Search Results (17)

The development of bakeable foods supplemented with probiotics requires novel strategies to preserve the functionality of probiotic cells during thermal and gastrointestinal stress conditions. The objective of the present study was to evaluate the protective effect of multilayer double emulsions (W₁/O/W₂) stabilized with pectin-protein complexes on the viability of Limosilactobacillus reuteri (Lr) under thermal treatment (95 °C, 30 min), storage (4 °C, 28 d), and simulated gastrointestinal conditions. Emulsions were prepared with whey protein isolate (WPI) or sodium caseinate (Cas) as outer aqueous phase emulsifiers, followed by pectin coating and ionic gelation with calcium. All emulsions were stable and exhibited high encapsulation efficiency (>92%) with initial viable counts of 9 log CFU/mL. Double emulsions coated with ionically gelled pectin showed the highest protection against heat stress and gastrointestinal conditions due to the formation of a denser layer with lower permeability, regardless of the type of protein used as an emulsifier. At the end of storage, Lr viability exceeded 7 log CFU/mL in cross-linked pectin-coated microcapsules. These microcapsules maintained >6 log CFU/mL after thermal treatment, while viability remained >6.5 log CFU/mL during digestion and >5.0 log CFU/mL after consecutive heat treatment and simulated digestion. According to these results, the combination of double emulsion, multilayer formation and ionic crosslinking emerges as a promising microencapsulation technique. This approach offers enhanced protection for probiotics against extreme thermal and digestive conditions compared to previous studies that only use double emulsions. These findings support the potential application of this encapsulation method for the formulation of functional bakeable products. Full article

Lactic acid bacteria play a crucial role in maintaining the health of the host’s gut microbiota. In this study, the anti-inflammatory properties of Limosilactobacillus reuteri LR20-6 and Lacticplantibacillus plantarum L272 were evaluated using a mouse model of diarrhea induced by Escherichia coli. We also investigated their effects on gut microbiota regulation. The results indicated that both Lacticplantibacillus plantarum and Limosilactobacillus reuteri could reduce inflammation by inhibiting the expression of inflammatory factors IL-6 and TNF-α and blocking the MyD88 and NF-kB/p65 signaling pathways. Additionally, after intervention with these strains, the relative abundance of Lactobacillus was significantly increased. This suggested that Lacticplantibacillus plantarum and Limosilactobacillus reuteri could mitigate the severity of E. coli-induced diarrhea and enhance the abundance of beneficial probiotics in the gut of animals. Full article

The underexplored fruit from the juçara palm tree (Euterpe edulis Martius) has bioactive compounds with antioxidant activities, such as phenolic acids and anthocyanins. This fruit’s pulp presents itself as an appropriate fermentation medium for probiotic bacteria growth. Therefore, this study was conducted to evaluate the effects of fermentation by Limosilactobacillus reuteri LR92 (JLR) and Bifidobacterium animalis ssp. lactis BB-12 (JBB) on the bioactive compound contents of the juçara pulp, before and after a gastrointestinal simulation. The pulp of the juçara fruit showed probiotic counts of 8.70 ± 0.07 log UFC/mL for JLR and 8.44 ± 0.09 log UFC/mL for JBB, after 24 h of fermentation. Fermentation with the strains used modified the proportions of fatty acids (fatty acids esters were quantified using a gas chromatography equipment) and fibers when compared to the non-fermented pulp. The antioxidant capacity determined by DPPH, FRAP and ABTS showed significant reduction after the gastrointestinal simulation for samples. Phenolic compound analysis by UPLC-MS/MS showed, after fermentation, a greater amount of ferulic, protocatechuic and catechin acids in the samples. These results show changes in the bioactive compounds due to the fermentation of the juçara pulp by probiotics. However, these compounds showed bioactive potential and were bioaccessible after the gastrointestinal simulation, with the pulp being a potential means for bacteria growth, which may bring health benefits. Full article

