In vivo, oral administration of B. bifidum BGN4 significantly mitigated CP-induced reductions in body weight, spleen and thymus indices, and leukocyte counts. Moreover, B. bifidum BGN4 enhanced splenocyte proliferation under both basal and mitogen-stimulated conditions and improved NK cell cytotoxic activity in immunosuppressed mice. Collectively, these findings demonstrate that B. bifidum BGN4 promotes both innate and adaptive immune activation, effectively counteracting CP-induced immunosuppression. This probiotic strain represents a promising functional food ingredient for immune enhancement and recovery.

Gut microbiota and its metabolites are revealed to affect metabolic dysfunction-associated fatty liver disease (MAFLD) progression. Precedingly, we proved bifidobacterium bifidum BGN4 fractions declined sterol regulatory element-binding transcription factor 1 (SREBP1) to suppress ferroptosis during MAFLD. This study explored whether agmatine, a key metabolite induced by BGN4, affected MAFLD development. High-fat diet (HFD)-fed mice or palmitate (PA)-stimulated MIHA hepatocytes were treated with agmatine. General conditions of the mice were evaluated by behavioral tests, body weight, liver weight, and plasma biochemical parameters, while hepatic steatosis was observed with H&E staining. Hepatocyte cytotoxicity was assessed by lactate dehydrogenase assay, and mitochondrial dysfunction was evaluated with ATP content and oxygen consumption rate. Ferroptosis was estimated by lipid peroxidation, iron, and expressions of related proteins. Immunohistochemistry, immunofluorescence, western blot, and RT-qPCR were used for detecting protein and mRNA levels. Molecular mechanism was investigated using RNA immunoprecipitation, methylated RNA immunoprecipitation, luciferase reporter assay, and RNA stability assay. In the HFD-induced MAFLD mouse model, agmatine treatment alleviated metabolic abnormalities, depression-related behavior, hepatic damage, and ferroptosis, and decreased Iba1 and GFAP in cortex. In the in vitro model of PA-induced hepatocytes, agmatine mitigates cytotoxicity, mitochondrial dysfunction, and ferroptosis. Increases of SREBP1, RNA binding motif protein 15 (RBM15), and IGF2 mRNA binding protein2 (IGF2BP2) in MAFLD models were rescued by agmatine. RBM15/IGF2BP2 knockdown inhibited m6A modification and stability of SREBP1 mRNA. SREBP1 overexpression neutralized the protective role of agmatine, which was overturned by silencing RBM15/IGF2BP2. Agmatine relieved MAFLD via inhibiting RBM15/IGF2BP2-mediated SREBP1 m6A modification.

Obesity, characterized by excess body fat accumulation, is closely linked to the alteration of gut microbiota, which contribute to systemic inflammation. Probiotics intervention has emerged as a promising strategy favorably modulating gut microbiota composition in obese individuals accompanied by improvements in metabolic parameters. The objective of this study was to evaluate whether supplementation with Bifidobacterium bifidum BGN4 alters gut microbiota composition and to assess its associated effects on circulating zonulin, a marker of intestinal permeability, as well as metabolic parameters in individuals with excess adiposity. This randomized, double-blind, placebo-controlled trial involved 60 adults with excess body fat (body fat percentage ≥ 20% for males and ≥ 28% for females). A total of 60 adults were enrolled, and sex- and age-stratified randomization allocated 30 participants to each group. Participants received one capsule of B. bifidum BGN4 (9 × 109 colony forming unit) or matched placebo for 8weeks. Blood samples were analyzed for zonulin, TNFα, hs-CRP, glucose, insulin, lipid profiles, and total antioxidant capacity. Fecal samples were analyzed to determine alterations in gut microbiota composition. A total of 58 participants, with 29 individuals in each group, successfully completed the 8-week intervention. Supplementation with B. bifidum BGN4 did not result in significant changes in BMI, body fat percentage, or the primary outcome, hs-CRP. In contrast, significant improvements were observed in serum zonulin (between-group differences: -1.61 ± 2.69ng/mL), TNFα (between-group differences: -0.17 ± 0.26pg/mL), and fasting insulin (between-group differences: -3.52 ± 10.25 μIU/mL). The probiotic intervention modulated the enrichment of several taxa, including Bacteroides coprocola, Bifidobacterium catenulatum group, Lactiplantibacillus plantarum group, and Prevotella stercorea. In addition, several microbial taxa demonstrated correlations with metabolic and inflammatory parameters. No adverse effects were observed, as indicated by stable liver enzyme concentrations, blood pressure, and gastrointestinal symptoms. The results indicate that B. bifidum BGN4 may serve as a preventive strategy for metabolic disorders in individuals with excess adiposity through the maintenance of gut microbial balance and intestinal barrier integrity thereby potentially mitigating inflammation and metabolic stress. Clinical trial registration number: KCT0010817. Date of registration: July 28, 2025.

