Investigation of impact of siderophore and process variables on production of iron enriched Saccharomyces boulardii by Plackett-Burman design.

The primary cause of anemia worldwide is due to poor diet and iron deficiency. Iron (Fe) enriched yeast can be the most effective way to manage anemia because of the capability for biotransformation of mineral to organic and bioavailable iron. To overcome the low richness of yeast, the use of siderophore as cellular iron carriers is a new approach. In this research, for the first time the potential of siderophore in increasing the Fe enrichment of Saccharomyces boulardii (S. boulardii), which is important because of its probiotic properties and resistance to different stresses, has been investigated to produce of potential iron supplements. For this purpose, siderophore was produced by Pseudomonas aeruginosa (P. aeruginosa). Siderophore impact, along with ten other independent process variables, has been studied on the efficiency of iron biotransformation by the Plackett-Burman design (PBD). The results showed that the highest biotransformation yield was 17.77 mg Fe/g dry cell weight (DCW) in the highest biomass weight of 9 g/l. Iron concentration is the most important variable, with contributions of 46% and 70.79% for biomass weight and biotransformation, respectively, followed by fermentation time, agitation speed, and KH2PO4 concentration. But increasing the level of siderophore and zinc led to a significant negative effect. siderophore inefficiency may be attributed to the absence of membrane receptors for pyoverdine (Pvd) and pyochelin (Pch) siderophores. Also, the steric hindrance of the cell wall mannan, the stickiness and sediment ability of the yeast, can create limitations in the absorption of elements. Such yeast can be used as a potential source of iron even for vegetarians and vegans in the form of medicinal and fortified food products to improve the treatment of anemia. It is recommended that further research be focused on increasing the iron enrichment of yeast by overcoming the structural barrier of the cell wall, investigating factors affecting membrane permeability and iron transport potential of other types of siderophores.

Anti-oxidative activity of probiotics; focused on cardiovascular disease, cancer, aging, and obesity.

By disturbing the prooxidant-antioxidant balance in the cell, a condition called oxidative stress is created, causing severe damage to the nucleic acid, protein, and lipid of the host cell, and as a result, endangers the viability of the host cell. A relationship between oxidative stress and several different diseases such as cardiovascular diseases, cancer, and obesity has been reported. Therefore, maintaining this prooxidant-antioxidant balance is vital for the cell. Probiotics as one of the potent antioxidants have recently received attention. Many health-promoting and beneficial effects of probiotics are known, and it has been found that the consumption of certain strains of probiotics alone or in combination with food exerts antioxidant efficacy and reduces oxidative damage. Studies have reported that certain probiotic strains implement their antioxidant effects by producing metabolites and antioxidant enzymes, increasing the antioxidant capacity, and reducing host oxidant metabolites. Therefore, we aimed to review and summarize the latest anti-oxidative activity of probiotics and its efficacy in aging, cardiovascular diseases, cancer, and obesity.

Safety Evaluation and antioxidant potential of new probiotic strain Bacillus subtilis (NMCC-path-14) in Balb/c mice by sub-acute repeated dose toxicity.

Probiotics have recently gained significant interest for their possible therapeutic effects in treating numerous health conditions. Probiotics containing Bacillus subtilis have been shown to have several health benefits, most notably in preventing diarrhea and gastrointestinal problems. A novel probiotic strain, Bacillus subtilis (NMCC-path-14), isolated from the rumen of a Nilli Ravi Buffalo, was evaluated for 28-day repeated dose toxicity in Balb/c mice. The NMCC-path-14 in low dose (1 × 108 CFU/ml) and high dose (1 × 1010 CFU/ml) was administered to the mice through gavage regularly. After 28 days of treatment, it was discovered that the no-observed-adverse-effect level (NOAEL) for NMCC-path-14 wasgreater than 1 × 1010 CFU/animal/day. This study also revealed no treatment-related changes in clinical parameters, body weight, gross pathology, or histology. Food consumption, hemoglobin, hematocrit, red blood cell counts, and colon length increased, while total/differential leukocyte count and platelets remained unchanged. The administration of NMCC-path-14 also resulted in decreased bilirubin and creatinine levels. Furthermore, NMCC-path-14 also displayed a promising antioxidant potential by increasing the antioxidant enzymes (GST, GSH, and CAT) and decreasing oxidant enzyme (MDA and NO) levels in vital organs like the liver, kidneys, spleen, and colon. TheNMCC-path-14also decreased the pathogenic bacterial population while increasing the beneficial population. Given the lack of adverse effects observed after NMCC-path-14 treatment, this strain is safe and must be considered as a potential probiotic in humans.

