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Probiotic extracellular vesicles are biochemically active structures responsible for biological effects elicited by probiotic bacteria. Lactobacillus spp., which are abundant in the human body (e.g., gut), are known to have anti-inflammatory and antimicrobial properties, and are commonly used in food products, supplements, and in discovery research. There is increasing evidence that Lactobacillus–derived extracellular vesicles (LREVs) have potent immunomodulatory capacity that is superior to probiotics themselves. However, key mechanistic insights into the process that controls production and thus, the function of LREVs, are lacking. Currently, it is unknown how the probiotic culture microenvironment orchestrates the type, yield and function of LREVs. Here, we investigated how multifactor modulation of the biomanufacturing process controls the yield and biological functionality of the LREVs. To achieve this, we selected Lacticaseibacillus rhamnosus as the candidate probiotic, initially cultivated under traditional culture conditions, i.e., 100% broth concentration and pH 5.5. Subsequently, we systematically modified the culture conditions of the probiotic by adjusting three critical process parameters: (1) culture medium pH (pH 3.5, 5.5 and 7.5), (2) growth time (48 and 72 h), and (3) broth concentration (50% and 10% of original broth concentration). EVs were then isolated separately from each condition. The critical quality attributes (CQA) of LREVs, including physical characteristics (size, distribution, concentration) and biological composition (protein, carbohydrate, lipid), were analysed. Functional impacts of LREVs on human epidermal keratinocytes and Staphylococcus aureus were also assessed as CQA. Our findings show that the production of LREVs is influenced by environmental stresses induced by the culture conditions. Factors like broth concentration, pH levels, and growth time significantly impact stress levels in L. rhamnosus, affecting both the production and composition of LREVs. Additionally, we have observed that LREVs are non-toxicity for keratinocytes, the major cell type of the epidermis, and possess antimicrobial properties against S. aureus, a common human skin pathogen. These properties are prerequisites for the potential application of EVs to treat skin conditions, including infected wounds. However, the functionality of LREVs depends on the culture conditions and stress levels experienced by L. rhamnosus during production. Understanding this relationship between the culture microenvironment, probiotic stress response, and LREV characteristics, can lead to the biomanufacturing of customised probiotic-derived EVs for various medical and industrial applications.

The objective of the study was to systematically assess whether the oral probiotic Lactobacillus rhamnosus (LGG) improves symptoms of irritable bowel syndrome in children. Four randomised clinical trials met the inclusion criteria, a total of 252 patients. Success of treatment was reported in 50 out of 82 patients (58.82%) from the LGG group, compared with 28 from 82 (34.14%) in the placebo group (p = 0.05). The number of pain episodes was lower in the LGG group compared with the placebo group (n = 117, 95% CI: 1.50 [–2.03; –0.97], p < 0.05). The use of LGG resulted in a decrease in the perception of pain intensity in the study population overall (n = 219, 95% CI: –0.61 [–1.13; –0.09], p < 0.05). A higher dosage of LGG was less effective than a lower dosage in improving the overall treatment success (p = 0.05). The dosage of LGG was not statistically significant in improving pain severity (p = 0.12).

The development of baby spinach as a vehicle to transfer Lactobacillus plantarum 299v (LP299v) and Lactobacillus rhamnosus GG (LGG) is quite promising and may address the research regarding the absence of suitable whole vegetable carriers in the current probiotic food industry. The objective of this study was to observe the effects of food storage and preparation on Lp299v and LGG viability in baby spinach before consumption. The strains were sequentially introduced into baby spinach by dipping leaves in probiotic suspension to achieve an attachment of approximately 8 log 10 CFU/g spinach. Then, probiotic viability was tested using serial dilutions. Furthermore, data processing and ANOVA during 7-day storage, with or without salad dressing, were performed using Tukey’s test. In the 7-day storage trials, LP299v and LGG viability on baby spinach declined after 7 days with significant differences by 0.19 and 0.39 log10 CFU/g, respectively. In salad dressing trials, LP299v (p value = 0.79 > 0.05) and LGG (p value = 0.58 > 0.05) survivability on baby spinach after the addition of salad dressing fluctuated approximately 8.27 and 8.40 log10 CFU/g with no statistically significant difference, respectively. LP299V and LGG viability on baby spinach in both trials was greater than 8 log10 CFU/g and close to FDA requirements, showing that food storage and preparation do not affect their viability and can be used commercially.

Non-alcoholic fatty liver disease (NAFLD) is a significant consequence of metabolic dysfunction, often associated with changes in the intestinal microbiota. Prebiotics and probiotics have shown promise in NAFLD management. This study evaluated a silymarin-based herbal remedy with piperine and fulvic acid, alongside a probiotic blend of Bifidobacterium adolescentis, Bifidobacterium bifidum, Lactobacillus casei, and Lactobacillus rhamnosus. Using a NAFLD mouse model induced by a high-fat and high-fructose diet, we assessed biochemical parameters, liver function, glucose levels, and conducted histological analysis. Stool samples underwent 16S rRNA metagenomic analysis to explore changes in microbiota composition. Mice on the high-fat diet exhibited elevated lipids, liver enzymes, and glucose, with reduced high-density lipoprotein levels (with p value

Recently, non-dairy based probiotic beverages has a progressive market, in this study the effect of ultrasound on the fermentation kinetics and quality of millet-based probiotic beverages (MPB) was carried out. White finger millet (Eleusine coracana L.) variety KMR-340 was used in this study. Two treatment conditions including ultrasound (US) treatment before inoculation (0.53–1.59 W/mm2 for 5–15 min) and US treatment after inoculation (0.21–0.35 W/mm2 for 1–5 min) were taken as a pretreatment. The optimum condition for US treatment before inoculation was identified as 0.99 W/mm2 intensity and 11.09 min, in the case of US treatment after inoculation, it was identified as 0.29 W/mm2 intensity and 2.63 min treatment time of US treatment. In the US treatment after inoculation, requires the lowest fermentation time which is 2.6-fold times lower than the untreated samples. Structural changes in millet protein and other molecules by the US alter the techno-functional behavior of MPB like water holding capacity (WHC) and viscosity. The Weibull model explains the fermentation kinetics of MPB, the rate constant of lactic acid was higher in US treatment after inoculation explaining the probiotic microbial proliferation by low-intensity US treatment. During fermentation, change in lactic acid follows three stages: rapid change, constant period, and gradual change. These findings prove that changes in US treatment highly affect the fermentation kinetics of probiotic microbes, and the US treatment can be used to alter the physicochemical changes in probiotic beverages.

This study investigated the effects of Lactobacillus rhamnosus fermentation products added to feed on common carp (Cyprinus carpio) on growth, gut and liver health, and gut microbiota. Common carp with an average weight of 1.95±0.4 g were fed five experimental diets: normal diet (normal) and normal diet supplemented with Lactobacillus rhamnosus fermentation products (abbreviation: 0.2% GCC-3, 0.3% GCC-3, 0.4% GCC-3 and 0.5% GCC-3, respectively) for 8 weeks. The results showed that the GCC-3 fermentation product did not affect the growth phenotype of common carp. The diet supplemented with 0.3% GCC-3 significantly reduced the serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels of common carp compared to the control group. Diets with GCC-3 fermentation products significantly reduced liver triacylglycerol (TAG) levels. Diets supplemented with 0.3% GCC-3 fermentation products significantly downregulated the expression of liver liposynthesis gene (fas) and significantly upregulated the expression of liver lipolysis gene (cpt1). Diets supplemented with GCC-3 significantly reduced the serum lipopolysaccharide (LPS) levels of common carp compared to the control group. In addition, dietary supplementation with 0.3% GCC-3, 0.4% GCC-3, or 0.5% GCC-3 all increased the abundance of Cetobacterium and decreased the abundance of Acinetobacter. In summary, dietary supplementation with 0.3% GCC-3, 0.4% GCC-3, and 0.5% GCC-3 reduced liver lipid deposition, and improved gut microbiota homeostasis of common carp. The best positive effect was observed in common carp fed a 0.3% GCC-3 supplemental diet.

To date, probiotic medicinal products have a significant number of clinical studies, which is the basis for the formation of an evidence base and their inclusion in modern treatment standards. However, ideas about the optimal composition of a probiotic preparation and effective dosage remain problematic issues in modern medicine. Choosing an effective probiotic in gynecological practice is a particularly difficult and important issue. Objective of the review: to analyze the available clinical studies aimed at identifying strains of beneficial bacteria, their dosage, possible combinations in order to choose the optimal composition of a probiotic agent for the treatment of dysbiosis, including vaginal. Materials and methods. Modern methods recommended for theoretical analytical research were used: method of analysis, method of synthesis, method of induction. An electronic search was conducted in bibliographic databases, including PubMed, CENTRAL, MEDLINE, Embase, and CINAHL. 58 scientific publications were analyzed. Inclusion criteria were the following keywords: probiotics, optimal dosage, optimal composition, dysbiosis. Results. The most important probiotic microorganisms for restoring health in women are Lactobacillus acidophilus, L. crispatus, L. jensenii, L. gasseri, L. rhamnosus. Each of these lactobacilli maintains the normal vaginal pH, produces hydrogen peroxide, suppresses growth and prevents the reproduction of obligate anaerobes and opportunistic microorganisms when interacting with the peroxidase of cervical mucus. The mechanism of action of the above-mentioned probiotic microorganisms was analyzed. It was established that the effective dose of each of the components should not be less than 1-2 billion colony forming units. Conclusions. The analysis of scientific publications in the bibliographic databases was sufficient to establish that the oral use of probiotics in the treatment of gynecological patients is effective. The optimal components of the probiotic agent, which are effective for the therapy of vaginal dysbiosis, are L. crispatus, L. gasseri, L. jensenii, L. rhamnosus, L. acidophilus.