Fermented milk with probiotic bacteria is a functional food, and adding fruit can enhance its taste. Juçara, the fruit of the Euterpe edulis Martius palm tree, is known for its natural antioxidant properties. This study aimed to assess the antioxidant capacity of milk fermented by Limosilactobacillus reuteri LR92 with juçara pulp (JFM) over 30 days of storage at 4 °C and its protective effect on probiotic cells against reactive oxygen species (ROS). Phenolic compounds and antioxidant activities were measured using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and ferric-reducing antioxidant power (FRAP) assays during storage. The resistance of L. reuteri to hydrogen peroxide, superoxide anions, and hydroxyl radicals was also tested. The results indicated that JFM maintained stability in its composition, except for color, which showed reduced brightness by the end of the 30 days. Although antioxidant activity measured by DPPH and FRAP decreased (83.92–67.03 µmol TEAC.g⁻¹ and 1185.64–830 g TEAC.100 g.mL⁻¹, respectively), it remained higher than the control (21.90–24.50 µmol TEAC.g⁻¹ and 235.77–229.87 g TEAC.100 g.mL⁻¹, respectively). Phenolic content remained consistent. In addition, juçara pulp significantly protected L. reuteri cells from ROS. Therefore, juçara-enriched fermented milk not only improved antioxidant properties but also shielded probiotics from oxidative stress, highlighting its potential as a functional food with added health benefits. Full article

Background/Objectives: Limosilactobacillus (Lm.) reuteri is a widely utilized probiotic, recognized for its significant role in alleviating symptoms associated with gastrointestinal and psychiatric disorders. However, the effectiveness of Lm. reuteri is strain-specific, and its genetic diversity leads to significant differences in phenotypes among different strains. This study aims to identify potential probiotic strains by comparing the strain-specific characteristics of Lm. reuteri to better understand their efficacy and mechanisms in alleviating stress-induced anxiety-like behaviors and gastrointestinal symptoms. Methods: We cultivated 11 strains of Lm. reuteri from healthy human samples and conducted phenotypic and genomic characterizations. Two strains, WLR01 (=GOLDGUT-LR99) and WLR06, were screened as potential probiotics and were tested for their efficacy in alleviating anxiety-like behavior and intestinal symptoms in mouse models subjected to sleep deprivation (SD) and water avoidance stress (WAS). Results: The results showed that the selected strains effectively improved mouse behaviors, including cognitive impairment and inflammatory response, as well as improving anxiety and regulating gut microbiota composition. The improvements with WLR01 were associated with the regulation of the NLRP3 inflammasome pathway in the SD model mice and were associated with visceral hypersensitivity and intestinal integrity in the WAS model mice. Conclusions: In summary, this study identified the Lm. reuteri strain WLR01 as having the potential to alleviate anxiety-like behavior and intestinal symptoms through the analysis of Lm. reuteri genotypes and phenotypes, as well as validation in mouse models, thereby laying the foundation for future clinical applications. Full article

Despite recent advances in cancer therapy, ovarian cancer remains the most lethal gynecological cancer worldwide, making it crucial and of the utmost importance to establish novel therapeutic strategies. Adjuvant radiotherapy has been assessed historically, but its use was limited by intestinal toxicity. We recently established the role of Limosilactobacillus reuteri in releasing IL-22 (LR-IL-22) as an effective radiation mitigator, and we have now assessed its effect in an ovarian cancer mouse model. We hypothesized that an LR-IL-22 gavage would enable intestinal radioprotection by modifying the tumor microenvironment and, subsequently, improving overall survival in female C57BL/6MUC-1 mice with widespread abdominal syngeneic 2F8cis ovarian cancer. Herein, we report that the LR-IL-22 gavage not only improved overall survival in mice when combined with a PD-L1 inhibitor by inducing differential gene expression in irradiated stem cells but also induced PD-L1 protein expression in ovarian cancer cells and mobilized CD8+ T cells in whole abdomen irradiated mice. The addition of LR-IL-22 to a combined treatment modality with fractionated whole abdomen radiation (WAI) and systemic chemotherapy and immunotherapy regimens can facilitate a safe and effective protocol to reduce tumor burden, increase survival, and improve the quality of life of a locally advanced ovarian cancer patient. Full article