Colonic long-term retention and colonization of probiotics by double-layer chitosan/tannic acid coating and microsphere embedding for treatment of ulcerative colitis and radiation enteritis.

Oral probiotics can alleviate enteric inflammations but their rapid transit through the gut limits their retention and colonization in the colon. Here, a novel strategy integrating the bacterial double-layer coating and hydrogel microsphere embedding techniques was used to highly enhance the colonic retention and colonization efficiency of Lactobacillus rhamnosus GG (LGG). LGG was coated by the double layers of chitosan (CS) and tannic acid (TA), and then embedded in calcium alginate (CA) hydrogel microspheres to form LGG@CT@CA. The microspheres resisted gastric liquids, improving LGG safe transit through the stomach to reach the colon. LGG@CT was rapidly released in the colon due to the good swelling of hydrogel microspheres. More importantly, LGG exhibited long-term retention up to 7 days in the colon and colonized the deep site of the colonic mucosa. LGG@CT@CA had a high therapeutic efficiency of ulcer colitis with the long colon and the low intestinal permeability of colonic tissues. LGG@CT@CA also alleviated the small intestinal damage induced by irradiation and the survival rates were improved. The mechanisms included local ROS decrease, IL-10 increase, and ferroptosis reduction in the small intestine. The oral colon-targeted system holds promise for oral probiotic therapy by the long-term retention and colonization in the colon.

The Efficacy and Safety of Probiotic Combinations Lobun Forte® Versus Renadyl® in Patients With Chronic Kidney Disease: A Comparative, Phase IV, Randomized, Open-Label, Active-Controlled, Parallel Study.

BACKGROUND: Chronic kidney disease (CKD) often leads to gut microbiota imbalance, accelerating disease progression and increasing uremic toxins and inflammation. We conducted a randomized clinical trial comparing outcomes between two multi-strain probiotic supplements Lobun Forte® (Sanzyme P Ltd, Hyderabad, India) containing Streptococcus thermophilus, Lactobacillus acidophilus, Bifidobacterium longum, and Bacillus coagulans and Renadyl® (Kibow Biotech, LLC., Pennsylvania, United States) containing S. thermophilus, L. acidophilus, and B. longum. MATERIALS AND METHODS: Sixty patients with stage 3-4 CKD were randomized to receive either Lobun Forte (n=30) or Renadyl (n=30) for six months, with each supplement providing 45 billion CFU/capsule, twice daily. Primary outcomes included quality of life (QoL) (Short-Form 8 (SF-8) score), reductions in uremic toxins (p-cresyl sulfate (PCS), 3-indoxyl sulfate (IS), indole-3-acetic acid (IAA)), blood urea nitrogen (BUN), serum creatinine, and serum uric acid. Secondary outcomes assessed oxidative stress, inflammatory biomarkers, and estimated glomerular filtration rate (eGFR). Results: Both Lobun Forte and Renadyl groups showed significant improvements in QoL, with Lobun Forte achieving a 53.5% improvement (16.43 point increase) and Renadyl a 51.1% improvement (15.27 point increase) in SF-8 scores (p < 0.0001). The levels of IS decreased significantly in both groups (p < 0.0001), with Lobun Forte reducing IS by 29.72% and Renadyl by 24.20%. In terms of other uremic toxins, Lobun Forte showed non-significant (p > 0.05) reductions in mean PCS (7.63%) and IAA (15.57%), whereas Renadyl demonstrated a significant (p = 0.0314) decrease in PCS (20.75%) and a non-significant (p > 0.05) reduction in IAA (12.35%). Both groups showed significant (p < 0.0001) reductions in BUN and serum creatinine levels. Serum uric acid levels showed a significant (p = 0.0448) reduction with Lobun Forte while Renadyl exhibited a non-significant reduction (p = 0.1034). Lobun Forte significantly (p = 0.0359) reduced mean high-sensitivity C-reactive protein (hsCRP) levels, while Renadyl showed a non-significant reduction (p = 0.0876). Both groups had non-significant reductions in interleukin-6 and tumor necrosis factor-alpha levels (p > 0.05). Further, both groups experienced significant (p < 0.0001) increases in mean glutathione levels and nitric oxide levels. Additionally, Renadyl resulted in a significant reduction in mean malondialdehyde, whereas Lobun Forte showed a non-significant reduction. Both probiotics significantly (p < 0.0001) improved eGFR, with Lobun Forte increasing it by 40.4% and Renadyl by 36.9%. Both probiotics were well tolerated, with a favorable safety profile throughout the study. Conclusion: Both Lobun Forte and Renadyl effectively improve the quality of life in patients with stage 3-4 CKD by modulation of uremic toxins, renal parameters, inflammatory biomarkers, oxidative biomarkers, and eGFR. These findings suggest that both probiotics may help delay CKD progression by modulating the gut-kidney axis.