(1) Background: Given the autoimmune nature of Alopecia Areata (AA) and the immunomodulatory properties of probiotics, this trial was conducted to evaluate the efficacy of a probiotic mixture, consisting of Lactobacillus rhamnosus and Bifidobacterium longum strains, as an adjuvant treatment in a group of AA patients. (2) Method: This study was a 24-week, randomized, double-blind, placebo-controlled clinical trial. Twenty-six patients with AA were included in this study, and their clinical progression, along with changes in gut and skin microbiota, were analyzed. (3) Results: A higher proportion of AA patients treated with the probiotic formula showed improvement compared to the placebo group, based on both the reduction in the number of AA plaques (56% vs. 30%) and the affected scalp surface area (45% vs. 20%). For “activity”, “inactivity”, and “regrowth”, an improvement in 55%, 67%, and 55% of patients was, respectively, observed in the probiotic group, compared to 50%, 40%, and 30% in the placebo group. No changes were observed in the gut microbiota during the intervention period. Regarding skin microbiota, changes were detected in the probiotic group, with reductions in characteristic genera during the study. (4) Conclusions: To our knowledge, this is the first clinical trial assessing the efficacy of a probiotic product in patients with AA. This probiotic mixture in a routine clinical practice setting appears to improve the course of patients. In addition, the skin microbiota of scalp lesions was modified using the probiotic treatment.

Alcoholic liver damage is caused by long-term or heavy drinking, and it may further progress into alcoholic liver diseases (ALD). Probiotic supplements have been suggested for the prevention or improvement of liver damage. This study was designed to consider the ameliorative effects of Lactobacillus rhamnosus NKU FL1-8 isolated from infant feces against alcoholic liver damage. The mice were gavaged with a 50% ethanol solution and treated with 109 CFU of L. rhamnosus NKU FL1-8 suspension. The factors for liver function, oxidative stress, inflammation, gut microbiota composition, and intestinal barrier integrity were measured. The results showed that L. rhamnosus NKU FL1-8 could decrease the levels of aspartate aminotransferase (AST) to 61% and alanine aminotransferase (ALT) to 50% compared with ethanol given by gavage. It could inhibit the expression level of malondialdehyde (MDA), increase superoxide dismutase (SOD), glutathione (GSH) to relieve oxidative stress, and down-regulate the cytokines to decrease hepatic inflammation. After treatment, the level of triglycerides was reduced, and the expression levels of adenosine 5′-monophosphate (AMP)-activated protein kinase (AMPK) and the peroxisome proliferators-activated receptor-α (PPAR-α) pathway were up-regulated. Additionally, the 16S rRNA sequencing analysis showed that L. rhamnosus NKU FL1-8 increased the relative abundance of Lactobacillus, Ruminococcaceae, etc. At the same time, L. rhamnosus NKU FL1-8 could significantly reduce lipopolysaccharides (LPS) and enhance intestinal tight junction proteins. These results demonstrated that L. rhamnosus NKU FL1-8 could reduce the level of oxidative stress, fat accumulation, and liver inflammation caused by alcohol in the host. The underlying mechanism could be that L. rhamnosus NKU FL1-8 inhibits LPS by regulating the gut microbiota and repairing the intestinal barrier. Thereby, these findings support L. rhamnosus NKU FL1-8 as a potential functional food for the relief of ALD.

A total of 200 samples of chicken products collected randomly from Quesna and El-Bagour in Menoufia governorate. The collected samples were represented by chicken breast, thigh, gizzard and liver (50 of each) and subjected to bacteriological examination for detection of Helicobacter species particularly, Helicobacter pylori. Further, the effect of L. rhamnosus culture to control such serious pathogen contaminating chicken products was studied. Helicobacter spp detected in 4%, 6%, 12% and 20% of breast, thigh, gizzard and liver, respectively and H. pylori represent 4%, 4% 6% and 10% from this examined samples, respectively. H. pylori inoculated chicken fillet (1×106 /g) which are not treated with  Lactobacillus rhamnosus (control +ve) showed increase in prevalence of H. pylori 1.0×106, 1.3×106, 1.9×106 and 2.4×106 at zero time, 1st day, 2nd day and 3rd day, respectively.  While chicken fillets treated with L. rhamnosus (1x106 /g) Effect on prevalence of H. pylori inoculated into chicken fillets with a reduction percent of 35%, 52% and 69% but  L. rhamnosus (1x107 /g) showed remarkable reduction of H. pylori prevalence with a reduction percent of 54%, 67% and 81% at 1st day, 2nd day and 3rd day, respectively. L. rhamnosus effected on virulence genes of H. pylori isolated from treated chicken fillet as L. rhamnosus (1x106/g) destructed cagA gene and L. rhamnosus (1x107/g) destructed cagA and vacA genes so using of Lactobacillus rhamnosus culture as a method of control has a good effect on prevalence of H. pylori and its virulence genes in chicken fillets and L. rhamnosus (1x107 /g) more effective than L. rhamnosus (1x106 /g).Â

A total of 200 samples of chicken products collected randomly from Quesna and El-Bagour in Menoufia governorate. The collected samples were represented by chicken breast, thigh, gizzard and liver (50 of each) and subjected to bacteriological examination for detection of Helicobacter species particularly, Helicobacter pylori. Further, the effect of L. rhamnosus culture to control such serious pathogen contaminating chicken products was studied. Helicobacter spp detected in 4%, 6%, 12% and 20% of breast, thigh, gizzard and liver, respectively and H. pylori represent 4%, 4% 6% and 10% from this examined samples, respectively. H. pylori inoculated chicken fillet (1×106 /g) which are not treated with  Lactobacillus rhamnosus (control +ve) showed increase in prevalence of H. pylori 1.0×106, 1.3×106, 1.9×106 and 2.4×106 at zero time, 1st day, 2nd day and 3rd day, respectively.  While chicken fillets treated with L. rhamnosus (1x106 /g) Effect on prevalence of H. pylori inoculated into chicken fillets with a reduction percent of 35%, 52% and 69% but  L. rhamnosus (1x107 /g) showed remarkable reduction of H. pylori prevalence with a reduction percent of 54%, 67% and 81% at 1st day, 2nd day and 3rd day, respectively. L. rhamnosus effected on virulence genes of H. pylori isolated from treated chicken fillet as L. rhamnosus (1x106/g) destructed cagA gene and L. rhamnosus (1x107/g) destructed cagA and vacA genes so using of Lactobacillus rhamnosus culture as a method of control has a good effect on prevalence of H. pylori and its virulence genes in chicken fillets and L. rhamnosus (1x107 /g) more effective than L. rhamnosus (1x106 /g).Â

Background and Objectives: Colorectal cancer (CRC) is a prevalent form of cancer worldwide. Recent studies suggest that postbiotics derived from probiotic bacteria have the potential as an adjunct therapy for CRC. This study investigates the anti-cancer effects of Bifidobacterium breve (B. breve) and Lactobacillus rhamnosus (L. rhamnosus) postbiotics on the HT-29 cell line. Materials and Methods: Through MTT and scratch assay, we investigated the anti-proliferation and anti-migration effects of B. breve and L. rhamnosus postbiotics on HT-29 cells. Furthermore, postbiotic-mediated apoptosis was assessed by analyzing the expression of Bax, Bcl-2, and caspase-3. We also investigated the effects of B. breve postbiotics on the expression of three important genes involved in metastasis, including RSPO2, NGF, and MMP7. Consequently, we validated the expression of selected genes in twelve adenocarcinoma tissues. Results: The results demonstrated the significant impact of postbiotics on HT-29 cells, highlighting their ability to induce anti-proliferation, anti-migration, and apoptosis-related effects. Notably, these effects were more pronounced using B. breve postbiotics than L. rhamnosus. Additionally, B. breve postbiotics could inhibit metastasis through upregulation of RSPO2 while downregulating NGF and MMP7 expression in HT-29 cells. Conclusion: Our research suggests that postbiotic metabolites may be effective biological products for the prevention and treatment of cancer.