Ribonucleoside hydrolase C (RihC, EC 3.2.2.1, 3.2.2.2, 3.2.2.3, 3.2.2.7, 3.2.2.8) belongs to the family of ribonucleoside hydrolases Rih and catalyzes the cleavage of ribonucleosides to nitrogenous bases and ribose. RihC is one of the enzymes that are synthesized by lactobacilli in response to the presence of Klebsiella. To characterize this protein from Limosilactobacillus reuteri LR1, we cloned and expressed it. The activity of the enzyme was studied towards a wide range of substrates, including ribonucleosides, deoxyribonucleosides as well as an arabinoside. It was shown that the enzyme is active only with ribonucleosides and arabinoside, with the best substrate being uridine. The thermal stability of this enzyme was studied, and its crystal structure was obtained, which demonstrated the tetrameric architecture of the enzyme and allowed to shed light on a correlation between its structure and enzymatic activity. Comprehensive comparisons of all known RihC structures, both existing crystal structures and computed model structures from various species, were made, allowing for the identification of structural motifs important for enzyme functioning. Full article

Salmonella enterica serovar Typhimurium (S. typhimurium) is an important foodborne pathogen that infects both humans and animals and develops acute gastroenteritis. As porcine intestines are relatively similar to the human ones due to their relatively similar sizes and structural similarity, S. typhimurium causes analogous symptoms in both. Novel strategies for controlling S. typhimurium infection are also desired, such as mucosal-targeted delivery of probiotics and antimicrobial peptides. The bovine lactoferricin-lactoferrampin-encoding Limosilactobacillus reuteri (LR-LFCA) strain improves intestinal barrier function by strengthening the intestinal barrier. Weaned piglets were selected for oral administration of microencapsulated LR-LFCA (microcapsules entrap LR-LFCA into gastro-resistant polymers) and then infected with S. typhimurium for 3 days. We found that orally administering microencapsulated LR-LFCA to weaned piglets attenuated S. typhimurium-induced production of inflammatory factors in the intestinal mucosa by inhibiting the nuclear factor-kappa B (NF-κB) and P38 mitogen-activated protein kinases (MAPK) signaling pathway. Moreover, microencapsulated LR-LFCA administration significantly suppressed the oxidative stress that may correlate with gut microbiota (reduced Salmonella population and increased α-diversity and Lactobacillus abundance) and intestinal function (membrane transport and metabolism). Our work demonstrated that microencapsulated LR-LFCA effectively targeted intestine delivery of Lactobacillus and antimicrobial peptides and modulated gut microbiota and mucosal immunity. This study reveals a novel targeting mucosal strategy against S. typhimurium infection. Full article

The multifunctional antibacterial peptide lactoferricin-lactoferrampin (LFCA) is derived from bovine lactoferrin. Optimization of the fermentation process should be studied since different microorganisms have their own favorable conditions and processes for growth and the production of metabolites. In this study, the culture conditions of a recombinant strain, pPG-LFCA-E/LR-CO21 (LR-LFCA), expressing LFCA was optimized, utilizing the high-density fermentation process to augment the biomass of LimosiLactobacillus reuteri and the expression of LFCA. Furthermore, an assessment of the protective effect of LR-LFCA on intestinal inflammation induced by lipopolysaccharide (LPS) was conducted to evaluate the impact of LR-LFCA on the disease resistance of piglets. The findings of this study indicate that LR-LFCA fermentation conditions optimally include 2% inoculation volume, 36.5 °C fermentation temperature, 9% dissolved oxygen concentration, 200 revolutions/minute stirring speed, pH 6, 10 mL/h glucose flow, and 50% glucose concentration. The inclusion of fermented LR-LFCA in the diet resulted in an elevation of immunoglobulin levels, significant upregulation of tight junction proteins ZO-1 and occludin, reinforcement of the intestinal barrier function, and significant amelioration of the aberrant alterations in blood physiological parameters induced by LPS. These results offer a theoretical framework for the implementation of this micro-ecological preparation in the field of piglet production to enhance intestinal well-being. Full article