Disialyllacto-N-tetraose (DSLNT), ahuman milkoligosaccharide implicated in neonatal immunity and necrotizing enterocolitisprevention, contains both Neu5Acα(2→3)­Gal and Neu5Acα(2→6)­GlcNAcmotifs. Its synthesis is hindered by challenging α-sialylation.Here we report the total chemical synthesis of DSLNT via simultaneousdouble sialylation using macrobicyclic donors, achieving completestereocontrol. A convergent [4 + 2] glycosylation enabled efficientassembly. The route furnished ∼50 mg of free-reducing DSLNT,enabling enzymatic studies that revealed preferential cleavage ofα­(2→3)-linkage by Bifidobacterium bifidum sialidase SiaBb2.

Advocating gut-retina connection and microbiota mediated pathways in management of age-related macular degeneration: Preclinical to clinical perspective.

Background: Secretory immunoglobulin A (sIgA) is central to epithelial defense and host-microbe homeostasis. Probiotic products positioned for immune support frequently underperform when strain selection, storage resilience, and intestinal delivery are not optimized. Methods: We reorganized a formulation-development and preclinical dataset describing the BALIMONT immune-support platform and integrated representative published evidence on IgA-oriented probiotic, postbiotic, and prebiotic interventions. The BALIMONT system combined Bifidobacterium bifidum ATCC 29521, Bifidobacterium longum DSM 20219, and Lacticaseibacillus rhamnosus ATCC 7469 with a heat-inactivated Lactococcus lactis-derived postbiotic fraction, a prebiotic support matrix, and staged protective encapsulation. Results: Within the development dataset, the 2:2:2 tri-strain ratio yielded the strongest in vitro response, with secretory IgA of 21.35 μg/mL and probiotic proliferation of 5.62-fold. A 1:1 postbiotic-to-probiotic cell-count ratio maximized both IgA induction and 30-day viable-count retention. In mice, the 200 mg/kg BALIMONT group exceeded a same-category comparator by 39.68% for serum total IgA, 56.52% for intestinal mucosal sIgA, and 53.01% for fecal sIgA. Viable-count retention remained 86.72% after 6 months at 25°C/60% RH. Published clinical and translational evidence further supports the biological plausibility of IgA-associated mucosal benefits from probiotic and synbiotic strategies. Conclusions: The BALIMONT platform can be interpreted as a rationally balanced probiotic-postbiotic system in which strain synergy, postbiotic complementation, prebiotic support, and staged delivery converge on IgA-associated mucosal immunity. These findings support continued translational development and justify subsequent human validation.

Anxiety is a prevalent non-motor symptom of Parkinson’s disease (PD), yet treatment options remain limited. This randomized, double-blind, placebo-controlled trial evaluated the effects of a 12-week probiotic supplement containing nine bacterial strains (Bifidobacterium bifidum W23, Bifidobacterium lactis W51 and W52, Lactobacillus acidophilus W37, Levilactobacillus brevis W63, Lacticaseibacillus casei W56, Ligilactobacillus salivarius W24, and Lactococcus lactis W19 and W58) on anxiety symptoms in 61 individuals with PD and clinically significant anxiety. Both the probiotic (n = 30) and placebo (n = 31) groups showed significant within-group improvements on the Parkinson Anxiety Scale, with no significant between-group differences. The probiotic group showed a statistically significant improvement on the Montreal Cognitive Assessment compared with placebo (adjusted mean difference of 1.1 points; 95% confidence interval: 0.04–2.1; p = 0.043). No treatment effects were seen on other secondary outcomes, including depression, constipation, or motor symptoms. No significant between-group differences in gut microbiota composition or systemic inflammatory markers were observed. While the multi-strain probiotic did not reduce anxiety more than placebo, its potential cognitive effects warrant further investigation in larger trials. This trial was registered on ClinicalTrials.gov (NCT03968133) on May 28, 2019.