β-lactoglobulin (β-Lg) in milk is the main allergen that causes intestinal inflammation. Our previous study reported that Lactobacillus rhamnosus LZ260E (LZ260E) showed potential anti-allergic activity. However, the specific mechanism by which LZ260E exerts its anti-allergic activity remains unclear. This study investigated the effect of LZ260E on β-Lg-induced milk allergen mice model. The results showed that oral administration of LZ260E reduced spleen and thymus indices in mice. In addition, oral administration of LZ260E decreased the serum levels of IgE, histamine, IL-4, and monocyte chemotactic protein-1 (MCP-1), upregulated the expression of IFN-γ, IL-10, TNF-α, and TGF-β, and repaired colon damage. It was also found that LZ260E regulated alterations in the intestinal microbiota caused by allergies. These results showed that LZ260E has anti-allergic properties by regulating the gut microbiota and Th1/Th2 immunological balance, implying that it may be a functional probiotic against cow's milk caused allergies.

Objective: Phytochemistry, microbiology, flavor and functional properties of fermented papaya juice were examined in relation to Lactobacillus plantarum (Lp), Lactobacillus rhamnosus (Lr), Lactobacillus acidophilus (La) and their mixed strains (Lp+Lr、Lp+La、Lr+La), in order to develop a probiotic papaya beverage with improved functionality and health-promoting properties. Methods: In thisstudy, changes in pH, total soluble solids, titratable acid, viable cells, glucose, fructose, sucrose, organic acids and volatile compounds were determined. The contents of total phenolics and total flavonoids, as well as the inhibitory activity against α-glucosidase and tyrosinase enzymes during the fermentation process, were also investigated. Results: The single-and mixed strain cultures showed similar changes during the 72 h fermentation period, exhibiting significantly decreased pH and increased acidity. The contents of lactic acid and succinic acid greatly increased, whereas that of citric acid and malic acid decreased significantly. After fermentation, the content of total phenolics and flavonoids increased in papaya juice. Pearson's correlation analysis revealed that the metabolism of phenolics likely contributed to enhancing the inhibitory activity against α-glucosidase and tyrosinase enzymes. Among all fermentation groups, the papaya juice fermented by Lp exhibited the highest contents of lactic acid, total phenolics, and flavonoids, showing the strongest inhibitory effect against α-glucosidase and tyrosinase enzyme. Additionally, six types of aromatic compounds were primarily detected in unfermented papaya juice, with butyric acid content being the highest (89.742%), followed by linalool. After fermentation, a total of 25 aromatic compounds were identified in the six fermented juice samples, and the content of butyric acid declined significantly, which reduced the unpleasant odor of papaya. In all fermentation groups, the Lp single-strain culture generated the most volatile components (18 types), while the Lp+Lr mixed culture achieved the lowest (5 types). Although the Lp+Lr mixed-strain culture had the highest number of viable bacteria [7.61 lg (CFU/mL)], the Lp single-strain culture may be preferable overall. Conclusion: The Lp single culture could improve the sensory characteristics of papaya juice and enhance its beneficial properties.

BackgroundThe depletion of beneficial bacteria in the gut has been found in patients with acne vulgaris, and in previous studies, the supplement of Lactobacillus rhamnosus led to the improvement of adult acne. Nevertheless, the potential mechanism of L. rhamnosus in the amelioration of acne vulgaris has not been elucidated yet.MethodsTo mimic the human intestinal environment, a pseudo-germ-free rat model was used, and then gut microbiota from healthy individuals and acne patients were transplanted into rats. The effects of L. rhamnosus and tryptophan (Trp) metabolites on a rat acne model were investigated by gavage. Then, 16S rRNA analysis and targeted measurement of metabolites were performed to discover the differences in gut microbiota and metabolites between groups. Finally, HaCaT cells pretreated with Cutibacterium acnes were employed to validate the effect and mechanism of Trp metabolites on acne.ResultsL. rhamnosus significantly improved acne-like symptoms in rats by suppressing the level of inflammatory cytokines such as IL-1β, IL-6, and TNF-α. L. rhamnosus induced an increase in the production of indole-3-acetic acid (IAA) and indole via targeted Trp metabolic analyses. Furthermore, L. rhamnosus promoted bacterial diversity and also enhanced the Firmicutes/Bacteroidota (F/B) ratio, which was positively related to both IAA and indole. Finally, the roles of IAA and indole in alleviating acne vulgaris were confirmed both in vitro and in vivo, which could be reversed by AhR inhibitors.ConclusionOur study demonstrated that L. rhamnosus could exert its therapeutic effects on acne vulgaris by modulating the gut microbiota and regulating associated Trp metabolites.

Deoxynivalenol (DON) is a global contaminant found in crop residues, grains, feed, and animal and human food. Biodegradation is currently the best solution for addressing DON pollution. However, efficient detoxification bacteria or enzymes that can be applied in complex matrices are lacking. The aim of this study was to isolate a DON-detoxifying probiotic strain with a high degradation rate, a good safety profile, and a clear genetic background. One hundred and eight bacterial strains were isolated from 300 samples collected from a school farm and surrounding livestock farms. A new DON-degrading strain, Lactobacillus rhamnosus MY-1 (L. rhamnosus MY-1), with a degradation rate of 93.34% after 48 h and a comprehensive degradation method, was identified. Then, MY-1 at a concentration of 1 × 108 CFU/mL was administered to mice in a chronic intoxication experiment for 28 days. The experimental group showed significantly higher weight gain and exhibited good production performance compared to the control group. The length of the ileal villi in the experimental group was significantly longer than that in the control group. The expression of pro-inflammatory cytokines decreased, while the expression of anti-inflammatory factors increased in the experimental group. Whole-genome analysis revealed that most of the MY-1 genes were involved in carbohydrate metabolism and membrane transport, with a cluster of secondary metabolite genes encoding antimicrobial properties. In summary, this study successfully identified a Lactobacillus strain with good safety performance, high DON degradation efficiency, and a clear genetic background, providing a new approach for the treatment of DON contamination.

With the global increase in hyperglycemia and hyperlipidemia, there is an urgent need to explore dietary interventions targeting the inhibition of dipeptidyl peptidase-IV (DPP-IV) and lipid digestion and absorption. This study investigated how Lactobacillus rhamnosus GG (LGG) affects various aspects of black goji berry (BGB) (Lycium ruthenicum Murr.) juice, including changes in physicochemical and functional properties, as well as microbiological and sensory attributes. Throughout the fermentation process with 2.5–10% (w/v) BGB, significantly improved probiotic viability, lactic acid production, and decreased sugar content. While total flavonoids increase, anthocyanins decrease, with no discernible change in antioxidant activities. Metabolite profiling reveals elevated phenolic compounds post-fermentation. Regarding the inhibition of lipid digestion and absorption, fermented BGB exhibits improved bile acid binding, and disrupted cholesterol micellization by approximately threefold compared to non-fermented BGB, while also increasing pancreatic lipase inhibitory activity. Furthermore, a decrease in cholesterol uptake was observed in Caco-2 cells treated with fermented BGB (0.5 mg/mL), with a maximum reduction of 16.94%. Fermented BGB also shows more potent DPP-IV inhibition. Sensory attributes are significantly improved in fermented BGB samples. These findings highlight the potential of BGB as a bioactive resource and a promising non-dairy carrier for LGG, enhancing its anti-hyperglycemic and anti-hyperlipidemic properties.

Excessive dietary fat intake is closely associated with an increased risk of obesity, type 2 diabetes, cardiovascular disease, gastrointestinal diseases, and certain types of cancer. The administration of multi-strain probiotics has shown a significantly beneficial effect on the mitigation of obesity induced by high-fat diets (HFDs). In this study, Amuc_1100, an outer membrane protein of Akkermansia muciniphila, was fused with green fluorescent protein and LPXTG motif anchor protein and displayed on the surface of Lactobacillus rhamnosus (pLR-GAA) and Lactobacillus plantarum (pLP-GAA), respectively. The localization of the fusion protein on the bacterial cell surface was confirmed via fluorescence microscopy and Western blotting. Both recombinant strains demonstrated the capacity to ameliorate hyperglycemia and decrease body weight gain in a dose-dependent manner. Moreover, daily oral supplementation of pLR-GAA or pLP-GAA suppressed the HFD-induced intestinal permeability by regulating the mRNA expressions of tight junction proteins and inflammatory cytokines, thereby reducing gut microbiota-derived lipopolysaccharide concentration in serum and mitigating damage to the gut, liver, and adipose tissue. Compared with Lactobacillus rhamnosus treatment, high-dose pLR-GAA restored the expression level of anti-inflammatory factor interleukin-10 in the intestine. In conclusion, our approach enables the maintenance of intestinal health through the use of recombinant probiotics with surface-displayed functional protein, providing a potential therapeutic strategy for HFD-induced obesity and associated metabolic comorbidities.