Little is known about how combining probiotics affects the storage survival and functional performance of individual probiotics when incorporated into non-dairy drinks. Viability of Lacticaseibacillus rhamnosus GG (LG), Limosilactobacillus reuteri ATCC 55730 (LR), Bifidobacterium animalis subsp. lactis BB-12 (Bb), and Propionibacterium jensenii 702 (PJ), either alone or in multi-species combinations included in orange juice (OJ), were assessed during storage in refrigerated conditions and compared with bottled water (BW). The tolerance of probiotics included in refrigerated OJ to simulated gastrointestinal conditions was also examined. LG and LR viabilities were significantly higher in OJ than in BW (p ≤ 0.001), while the reverse was evident for PJ. Bb maintained high viability in both drinks. LG-PJ in both drinks and Bb-PJ in BW resulted in greater viabilities among the paired combinations compared to their respective monocultures when incorporated separately (p ≤ 0.001). The viability of LG in the LG-Bb-PJ combination improved significantly in BW compared with LG alone (p ≤ 0.001). OJ did not alter bacterial tolerance to simulated gastric juice but diminished tolerance to simulated intestinal juice (SIJ). In all combinations, tolerance of LG and LR to SIJ was improved, whereas tolerance of PJ declined significantly compared with respective monocultures (p ≤ 0.001). In conclusion, probiotic storage stability and gastrointestinal transit tolerance were species-dependent and affected by carrier type and combinations. These effects should be considered when formulating probiotic products. Full article

Irradiation can be an effective treatment for ovarian cancer, but its use is limited by intestinal toxicity. Thus, strategies to mitigate toxicity are important and can revitalize the current standard of care. We previously established that LR-IL-22 protects the intestine from WAI. We now hypothesize that LR-IFN-β is an effective radiation protector and mitigator and is rapidly cleared from the digestive tract, making it an option for intestinal radioprotection. We report that the gavage of LR-IFN-β during WAI provides improved intestinal barrier integrity and significantly preserves the numbers of Lgr5+GFP+ intestinal stem cells, improving survival. The rapid clearance of the genetically engineered probiotic from the digestive tract renders it a safe and feasible radiation mitigator. Therefore, the above genetically engineered probiotic is both a feasible and effective radiation mitigator that could potentially revolutionize the management of OC patients. Furthermore, the subsequent addition of platinum/taxane-based chemotherapy to the combination of WAI and LR-IFN-β should reduce tumor volume while protecting the intestine and should improve the overall survival in OC patients. Full article

Limosilactobacillus (L.; previously Lactobacillus) reuteri has been shown to influence gastrointestinal (GI) tolerance. This study was a secondary analysis of GI tolerance data from a multi-country, cross-sectional, observational study in healthy infants using the validated Infant Gastrointestinal Symptom Questionnaire (IGSQ) and a gut comfort questionnaire. Breastfed infants (BFI; n = 760) were compared to formula-fed infants receiving either L. reuteri-containing formula (FFI + LR; n = 470) or standard formula without any probiotic or prebiotic (FFI-Std; n = 501). The IGSQ composite scores (adjusted mean ± SE) in FFI + LR (22.17 ± 0.39) was significantly lower than in FFI-Std (23.41 ± 0.37) and similar to BFI (22.34 ± 0.30;), indicating better GI tolerance in FFI + LR than in FFI-Std. Compared with FFI-Std, FFI + LR had lower reports of difficulty in passing stools (11% vs. 22%; adjusted-odds ratio (OR) (95%CI) = 0.46 (0.31–0.68)), fewer hard stools (mean difference = −0.12 (−0.21, −0.02)) and less physician-confirmed colic (OR = 0.61 (0.45–0.82)), and similar to BFI. Parent-reported crying time (mean difference = −0.15 (−0.28, −0.01)), frequency of spitting-up/vomiting (mean difference = −0.18 (−0.34, −0.03)), volume of spit-up (mean difference = −0.20 (−0.32, −0.08)) and fussiness due to spitting-up/vomiting (mean difference = −0.17 (−0.29, −0.05)) were lower in FFI + LR versus FFI-Std and similar to BFI. In this study, L. reuteri-containing formula was associated with improved digestive tolerance and behavioral patterns. Full article