Probiotaceuticals: Back to the future?

Probiotic research has undergone some exciting and unanticipated changes in direction since the 2010 commentary by GSH, which speculated on probiotics being ultimately utilized as "factories" capable of releasing pharmaceutical-grade metabolites with therapeutic potential for a wide range of primarily gastrointestinal disorders. Indeed, the unrelenting search for new alternatives to antibiotics has further stimulated the development of "next-generation" probiotics. Postbiotics, defined as inanimate microorganisms and/or their components that confer a health benefit on the host, remain at the forefront of current probiotic research, with increasing numbers of probiotic species, strains, and substrains now being identified and further exploited as pharmabiotics; probiotics with a proven pharmacologic role in health and disease that have been subjected to clinical trial prior to approval by regulatory bodies. However, perhaps the most unanticipated probiotic development over the past 15 y has been the emergence of psychobiotics with the potential to improve aspects of mental health, such as depression and anxiety, through the release of bioactive metabolites. Moreover, the recent identification of pharmacobiotics, probiotics capable of facilitating the effectiveness of conventional pharmaceutical drugs, is opening new avenues for probiotic applications to combat a range of diseases, including cancers of the digestive system. Although in its infancy, recent reports of oncobiotics with antineoplastic properties are further expanding the potential for certain next-generation probiotics to impact current cancer treatment regimens and possibly even contribute to cancer prevention. Looking to the next 15 y of probiotic development, one could perhaps predict the ultimate development of regulatory-approved xenopostbiotic formulations comprising metabolites with the capacity to improve digestive health, decrease the severity of intestinal disease, and increase the effectiveness of conventional pharmaceuticals, whereas simultaneously improving cognitive functioning and mental welfare. Although speculative, these xenopostbiotic formulations could prove especially effective for the adjunctive treatment of serious chronic diseases such as cancer.

Antioxidant Peptides Derived from Cheese Products via Single and Mixed Lactobacillus Strain Fermentation.

Probiotics are used in cheese fermentation to endow the product with unique functional properties, such as enhanced flavor and aroma development through proteolysis and lipolysis. In this study, two probiotic Lactobacillus strains, Lactobacillus plantarum A3 and Lactobacillus reuteri WQY-1, were selected to develop new probiotic cheeses in the form of single- and mixed-strain starters. The results demonstrated that the L. plantarum A3 single-strain group and the L. plantarum A3/L. reuteri WQY-1 mixed fermentation group exhibited superior product performance, particularly the release of functional hydrolysates during cheese ripening. Furthermore, Label-free quantitative proteomic analysis revealed 26 unique antioxidant peptides in the L. plantarum A3 single-strain group and 53 in the L. plantarum A3/L. reuteri WQY-1 mixed fermentation group. Among these, CMENSAEPEQSLACQCL (ß-lactoglobulin), CMENSAEPEQSLVCQCL (ß-lactoglobulin), and IQYVLSR (κ-casein) have been found to possess potential antioxidant properties both in vitro and in vivo. This confirmed that milk-derived protein peptides in cheese products exhibit potential antioxidant functions through the hydrolysis of probiotic strains.

Colon Cancer Cells Treated with Lacticaseibacillus casei Undergo Apoptosis and Release DAMPs Indicative of Immunogenic Cell Death.