Vaginally administered postbiotics derived from Lactobacillus were recently demonstrated to be effective in alleviating bacterial vaginosis and increasing pregnancy rates. However, their potential effect on sperm quality has not been well investigated. This controlled in vitro study aimed to assess the dose- and time-dependent effects of postbiotics derived from Lactobacillus rhamnosus PB01 (DSM 14870) on sperm quality parameters. The experiment was conducted in vitro to eliminate potential confounding factors from the female reproductive tract and vaginal microbiota. Sperm samples from 18 healthy donors were subjected to analysis using Computer-Aided Sperm Analysis (CASA) in various concentrations of postbiotics and control mediums at baseline, 60 min, and 90 min of incubation. Results indicated that lower postbiotic concentration (PB5) did not adversely affect sperm motility, kinematic parameters, sperm DNA fragmentation, and normal morphology at any time. However, concentrations exceeding 15% demonstrated a reduction in progressively motile sperm and a negative correlation with non-progressively motile sperm at all time points. These findings underscore the importance of balancing postbiotic dosage to preserve sperm motility while realizing the postbiotics’ vaginal health benefits. Further research is warranted to understand the underlying mechanisms and refine practical applications in reproductive health.

The high prevalence of constipation after fracture surgery brings intolerable discomfort to patients on the one hand, and affects post-surgery nutrient absorption on the other hand, resulting in poor prognosis. Given the acknowledged probiotic properties of Lactobacillus rhamnosus, 100 fracture patients with post-surgery constipation were centrally enrolled and administered orally with L. rhamnosus JYLR-127 to assess the efficacy of probiotic-adjuvant therapy in alleviating post-fracture constipation symptoms. The results showed that L. rhamnosus JYLR-127 improved fecal properties, promoted gastrointestinal recovery, and relieved constipation symptoms, which were mainly achieved by elevating Firmicutes (p < 0.01) and descending Bacteroidetes (p < 0.001), hence remodeling the disrupted intestinal microecology. In addition, blood routine presented a decrease in C-reactive protein levels (p < 0.05) and an increase in platelet counts (p < 0.05) after probiotic supplementation, prompting the feasibility of L. rhamnosus JYLR-127 in anti-inflammation, anti-infection and hemorrhagic tendency prevention after fracture surgery. Our study to apply probiotics in ameliorating constipation after fracture surgery is expected to bless the bothered patients, and provide broader application scenarios for L. rhamnosus preparations.

Aging is a time-dependent complex biological process of organisms with gradual deterioration of the anatomical and physiological functions. The role of gut microbiota is inevitable in the aging process. Probiotic interventions improve gut homeostasis and support healthy aging by enhancing beneficial species and microbial biodiversity in older adults. The present preliminary clinical trial delves into the impact of an 8-week Lactobacillus rhamnosus intervention (10 × 109 CFU per day) on the glycaemic index, lipid profile, and microbiome of elderly subjects. Body weight, body fat, fasting blood glucose, total cholesterol, triglyceride, high-density lipoprotein, and low-density lipoprotein (LDL) are assessed at baseline (Week 0) and after treatment (Week 8) in placebo and probiotic groups. Gaussian regression analysis highlights a significant improvement in LDL cholesterol in the probiotic group (p = 0.045). Microbiome analysis reveals numeric changes in taxonomic abundance at various levels. At the phylum level, Proteobacteria increases its relative frequency (RF) from 14.79 ± 5.58 at baseline to 23.46 ± 8.02 at 8 weeks, though statistically insignificant (p = 0.100). Compared to the placebo group, probiotic supplementations significantly increased the proteobacteria abundance. Genus-level analysis indicates changes in the abundance of several microbes, including Escherichia-Shigella, Akkermansia, and Bacteroides, but only Butyricimonas showed a statistically significant level of reduction in its abundance. Probiotic supplementations significantly altered the Escherichia-Shigella and Sutterella abundance compared to the placebo group. At the species level, Bacteroides vulgatus substantially increases after probiotic treatment (p = 0.021). Alpha and beta diversity assessments depict subtle shifts in microbial composition. The study has limitations, including a small sample size, short study duration, single-strain probiotic use, and lack of long-term follow-up. Despite these constraints, the study provides valuable preliminary insights into the multifaceted impact of L. rhamnosus on elderly subjects. Further detailed studies are required to define the beneficial effect of L. rhamnosus on the health status of elderly subjects.

Caries and periodontitis are the most prevalent oral diseases worldwide. Major factors contributing to the development of these oral conditions include poor oral hygiene, dental biofilm formation, high carbohydrates diet, smoking, other systemic diseases, and genetic factors. Various preventive measures have been established to mitigate the risk of caries and periodontal disease development. The present review aims to discuss the role of the probiotics Lactobacillus rhamnosus and Lactobacillus plantarum in the prevention and treatment of caries and periodontal diseases. The study was conducted in accordance with PRISMA guidelines and was registered on PROSPERO. The search involved PubMed, Web of Science, and Scopus and considered the PICO format. Studies were screened by two reviewers independently, and disagreements were solved by consensus with a third reviewer. Data extraction included details about the type of probiotics, strains, and purpose of administration. A total of 15 RCTs were included, of which just 1 was about tooth cavities. Overall, 87% of the included studies were good-quality papers regarding the Jadad Scale. Several studies agreed on the potential of probiotics L. rhamnosus and L. plantarum, both alone and combined, to prevent and improve clinical outcomes in caries and periodontal treatments, weaker evidence is provided for the microbiological benefit.

With the development and progress of human society and the continuous enhancement of people's health awareness, lactic acid bacteria inhibitors as a natural biopreservative have an irreplaceable position, with the advantages of high efficiency, broad spectrum, stability, safety and other characteristics. This paper studied the antibacterial effect of 23 lactic acid bacteria; the strains with the best antibacterial effect was identified by 16S rDNA method as being Lactobacillus rhamnosus L57 and was applied to banana storage. Our study showed that Lactobacillus rhamnosus L57 has a broad spectrum, inhibiting Escherichia coli, Staphylococcus aureus, Vibrio alginolyticus and Bacillus nato. Excluding the interference of organic acids, hydrogen peroxide and extracellular polysaccharide, the salting out of ammonium sulfate preliminarily determined that the proteins or peptides are the antibacterial substances in the antibacterial agent. Physicochemical properties showed that the microorganism had good antibacterial activity after trypsin, pepsin and papain, with good tolerance to pH, temperature and UV light, and the pH optimal range of 2.0-4.0. The mixture of freeze-dried powder of L. rhamnosus L57, chitosan and natamycin was used for banana preservation, with the proportion of freeze-dried powder of 1.25%, chitosan 1.5% and natamycin 0.03%. The preservation period was extended by 4 days compared with the absence of either bacteriostatic agent.

In this study, L-lactate dehydrogenase (Lr-L-LDH) from Lactobacillus rhamnosus was investigated. The well-known Lr-L-LDH1 was taken as the control, differences between Lr-L-LDH1 and Lr-L-LDH2 annotated from the genome of L. rhamnosus were analyzed. The primary structure, basic properties, hydrophobicity, secondary structure of Lr-L-LDH1 and Lr-L-LDH2 were predicted and analyzed using online websites and professional software. Homology modeling of the tertiary structure, molecular docking of enzymes and substrates, phylogenetic analysis, in vitro cloning, expression, and enzyme activity assays were further studied. The results showed that, in comparison to Lr-L-LDH1, although Lr-L-LHD2 had similar molecular characteristics, secondary and tertiary structures, Lr-L-LHD2 exhibited differences: A shorter sequences, low amino acid identities (48.08%), and a different phylogenetic status. Lr-L-LHD2 also contained a catalytically active site, a highly conserved NAD+ binding site sequence (GXGXXG), a three-dimensional active pocket domain, was an NAD+ dependent tetrameric L-lactate dehydrogenase, and be activated by fructose 1,6-diphosphate in the cytoplasm for catalyzing the reduction of pyruvate to L-lactic acid. In vitro enzyme activity was significantly lower than Lr-L-LDH1 (P

In order to increase the yield of L-lactic acid and reduce the production cost of L-lactic acid, a strain of Lactobacillus rhamnosus AK-0779 with alcohol tolerance and high yield of L-lactic acid was screened and domesticated. Corn distiller's grains were used as the nitrogen source for the fermentation medium of strain AK-0779 instead of part of yeast powder. On the basis of single factor experiments, three factors and three levels response surface optimization experiments were carried out on the addition of glucose, yeast powder and corn distiller's grains. The results showed that the optimal fermentation medium was glucose 9.80%, corn distiller's grains 0.98%, yeast powder 1.72%, L-lactic acid yield 78.91 g/L and sugar acid conversion rate 80.52%. There was no significant difference in the yield of L-lactic acid compared with yeast powder, which was 82.36 g/L, indicating that corn distiller's grains could effectively replace part of yeast powder as the nitrogen source of fermentation medium and reduce the production cost of L-lactic acid.