In scientific literature, it is relatively rare to find information on healthcare professionals' current knowledge of probiotics, prebiotics, and gut microbiota. The aim of our study was to assess healthcare professionals' knowledge of gut microbiota, probiotics, and prebiotics in the city of Kénitra, Morocco. The data was collected via an online questionnaire, which we distributed through social media. A total of 143 healthcare professionals (78.3% women and 21.7% men) responded to this questionnaire. The questionnaire concerned knowledge of probiotics and prebiotics. Most respondents rated their knowledge of probiotics, prebiotics, and gut microbiota as average (40%) or poor (39%), while others rated their knowledge as good (11%) and only 2% had very good knowledge, with the remainder (8%) having no knowledge. The correct definition of probiotics chose 67.1% of respondents, broken down as follows: 80.4% of general practitioners, 76.9% of specialists, and 57% of nurses. Lactobacillus acidophilus (65%) and Bifidobacterium bifidum (50.3%) are the two species best known to respondents as probiotic strains. Furthermore, the most popular prebiotic is fructooligosaccharide (51%), followed by galactooligosaccharide (42.7%) then inulin (36.4%) and finally betaglucan (14%). Among professionals 60.1% prescribed probiotics and/or prebiotics for diarrhea, followed by antibiotics (47.6%) then constipation (39.2%) and 21% of respondents recommended them for diabetes, 18.9% for obesity while only 3.5% used them for other pathologies. This online survey revealed the current knowledge of healthcare professionals regarding probiotics, prebiotics and gut microbiota and highlights the importance of educating and training them through targeted learning programs.

To evaluate the effects of probiotics Bifidobacterium bifidum G9-1 (BBG9-1) on gastrointestinal symptoms in patients with Type 2 diabetes mellitus (T2DM) with diarrhoea or constipation. In a 12-week open-label randomised controlled trial, 100 participants were randomized 1:1 to a control or BBG9-1 group, with 12 mg of Bifidobacterium administered daily. The primary endpoint was the change in total Gastrointestinal Symptom Rating Scale (GSRS) score (range: 1-7) in the full analysis set (BBG9-1: n = 43; control: n = 51). BBG9-1 intake significantly improved the GSRS total score (from 2.22 ± 0.67 to 1.83 ± 0.62) compared with control (from 2.08 ± 0.67 to 2.06 ± 0.63; between-group difference, -0.34 [95% CI: -0.55 to -0.14; p = 0.001]). Subgroup analyses indicated greater GSRS total score improvement in women, participants with constipation, aged ≥ 65 years, and BMI < 25 kg/m2. Among the GSRS subscales, BBG9-1 showed a significantly greater mean reduction in constipation scores than control (-0.79 ± 1.38 vs. -0.13 ± 1.04; p = 0.013). Although the mean change in diarrhoea scores did not differ significantly (-0.42 ± 1.14 vs. -0.06 ± 1.08; p = 0.14), the follow-up diarrhoea score was significantly lower with BBG9-1 (2.04 ± 1.00 vs. 2.58 ± 1.44; p = 0.049). Through gut microbiota analysis, the relative abundance of genera Phocaeicola increased significantly in the BBG9-1 group (from 15.98 ± 13.75 to 19.56% ± 14.79% vs. from 18.14 ± 14.17 to 18.21 ± 13.58, p = 0.042). Assessment of faecal short-chain fatty acids (SCFAs) revealed no significant changes associated with BBG9-1 administration. The incidence of adverse effects were similar between the groups. BBG9-1 administration significantly improved gastrointestinal symptoms in patients with T2DM. Japan Registry of Clinical Trials (jRCTs051220127).