The human oral pathobionts Aggregatibacter actinomycetemcomitans, Streptococcus mitis and Streptococcus mutans, in dysbiosis-promoting conditions, lead to oral infections, which also represent a threat to human systemic health. This scenario may be worsened by antibiotic misuse, which favours multi-drug resistance, making the research on pathogen containment strategies more than crucial. Therefore, we aimed to in vitro select the most promising probiotic strains against oral pathogen growth, viability, biofilm formation, and co-aggregation capacity, employing both the viable probiotics and their cell-free supernatants (CFSs). Interestingly, we also assessed probiotic efficacy against the three-pathogen co-culture, mimicking an environment similar to that in vivo. Overall, the results showed that Lactobacillus CFSs performed better than the Bifidobacterium, highlighting Limosilactobacillus reuteri LRE11, Lacticaseibacillus rhamnosus LR04, Lacticaseibacillus casei LC04, and Limosilactobacillus fermentum LF26 as the most effective strains, opening the chance to deeper investigation of their action and CFS composition. Altogether, the methodologies presented in this study can be used for probiotic efficacy screenings, in order to better focus the research on a viable probiotic, or on its postbiotics, suitable in case of infections. Full article

Soybean protein and peptides have the potential to promote the growth of Lactobacillus, but the mechanisms involved are not well understood. The purpose of this study is to investigate differentially expressed genes (DEGs) of Limosilactobacillus reuteri (L. reuteri) LR08 responding to soybean protein and peptides using transcriptome. The results showed that both digested protein (dpro) and digested peptides (dpep) could enhance a purine biosynthesis pathway which could provide more nucleic acid and ATP for bacteria growth. Moreover, dpep could be used instead of dpro to promote the ABC transporters, especially the genes involved in the transportation of various amino acids. Interestingly, dpro and dpep played opposite roles in modulating DEGs from the acc and fab gene families which participate in fatty acid biosynthesis. These not only provide a new direction for developing nitrogen-sourced prebiotics in the food industry but could also help us to understand the fundamental mechanism of the effects of dpro and dpep on their growth and metabolisms and provides relevant evidence for further investigation. Full article

A number of studies have confirmed the relationship between constipation and gut microbiota. Additionally, many human and animal experiments have identified probiotics as effectors for the relief of constipation symptoms. In this study, probiotic compounds, including Lactobacillus acidophilus LA11-Onlly, Lacticaseibacillus rhamnosus LR22, Limosilactobacillus reuteri LE16, Lactiplantibacillus plantarum LP-Onlly, and Bifidobacterium animalis subsp. lactis BI516, were administered to mice with loperamide-induced constipation, and the impacts of these strains on constipation-related indicators and gut microbiota were evaluated. The effects of probiotic compounds on constipation relief were associated with various aspects, including gastrointestinal transit rate, number and weight of stools, serum and intestinal gastrointestinal regulatory hormones, and serum cytokines. Some of the probiotic compounds, including Limosilactobacillus reuteri, Lactiplantibacillus plantarum, and Lacticaseibacillus rhamnosus, were found to colonize the intestinal tract. Furthermore, higher dosages promoted the colonization of specific strains. This study yields a new perspective for the clinical use of probiotics to improve constipation symptoms by combining strains with different mechanisms for the alleviation of constipation. Full article