ABSTRACTBackground With increasing knowledge about the gut – bone axis, more studies for treatments based on the regulation of postmenopausal osteoporosis by gut microbes are being conducted. Based on our previous work, this study was conducted to further investigate the therapeutic effects of Lactobacillus rhamnosus GG (LGG) on ovariectomized (OVX) model rats and the immunological and microecological mechanisms involved.Results We found a protective effect of LGG treatment in OVX rats through changes in bone microarchitecture, bone biomechanics, and CTX-I, PINP, Ca, and RANKL expression levels. LGG was more advantageous in promoting osteogenesis, which may be responsible for the alleviation of osteoporosis. Th17 cells were imbalanced with Treg cells in mediastinal lymph nodes and bone marrow, with RORγt and FOXP3 expression following a similar trend. TNF-α and IL-17 expression in colon and bone marrow increased, while TGF-β and IL-10 expression decreased; however, LGG treatment modulated these changes and improved the Th17/Treg balance significantly. Regarding the intestinal barrier, we found that LGG treatment ameliorated estrogen deficiency-induced inflammation and mucosal damage and increased the expression of GLP-2 R and tight junction proteins. Importantly, 16S rRNA sequencing showed a significant increase in the Firmicutes/Bacteroidetes ratio during estrogen deficiency. Dominant intestinal flora showed significant differences in composition; LGG treatment regulated the various genera that were imbalanced in OVX, along with modifying those that did not change significantly in other groups with respect to the intestinal barrier, inflammation development, and bile acid metabolism.Conclusions Overall, LGG ameliorated estrogen deficiency-induced osteoporosis by regulating the gut microbiome and intestinal barrier and stimulating Th17/Treg balance in gut and bone.

Background: Dendritic cells, (DCs) as one of the important immune cell populations, are responsible for the initiation, development, and control of acquired immune responses. Myeloid dendritic cells can be used as a vaccine for several autoimmune diseases and cancers. Tolerogenic probiotics with regulatory properties can affect the maturation and development of immature dendritic cells (IDC) into mature DCs with certain immunomodulatory effects.Objective: To assess the immunomodulatory effect of Lactobacillus rhamnosus and Lactobacillus delbrueckii, as two tolerogenic probiotics, in the differentiation and maturation of myeloid dendritic cells.Methods: The IDCs were derived from the healthy donors in GM-CSF and IL 4 medium. Mature DCs (MDC) were produced with L. delbrueckii, L. rhamnosus, and LPS from IDCs. Real-Time PCR and flow cytometry were used to confirm the DC maturation and to determine DC markers as well as IDO, IL10, and IL12 expression levels, respectively.Results: Probiotic-derived DCs showed a significant reduction in the level of HLA-DR (P≤0.05), CD86 (P≤0.05), CD80 (P≤0.001), CD83 (P≤0.001), and CD1a. Also, the expression of IDO (P≤0.001) and IL10 increased while IL12 expression decreased (P≤0.001).Conclusion: Our findings revealed that tolerogenic probiotics could induce regulatory DCs by reducing co-stimulatory molecules along with increasing the expression of IDO and IL10 during the differentiation process. Therefore, the induced regulatory DCs probably can be used in the treatment of various inflammatory diseases.

Abstract Background Salmonella typhimurium (S.T), as an important foodborne bacterial pathogen, can cause diarrhea and gastroenteritis in humans and animals. Numerous studies have confirmed that exopolysaccharides (EPSs) have various biological functions, but the mechanism through which EPSs improve the immunity of animals against the invasion of pathogenic bacteria is unclear. Here, we explored the protective effect of EPSs of Lactobacillus rhamnosus GG (LGG) on the S.T-infected intestine. Methods Mice received adequate food and drinking water for one week before the start of the experiment. After 7 d of prefeeding, 2×108 CFU/mL S.T solution and an equivalent volume of saline (control group) were given orally for 1 d. On the fourth day, the mice were treated with 0.5 mg/mL EPSs, 1.0 mg/mL EPSs, 2.0 mg/mL EPSs, or 2.0 mg/mL penicillin for 7 d. Finally, the body and relative organ weight, histological staining, and the levels of antioxidant enzyme activity and inflammatory cytokines were determined. Results The S.T-infected mice exhibited symptoms of decreased appetite, somnolence, diarrhea and flagging spirit. Treatment with EPSs and penicillin improved the weight loss of the mice, and the high dose of EPSs showed the best therapeutic effect. EPSs significantly ameliorated S.T-induced ileal injury in mice. High-dose EPSs were more effective than penicillin for alleviating ileal oxidative damage induced by S.T. The mRNA levels of inflammatory cytokines in the ileum of mice showed that the regulatory effects of EPSs on inflammatory cytokines were better than those of penicillin. EPSs could inhibit the expression and activation of key proteins of the TLR4/NF-κB/MAPK pathway and thereby suppress the level of S.T-induced ileal inflammation. Conclusions EPSs attenuate S.T-induced immune responses by inhibiting the expression of key proteins in the TLR4/NF-κB/MAPK signaling pathway. Moreover, EPSs could promote bacterial aggregation into clusters, which may be a potential strategy for reducing the bacterial invasion of intestinal epithelial cells.

Lactobacillus rhamnosus and fructooligosaccharides (FOS) have been widely studied so far. However, the effects of L. rhamnosus on the intestinal microecological environment at the species level and the effect of different proportions of FOS on L. rhamnosus colonization in different parts of mice intestine are still unclear. The study results indicated that the specific bands of enterobacterial repetitive intergenic consensus polymerase chain reaction (ERIC-PCR) in the L. rhamnosus (LR) group significantly increased at 7 days. Although the number of bands was similar to the natural recovery (NR) group, the brightness of few bands significantly enhanced in the later stage of recovery. Besides, Southern-blot maps showed strong signals, indicating that the ERIC-PCR fingerprint could accurately reflect the changes in the mouse gut microbiota diversity. Further, the high-throughput results confirmed that the Lactobacillus and Akkermansia had different changes at different periods, but all of them showed an upward trend, while the Klebsiella were inhibited, thereby maintaining the intestinal microecology balance. Moreover, FOS exerted a positive effect on L. rhamnosus colonization in the gut.

Introduction: Atopic dermatitis (AD) is a chronic, inflammatory skin disorder that significantly impairs life quality and mostly affects children. Although classical approaches of treatment have been effective in mitigating the symptoms of AD, novel remedial measures based on a better understanding of immunopathogenesis are needed to target the underlying etiology. Aims and Objectives: Probiotics improve the intestinal micro environment and restore the balance of immune responses, thereby, emerging as a novel therapeutic option in the management of AD. Taking into account the updated information available, this review aims to discuss the beneficial effects of probiotics on gut microbiota, their role in strengthening the epidermal barrier and immunological responses, and hence their clinical usage in AD. Results: Classically, AD has been understood as a complex interplay between genetic susceptibility, barrier dysfunction, immune dysregulation, and exogenous factors. Current research has demonstrated that AD is associated with changes in both skin and gut microflora. The “gut-skin” axis hypothesis and the interaction between them have been demonstrated in research. Patients with AD have demonstrated gut dysbiosis, which leads to immune dysregulation and can serve as a potential trigger for AD. Probiotics, by their ability to put right gut dysbiosis, have demonstrated modest efficacy in reducing disease activity in clinical studies. Conclusion: Although multiple randomized controlled trials and meta-analyses have demonstrated the utility of probiotics as adjuvant therapy in the management of AD, the need of the hour is to develop precise guidelines pertaining to their usage in AD.

Caragana korshinskii is a forage shrub species with high-protein content that has been extensively used to alleviate feed shortages for ruminants in northern China. Herein, we investigated the effects of Lactobacillus rhamnosus and Lactobacillus buchneri on the fermentation quality, aerobic stability, and microbiome composition and the predicted functional characteristics of C. korshinskii silage. C. korshinskii silages were inoculated with and without L. rhamnosus or L. buchneri. After 14 and 56 days of ensiling, the aerobic stability was determined. The results revealed that after 14 and 56 days of ensiling, L. rhamnosus- and L. buchneri-inoculated silage exhibited increased acetic acid and lactic acid contents, whereas the pH and 2,3-butanediol and butyric acid contents were decreased compared with those of the control silage. The control silages that were opened at 14 and 56 d, deteriorated during the aerobic stability test, whereas silages inoculated with L. rhamnosus and L. buchneri did not exhibit any aerobic deterioration. The control silage showed an increased Clostridium and Bacillus abundance, whereas Lactobacillus abundance decreased compared with L. rhamnosus- and L. buchneri-inoculated silages, following the 7 days of aerobic exposure. The fermentation parameters were associated with microbial communities, including Lactobacillus, Pedicoccus, Weissella, Clostridium, and Bacillus. Carbohydrate and amino acid metabolisms in the control silage decreased after 7 days of aerobic exposure compared with lactic acid bacteria-inoculated silages. To conclude, next-generation sequencing combined with 16S ribosomal RNA gene-predicted functional analyses might provide new information about the silage quality during fermentation and the aerobic stability.