While synbiotic interventions show promise in alleviating food allergy (FA), targeted optimization of probiotic-prebiotic combinations for enhanced immunomodulatory metabolite production remains insufficiently explored. This study constructed a novel synbiotic containing five human milk oligosaccharides (5-HMOs), Bifidobacterium bifidum R0071, bovine whey protein hydrolysate, and tryptophan, which demonstrated efficient production of indole-3-lactic acid (ILA) in vitro. In an ovalbumin (OVA)-induced FA mouse model, the synbiotic significantly alleviated allergic symptoms, reduced OVA-sIgE levels, restored Th2/Th17 immune balance, improved intestinal barrier integrity, and activated the aryl hydrocarbon receptor. 16S rRNA sequencing revealed that this synbiotic treatment enriched beneficial gut bacteria, including Bifidobacterium, Akkermansia, and Lactobacillus johnsonii. Targeted metabolomics confirmed elevated intestinal ILA levels, which were closely correlated with the alleviation of FA. Collectively, these findings demonstrate that the 5-HMOs synbiotic alleviates FA by modulating the gut microbiota and enhancing ILA production, offering novel insights into the targeted development of synbiotic-based interventions against FA.

Background/Objectives: Atopic dermatitis (AD) is associated with gut dysbiosis linked to early-life antibiotic use and Staphylococcus aureus colonization. Gum Arabic (GA), a prebiotic, may modulate this dysbiosis and influence AD-related microbial balance. This study evaluated whether GA could support AD-associated probiotics-Lactobacillus casei, Bifidobacterium bifidum, and Bifidobacterium infantis-under amoxicillin- or azithromycin-containing conditions, examined the response of S. aureus under the same screening conditions, and developed GA-phospholipid-based semisolid carriers for topical application. Methods: Probiotic strains were cultured with 1-5% GA in the presence and absence of antibiotics, and viable cell counts were assessed. Sixteen topical formulations containing propylene glycol or isopropyl myristate in a hydrogenated phosphatidylcholine base were prepared and screened for rheological properties and galactose release using in vitro release testing (IVRT) and HPLC-UV. Results: GA at 1-2% concentrations promoted probiotic growth in antibiotic-free conditions. GA preserved B. infantis viability under azithromycin exposure in this in vitro screening model. For S. aureus, numerical CFU differences were observed between antibiotic-only and GA-containing conditions; however, the present screening design was not intended to determine antibiotic interaction outcomes. Formulations F14 (2% GA + 7% IPM) and F15 (3% GA + 7% IPM) exhibited optimal spreadability. IVRT showed that 6 h cumulative galactose release varied by formulation (F6 > F10 > F14 > F15). Conclusions: GA demonstrated dose-dependent prebiotic activity and preserved B. infantis viability under azithromycin exposure in this in vitro screening model. For S. aureus, the observed CFU differences between antibiotic-only and GA-containing conditions should be considered exploratory only and do not allow for conclusions regarding interference with antibiotic efficacy. Optimized GA-HPC systems with suitable rheological and release characteristics represent promising candidates for further preclinical investigation.

The capacity of Lactobacillus casei, Lactobacillus rhamnosus, Bifidobacterium bifidum, and Bacillus subtilis to increase the content of gymnemagenin in Gymnema spp. R.Br was investigated on two samples of G. sylvestre and two samples of G. latifolium by incubating these samples for 8 days with each strain of bacteria. The difference in content of gymnemagenin before and after incubation was determined by using a validated HPLC method meeting all requirements of AOAC International on analytical method performance. The four strains of bacteria were found to increase the content of gymnemagenin in the tested samples after incubation, but the change caused by B.subtilis was not statistically significant. No significant strain-to-strain difference in the capacity to improve the content of gymnemagenin was found among the four strains of bacteria.

Adenine and myristic acid from Limosilactobacillus reuteri enhance growth and freeze-drying tolerance of Bifidobacterium bifidum CCFM1167