Introduction: Aluminium (Al) is accumulated in the brain causing neurotoxicity and neurodegenerative disease like Alzheimer's disease (AD), multiple sclerosis, autism and epilepsy. Hence, attenuation of Al-induced neurotoxicity has become a “hot topic“ in looking for an intervention that slow down the progression of neurodegenerative diseases.Objective: Our study aims to introduce a new strategy for hampering aluminum chloride (AlCl3)-induced neurotoxicity using a combination of sesamol with the probiotic bacteria; Lactobacillus rhamnosus (L. rhamnosus) and also to test their possible ameliorative effects on AlCl3-induced hepatotoxicity.Methods: Sprague-Dawley male rats were randomly divided into five groups (n = 10/group) which are control, AlCl3, AlCl3 + Sesamol, AlCl3 + L. rhamnosus and AlCl3 + Sesamol + L. rhamnosus. We surveilled the behavioral, biochemical, and histopathological alterations centrally in the brain and peripherally in liver.Results: This work revealed that the combined therapy of sesamol and L. rhamnosus produced marked reduction in brain amyloid-β, p-tau, GSK-3β, inflammatory and apoptotic biomarkers, along with marked elevation in brain free β-catenin and Wnt3a, compared to AlCl3-intoxicated rats. Also, the combined therapy exerted pronounced reduction in hepatic expressions of JAK-2/STAT-3, inflammatory (TNF-α, IL-6, NF-κB), fibrotic (MMP-2, TIMP-1, α-SMA) and apoptotic markers, (caspase-3), together with marked elevation in hepatic PPAR-γ expression, compared to AlCl3 -intoxicated rats. Behavioral and histopathological assessments substantiated the efficiency of this combined regimen in halting the effect of neurotoxicity.Discussion: Probiotics can be used as an add-on therapy with sesamol ameliorate AlCl3 -mediated neurotoxicity and hepatotoxicity.

China is seeing an increasing prevalence of inflammatory bowel disease (IBD) . During the process of exploring treatments for IBD, gut microbiota has been found to be crucial to the development of IBD along with the discovery of microbial-host interactions, suggesting that modulating gut microbiota may be a new strategy for the treatment of IBD. Recent studies have shown that Lactobacillus rhamnosus GG (LGG) , one most widely used probiotic strain, can achieve IBD alleviation by regulating intestinal microbial composition, maintaining intestinal epithelial barrier integrity, modulating intestinal immune response, inhibiting oxidative stress, and improving anxiety-depression status. Therefore, this paper presents a review of the mechanism of action of LGG in improving IBD, providing theoretical support for its clinical application.

As an alternative to chemicals and antibiotics, probiotics have proven to be effective in promoting successful aquaculture. They can improve water quality and generate high-quality livestock. The current study aimed to investigate the effect of adding Lactobacillus rhamnosus PTCC 1637 to the rearing water on survival rate and water quality during spawning on juvenile stages of rainbow trout of Oncorhynchus mykiss. This study has been implemented in three groups (1: eyed eggs, 2: alevin, and 3: fry) and three treatments (1 × 106, 1 × 107 bacteria per mL of water, and a control group). The abovementioned bacterial concentrations in group 1 were added on the days 1, 3, and 5; in group 2, every other day; and in group 3, from the days 1 to 20 on every other day, and from 20 to 60 once every four days. The results showed that the direct addition of L. rhamnosus to the rearing water improved the water quality parameters (DO, pH, ammonia and ammonium; p

[1]Introduction: Nowadays, with the development of probiotic products on the world market, the need for developing new products containing probiotic bacteria becomes more apparent. Probiotics are defined as living microorganisms that, if consumed in sufficient quantities, will have beneficial effects on the health of the host. Probiotics are now widely used in the production of food products and account for approximately 65% of functional foods. Probiotics often belong to either the genus Lactobacillus or Bifidobacterium. Lactobacillus rhamnosus is one of the known probiotic bacteria with beneficial properties. Prebiotics are defined as indigestible compounds, mainly carbohydrates that can be used as carbon source for probiotic bacteria and stimulate their growth and viability. Oligofructose is a type of short chain inulin and is one of the most well- known prebiotics. Moreover, microencapsulation of probiotic bacteria can improve the survival of these bacteria. In this approach, living probiotic cells are covered or trapped by various compounds. Hydrocolloids such as alginate and carbohydrates such as starch can be suitable compounds for microencapsulation. The purpose of this study was to investigate the effect of oligofructose and microencapsulation on the viability of Lactobacillus rhamnosus, textural, physicochemical and sensory characteristics of functional jelly. Materials and methods: In this study, different concentrations (0, 1.5 and 3 percent) of oligofructose as prebiotic were used to produce jelly samples, and 107 CFU/mL of probiotic bacteria (free and microencapsulated Lactobacillus rhamnosus) was inoculated. Microencapsulation of probiotic bacteria was performed by emulsion method using sodium alginate and corn resistant starch. The jelly samples were stored at 4˚C for two weeks. pH, acidity, dry matter, firmness, probiotic bacterial count and sensory properties (taste, odor, texture, color and overall acceptance) of the samples were evaluated on the first, 7th and 14th days of jelly production. Seven samples including 6 treatments and 1 control sample (without probiotic bacteria and prebiotic compound) with three replications were studied. The data were subjected to analysis of variance (ANOVA), followed by the Duncan’s multiple range test to determine the significant difference between samples at 95% confidence level (p0.05). Using 1.5% oligofructose and probiotic bacteria (free or microencapsulated) did not change the score of taste but the use of 3% oligofructose and free probiotic bacteria decreased the score of this parameter. The effect of storage time on sensory properties (taste, odor, texture, color and overall acceptance) was significant (p

In this study, five strains of Lactobacillus rhamnosus (L. rhamnosus) and two strains of Lactobacillus paracasei (L. paracasei) isolated from infants were used as experimental bacteria and Lactobacillus rhamnosus GG as control strain. The gastrointestinal tolerance of these strains were evaluated through acid resistance test, bile salt resistance test, hydrophobicity test, automatic aggregation ability test and cell adhesion test. L. rhamnosus SW-02, the best effective strain in tolerance evaluation, was co-cultured with intestinal organoids. The morphology and growth status of organoids were observed by inverted phase contrast microscope, and the budding rate was calculated. The proliferation status of organoids was evaluated by EdU staining. The mRNA levels of the proliferation marker Ki67, intestinal stem cell marker Lgr5 and tight junction protein Zo-1 were measured by real-time PCR. The level of MUC2 was detected by enzyme-linked immunosorbent assay. The results showed that compared with other strains, L. rhamnosus SW-02 had strong acid resistance. SW-02, SW-03 and SW-X had strong bile salt resistance. LGG, SW-01, SW-02, SW-04 and TX-02 had high hydrophobicity to xylene. For automatic aggregation capability, SW-01 was the strongest, followed by SW-02. The adhesion test of HT-29 cells showed that SW-01, SW-02 and SW-03 had high adhesion ability. When SW-02 was co-cultured with intestinal organoids, SW-02 significantly (P

In order to explore the effect of the compound probiotic powder of Lactobacillus rhamnosus and Bifidobacterium lactis on the immunity of mice and its food safety. In this study, low, medium and high doses (1.00, 0.67, 0.34g/kg BW) of compound probiotic powder were orally administered to mice to observe the organ/weight ratio, cellular immune function, humoral immune function, monocyte-macrophage function, NK cell activity and other indicators, comprehensively evaluate the effect of compound probiotic powder on the immune function of mice for 30 d. In addition, rat acute toxicity test, genetic toxicity test (mouse bone marrow cell micronucleus test, mouse sperm deformity test, Ames test) and 30-day feeding test for rats were used for food safety research. The results showed that, compared with the negative control group, the high-dose group mice had significantly increased delayed-type hypersensitivity (DTH), antibody cell production (PFC), serum hemolysin levels, and the phagocytosis index of carbon clearance in mice (P0.05). The growth and development of rats were good, and there was no significant difference in each index compared with the solvent control group (P>0.05). Therefore, the compound probiotic powder of Lactobacillus rhamnosus and Bifidobacterium lactis could enhance immunity without acute toxicity, genetic toxicity and subacute toxicity, and had high edible safety.

In order to develop a cheap and efficient culture system of Lactobacillus rhamnosus, walnut pulp was used to replace the nitrogen source in MRS medium. Taking cell growth as the indicator, the cultivation system was optimized in aspects of walnut pulp particle size, solid-liquid ratio, dosage, lipid removing treatment, and the shaking speed during the cultivation of Lactobacillus rhamnosus. The antibacterial activity and antioxidant activity of cell-free culture supernatant were also tested in all conditions. The obtained cells were also subjected to selenium enrichment treatment and then fed to mice for 3 d before they were exposed to simulated low-pressure hypoxia condition (altitude 5000 m) for 6 h to evaluate their anti-hypoxia potential. As results, complete replacement of the nitrogen source (beef extract and peptone) of MRS medium with the lipid-free walnut pulp filtered by 120 mesh screen at the solid-liquid ratio of 1:10 (g/mL) could reduce the cost of MRS medium by 70% and increase the cell yield by 2.5 times (from 2.6×109 to 7.6×109 CFU/mL), but did not affect the antibacterial activity and antioxidant activity of the fermentation broth (P>0.05). Continuous oral administration of selenium enriched bacteria for 3 days could significantly increase the hemoglobin content of mice when they were exposed to low-pressure hypoxia for 6 h (P

Probiotics are reported to improve the nutrition, immunity, and health of fish. Nuclease can hydrolyze nucleic acids of probiotics to produce nucleotides. The present study investigated the effect of stabilized fermentation product of nuclease-treated Lactobacillus rhamnosus GCC-3 (GCC-3 NT) on growth, non-specific immunity, liver health, and gut microbiota of zebrafish (Danio rerio). Compared to the high-fat diet (HFD) group, GCC-3 NT did not affect the growth performance of zebrafish. However, GCC-3 NT treatment can significantly increase the lysozyme activity and the total antioxidant capacity of body surface mucus. In addition, dietary GCC-3 NT significantly reduced the content of hepatic triglycerides (TAG) in zebrafish while significantly increased the expression of acyl-coenzyme A oxidases 3 (ACOX3) and proliferator-activated receptor γ coactivator 1α (PGC1α) compared with the HFD group. The 16S rRNA gene sequencing showed that GCC-3 NT reduced the relative abundance of Actinobacteria while increased Firmicutes at the phylum level. The relative abundance of Rhodococcus was significantly decreased and Lactobacillus and Staphylococcus abundance were significantly increased in the GCC-3 NT group compared to the HFD group. Furthermore, PCoA analysis showed GCC-3 NT diet had a significant effect on the autochthonous microbiota compared to the HFD diet. Together, our results showed that nuclease-treated L. rhamnosus fermentation product can improve the immunity, liver health and gut microbiota of zebrafish, suggesting that it can be potentially used as a functional feed additive for aquaculture.

Lactobacillus rhamnosus (LBS) is a well-documented probiotic strain in oncology and has a pivotal role in clinical applications. Here, we have investigated the protective effect of Lactobacillus rhamnosus on intestinal mucositis induced by cisplatin (CP) and explored the underlying mechanisms targeting inflammatory proteins, as well as the histological changes in the intestinal tissue of mice, in addition, the bacterial strains that may be related to the health-enhancing properties. BALB/c mice were pre-treated with or without LBS via oral gavage, followed by mucositis induction with cisplatin. Our results revealed that the LBS-treated groups significantly attenuated proinflammatory cytokine levels (IL-1β, IL-6, and TNF-α) compared to the CP group. Furthermore, LBS mitigated the damaged tight junction integrity caused by CP via up-regulating the levels of claudin, occludin, ZO-1, and mucin-2 protein (MUC-2). Finally, the 16S rRNA fecal microbiome genomic analysis showed that LBS administration enhanced the growth of beneficial bacteria, i.e., Firmicutes and Lachnospiraceae, while the relative abundance of the opportunistic bacteria Bacteroides and Proteobacteria decreased. Collectively, LBS was found to beneficially modulate microbial composition structure and functions and enrich the ecological diversity in the gut.

To prepare Eucheuma spinosum sulfated polysaccharides with low viscosity, high solubility and high bioavailability, fermented Eucheuma spinosum sulfate polysaccharides (F-ESP) was prepared by fermenting Lactobacillus rhamnoides, a common probiotic bacteria in fermented food, response surface methodology was used to optimize the preparation process with the yield of crude polysaccharide as the index. Low temperature freeze-thaw Eucheuma spinosum sulfate polysaccharide (L-ESP) was used as control. After purification by DEAE-52 column chromatography, the contents of total sugar, reducing sugar, sulfate and other substances and monosaccharide composition were determined, and the chemical composition of F-ESP was analyzed. The physical properties of F-ESP were characterized by viscosity, solubility, molecular weight and microstructure. The functional group structure of F-ESP was analyzed by infrared spectroscopy. The antiallergic activity of F-ESP was evaluated by Rat Basophilic Leukemia-2H3 (RBL-2H3) degranulation assay. When the ratio of material to liquid was 1:70, the inoculation amount was 5%, and the fermentation time was 24 h, the yield of crude polysaccharide was the highest (41.70%±2.00%). The purified F-ESP-3 monosaccharide structure was mainly composed of galactose, and the total sugar content was 97.77%±1.10%. The sulfate content was 28.40%±1.40%. Compared with L-ESP-3, the reducing sugar content of F-ESP-3 (7.87%±0.09%) was significantly higher, the molecular weight (33.58 kD) was significantly decreased (P

Spray drying is a commonly used method in the food industry since it is cost-effective and highly flexible, but spray drying typically results in lower survival rate of starter cultures than freeze-drying. In this study, the influence of heat pre-treatment (46 ℃, 1 h) and oxidative pre-treatment (0.5 mmol/L H2O2, 1 h) on the spray dried powder of Lactobacillus rhamnosus hsryfm 1301 was investigated. The results showed that heat pre-treatment and oxidative pre-treatment could improve the survival rate. The survival rate of the untreated group was only 29%, while the survival rates of heat pre-treated and oxidative pre-treated cells were 98% and 76%, which increased by 3.38 and 2.62 folds, respectively. At the same time, the acid-resistant of the untreated bacterial powder was 0.8%, while the acid-resistant survival rates of heat pre-treated and oxidative pre-treated bacterial powder were 76% and 72%, which increased by 95.0 and 92.5 times, respectively. Moreover, the heat pre-treated and oxidative pre-treated bacterial powder had good storage ability in the environment of −20 ℃. The survival rate of heat pre-treated bacterial powder remained greater than 80% after stored at −20 ℃ for 4 months, and that of oxidative pre-treated bacterial powder was greater than 40%. It was suggested that the strengthened heat stress and oxidative stress tolerance of L. rhamnosus hsryfm 1301 by heat pre-treatment and oxidative pre-treatment were helpful to survive spray drying.

The development of biofunctional non-dairy beverages with probiotics and prebiotics is a challenge and a frontier goal prompted by current consumers demand of health promoting foods, in particular from those with dietary restrictions related to dairy products. This study aimed to evaluate the survival of two commercially available probiotic strains, Lactobacillus rhamnosus LGG® and Lactobacillus paracasei (Lactobacillus casei 431), in a commercial sterile purple pitaya juice during storage at 5 °C. Additionally, the prebiotic effect of inulin (6.25 mg/ml) on the survival of L. rhamnosus, the most promising strain, was assessed. Physicochemical parameters such as betalain and soluble solids contents, pH, and color were also monitored during storage. Populations of L. rhamnosus and L. paracasei decreased from 11 to 7 log CFU/ml after 12 ± 1 and 0.93 ± 0.05 d, respectively. No changes on the individual betalain contents, as well as for total betalain concentrations (28 ± 3 μg/ml) were observed after probiotic inoculation nor during storage for all experimental trials and regardless of the strain evaluated (P 0.05) influence the survival kinetics of L. rhamnosus in the juice during storage and the juice color, pH and soluble solids content remained unaffected as well. Purple pitaya juice represents therefore a promising non-dairy food matrix for probiotics delivery.

Gut microbiota plays a role in the neonatal immune education and could influence susceptibility to Th2-type immune disorders, such as allergies, the most prevalent chronic diseases in early childhood. We studied the impact of oral Lactobacillus rhamnosus (L.rhamnosus) supplementation to pregnant/breastfeeding C57BL/6 mice on the development of allergic airways disease in their offspring. We observed that mice, from L.rhamnosus-treated mothers, inoculated with ovalbumin (OVA)-Aluminium hydroxide (ALUM) at 3 days of life and challenged intranasally 4 weeks later showed decreased Th2-associated cytokines, IgE and IgG1, lung eosinophilia and airway hyper-reactivity compared to OVA-sensitized mice from untreated mothers. In that setting, the L.rhamnosus treatment increased the number and maturation of splenic neonatal type 1 conventional dendritic cells (cDC1) that remained largely dominant over the cDC2 and favored their OVA-specific Th1 differentiation. In response to inhaled house dust mite (HDM) allergen, the maternal L.rhamnosus supplementation increased the number of neonatal pulmonary cDC1 expressing lower amount of costimulatory molecules compared with no supplementation and decreased the number of cDC2 without affecting their costimulatory molecules expression. An HDM-specific Foxp3+RORγt+ Treg polarization was monitored in the lung draining lymph nodes. Finally, we confirmed the inhibitory effect of maternal L.rhamnosus treatment on all the measured features of the HDM allergic airways reaction in their offspring. We conclude that maternal L.rhamnosus administration prevents Th2-type allergic airways disease in their neonates by favoring splenic cDC1/Th1 responses against ALUM-adjuvanted OVA or by promoting a pulmonary Foxp3+RORγt+ Treg activation against inhaled HDM.

There are many studies devoted to the study of the intestinal microbiota. They prove the leading role of microorganisms in the formation of the immune response of the host. Intestinal microbiota contributes to the switch of preferential differentiation of Th-lymphocytes from Th2-type, typical for newborns, to the formation of Tr-cells (regulatory) with a corresponding increase in the formation of TGF-β and IL-10. Consequently, immunological tolerance is formed under the influence of the intestinal microbiota. This process occurs in the first months of life and has a long-term effect, setting the features of the body’s immune response to external infectious and food antigens, which determines the predisposition to the development of food allergies later in life. This article describes the factors that influence the colonization of the baby’s gut. Maternal gut microbiota, maternal health, mode of delivery (natural childbirth or cesarean section), type of feeding (breastfeeding or formula feeding) - all these factors affect the colonization of the gut and the formation of an immune response. In addition, lactobacilli and bifidobacteria play a critical role in the formation of immunological tolerance. Probiotics can modulate the immune response similar to the intestinal microbiota. Lactobacillus and bifidobacterium infant strains are optimal. Clinical studies have proven the effectiveness of perinatal and early administration of probiotics for the prevention of food allergies. A balanced combination of Lactobacillus rhamnosus and Bifidobacterium longum is a symbiosis of bacteria that enhances the effectiveness of each other, which promotes the optimal establishment of the intestinal microbiocenosis from birth and is the prevention of both infectious diseases and food allergies. A combination probiotic containing both Lactobacillus rhamnosus and Bifidobacterium longum seems to be the optimal choice.

Antimicrobial resistance is one of the biggest health challenges nowadays. Probiotics are promising candidates as feed additives contributing to the health of the gastrointestinal tract. The beneficial effect of probiotics is species/strain specific; the potential benefits need to be individually assessed for each probiotic strain or species. We established a co-culture model, in which gastrointestinal infection was modeled using Escherichia coli (E. coli) and Salmonella enterica serovar Typhimurium (S. enterica serovar Typhimurium). Using intestinal porcine epithelial cells (IPEC-J2), the effects of pre-, co-, and post-treatment with Lactobacillus (L.) rhamnosus on the barrier function, intracellular (IC) reactive oxygen species (ROS) production, proinflammatory cytokine (IL-6 and IL-8) response, and adhesion inhibition were tested. E. coli- and S. Typhimurium-induced barrier impairment and increased ROS production could be counteracted using L. rhamnosus (p < 0.01). S. Typhimurium-induced IL-6 production was reduced via pre-treatment (p < 0.05) and post-treatment (p < 0.01); increased IL-8 secretion was decreased via pre-, co-, and post-treatment (p < 0.01) with L. rhamnosus. L. rhamnosus demonstrated significant inhibition of adhesion for both S. Typhimurium (p < 0.001) and E. coli (p < 0.001 in both pre-treatment and post-treatment; p < 0.05 in co-treatment). This study makes a substantial contribution to the understanding of the specific benefits of L. rhamnosus. Our findings can serve as a basis for further in vivo studies carried out in pigs and humans.

Byung Chull An,1 Yeo-Sang Yoon,1 Ho Jin Park,1 Sangkyun Park,1 Tai Yeub Kim,1 Jun Young Ahn,1 Daebeom Kwon,1 Oksik Choi,1 Jin Young Heo,1 Yongku Ryu,1 Joong-Hyun Kim,2 Heejong Eom,2 Myung Jun Chung1 1R&D Center, Cell Biotech, Co., Ltd., Gimpo-si, Gyeonggi-do, Korea; 2Laboratory Animal Center, Osong Medical Innovation Foundation, Chungbuk, Cheongju, 28160, KoreaCorrespondence: Myung Jun Chung Tel +82-31-987-6205Fax +82-31-987-6216Email ceo@cellbiotech.comPurpose: This study aimed to toxicological evaluate a probiotics-based delivery system for p8 protein as an anti-colorectal cancer drug.Introduction: Lactic acid bacteria (LAB) have been widely ingested for many years and are regarded as very safe. Recently, a Pediococcus pentosaceus SL4 (PP) strain that secretes the probiotic-derived anti-cancer protein P8 (PP-P8) has been developed as an anti-colorectal cancer (CRC) biologic by Cell Biotech. We initially identified a Lactobacillus rhamnosus (LR)-derived anti-cancer protein, P8, that suppresses CRC growth. We also showed that P8 penetrates specifically into CRC cells (DLD-1 cells) through endocytosis. We then confirmed the efficacy of PP-P8, showing that oral administration of this agent significantly decreased tumor mass (∼ 42%) relative to controls in a mouse CRC xenograft model. In terms of molecular mechanism, PP-P8 induces cell-cycle arrest in G2 phase through down-regulation of Cyclin B1 and Cdk1. In this study, we performed in vivo toxicology profiling to obtain evidence that PP-P8 is safe, with the goal of receiving approval for an investigational new drug application (IND).Methods: Based on gene therapy guidelines of the Ministry of Food and Drug Safety (MFDS) of Korea, the potential undesirable effects of PP-P8 had to be investigated in intact small rodent or marmoset models prior to first-in-human (FIH) administration. The estimated doses of PP-P8 for FIH are 1.0× 1010 – 1.0× 1011 CFU/person (60 kg). Therefore, to perform toxicological investigations in non-clinical animal models, we orally administered PP-P8 at doses of 3.375 × 1011, 6.75 × 1011, and 13.5× 1011 CFU/kg/day; thus the maximum dose was 800– 8000-fold higher than the estimated dose for FIH.Results: In our animal models, we observed no adverse effects of PP-P8 on clinicopathologic findings, relative organ weight, or tissue pathology. In addition, we observed no inflammation or ulceration during pathological necropsy.Conclusion: These non-clinical toxicology studies could be used to furnish valuable data for the safety certification of PP-P8.Keywords: biologics, therapeutic protein, probiotics derived anti-cancer protein P8, Pediococcus pentosaceus SL4, Lactobacillus rhamnosus, colorectal cancer, drug delivery system, protein secretion system, oral administration, non-clinical trial, toxicology finding

Objective(s): The present in vitro study aimed to evaluate whether Lactobacillus delbrueckii and Lactobacillus rhamnosus treatments can induce regulatory phenotype, together with modulating the expression of chemokine receptors (CRs) in dendritic cells (DCs). The CRs of DCs have been found to be involved in the pathogenesis of Systemic lupus erythematosus (SLE) through directing recruitment and migration of immune cells.Materials and Methods: In brief, monocytes of patients with SLE and healthy donors were isolated and differentiated to regulatory or inflammatory mature DCs through treatment with L. delbrueckii, L. rhamnosus, mixed probiotics, and LPS. Results: FACScan analysis showed that the expression of CRs including CXCR3, CCR5, CCR4, and CCR3, was significantly reduced in all probiotic-treated groups of SLE and healthy (control) donors when compared with the LPS treated group. Conclusion: The results demonstrated that tolerogenic probiotics could prevent or decrease the expression of inflammatory CRs on the surface of tolerogenic DCs during the maturation process.

Psychological stress negatively affects the intestinal barrier function in animals and humans. We aimed to study the effect of Lactobacillus rhamnosus CNCM I-3690 on intestinal permeability and stress-markers during public speech. Healthy students were randomized to L. rhamnosus-containing (test) or acidified (placebo) milk consumed twice daily for 4 weeks, with 46 subjects per treatment group. Small intestinal permeability was quantified by a 2 h urinary lactulose–mannitol ratio (LMR, primary outcome), fractional excretion of lactulose (FEL) and mannitol (FEM). Salivary cortisol, State-Trait Anxiety Inventory (STAI) and Perceived Stress scores (PSS) were collected. No between-treatment differences were found for LMR (p = .71), FEL or FEM. Within-treatment analyses showed similar LMR and FEL but a stress-induced increase of FEM with the placebo (p

Abstract Objective Type 2 diabetes mellitus (T2DM) is still a challenge for physicians to manage patient’s circumstances. It is assumed that alterations in the normal flora may be involved in the pathogenesis of T2DM through inducing chronic inflammation. To investigate the effect of Lactobacillus rhamnosus as a common probiotic on T2DM, we induced an experimental model of T2DM in adult male Zebrafish by gradient hyper-glucose accumulation methodology. Results In this trial 3-month old male adult Zebrafish were divided in to four groups including two control groups and T2DM induced groups with or without probiotic treatment. After 5 days of acclimation, T2DM was induced by a gradient hyper-glucose accumulation methodology. Diabetic fishes had statistically abnormal blood glucose and pro-inflammatory cytokine levels compared to control group (p = 0.0001). These results suggest that probiotic intervention decreased the blood glucose level in the T2DM-P group by decreasing pro-inflammatory cytokines responsible for signaling in T2DM therapeutic modalities.