Probiotic bacteria, and especially lactic acid bacteria, have long been known to wield a variety of health-beneficial effects, including antioxidant, antimicrobial, anti-inflammatory, immunomodulatory, and anticancer activities. However, our understanding of the mechanisms involved in these activities remains incomplete. In this study, we wished to investigate the processes that give rise to the anticancer activity of Lacticaseibacillus casei ATCC393 and the possibility that immunogenic cell death of cancer cells can be induced following treatment with this probiotic. In both cell lines that we have examined, we detected notable pro-apoptotic signaling, including the upregulation of death receptors, that culminated in the activation of caspase 3, the endpoint and most characteristic effector molecule of all pro-apoptotic cascades. In addition, we identified damage-associated molecular patterns associated with immunogenic cell death. Calreticulin exposure on the outer cell membrane, HMGB1 translocation outside the nucleus and depletion of intracellular ATP was evident in both cancer cell lines treated with the probiotic, while expression of type I interferons was upregulated in CT26 cells. Our findings suggest that treatment with the probiotic induced apoptosis in cancer cells, mediated by extrinsic death receptor signaling. Moreover, it resulted in the release of molecular signals related with immunogenic cell death and induction of cancer cell-specific adaptive immune responses.

Effects of cholesterol-lowering probiotic fermentation on the active components and in vitro hypolipidemic activity of sea buckthorn juice.

Sea buckthorn has lipid-lowering properties and is widely used in the development of functional foods. In this study, a probiotic (Lactobacillus plantarum, Lp10211) with cholesterol-lowering potential and acid and bile salt resistant was screened for the fermentation of sea buckthorn juice. Changes in the active ingredients, such as sugars and phenolics, before and after fermentation, as well as their in vitro lipid-lowering activities, were compared. The contents of reducing and total sugars decreased substantially after fermentation. Lp10211 primarily utilized fructose for growth and reproduction, with a utilization rate of 76.9%. The phenolic compound content of sea buckthorn juice increased by 37.06% after fermentation and protected the phenolic components from degradation (protocatechuic and p-coumaric acids) and produced new polyphenol (shikimic acid). Enhanced inhibition of pancreatic lipase activity (95.42%) and cholesterol micellar solubility (59.15%) was evident. The antioxidant properties of the fermentation broth were improved. Notably, Lp10211 preserved the color and reversed browning in sea buckthorn juice. The collective findings indicate that fermentation of sea buckthorn juice by Lp10211 may enhance the functional components and lipid-lowering activity of sea buckthorn, which may provide a new approach for the development of lipid-lowering foods.

Intestinal probiotic-based nanoparticles for cytotoxic siRNA delivery in immunotherapy against cancer.

Immunogene therapy has emerged as strategy against cancer by introducing immune-stimulating components into gene therapy. However, there is still a need for an ideal platform to achieve both immune stimulation and efficient gene delivery. Lactobacillus reuteri has potential immunomodulatory activity owing to its unique antigenicity, which is potentially relevant to cancer progression. Here, we designed a novel non-viral siRNA vector (DMPLAC) by encapsulating Lactobacillus reuteri lysate in DMP. DMPLAC can promote maturation and activation of immune cells, increase infiltration of APC and cytotoxic T cells in tumor microenvironment, and exhibit tumor suppressive effects. Loading of siRNA targeting Stat3, DMPLAC/siStat3 further inhibits tumor in multiple models. We designed a strategy that combines immune activation with Stat3 silencing, triggering an immune response and tumor killing. This dual-functional design provides a new choice in development of effective immunogene therapy.

Effects of derivatization and probiotic transformation on the antioxidative activity of fruit polyphenols.

Fruits contain numerous polyphenols in the form of conjugates, which exhibit low antioxidant activity. Probiotic fermentation is a strategy to improve the antioxidant activity of these conjugated polyphenols by modifying their structure. However, the mechanisms underlying the effects of functional groups and derivatizations on the antioxidative activities of polyphenols and the antioxidation enhancement by probiotic biotransformation haven't been comprehensively explored. This review aimed to explore the structure-antioxidant activity relationships of four functional groups and three derivatizations in flavonoids and phenolic acids. Further, the review elucidated the antioxidant mechanisms underlying the biotransformation of flavonoids and phenolic acids as glycoside, methylated, and ester conjugates by probiotic biotransformation. Deglycosylation, demethylation, and hydrolysis catalyzed by enzymes produced by Bifidobacterium and Lactobacillus facilitated the conversion of conjugated polyphenols into flavonoids and phenolic acids with hydrolyzed forms and highly active functional groups, thereby increasing hydrogen supply and electron transfer capacity to enhance the antioxidant activity.

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Bariatric surgery may cause intestinal microecological environment imbalance due to changes in gastrointestinal anatomy. Some patients may have complications, even regain weight. Probiotics can act on intestinal mucosa, epithelium and gut-associated lymphoid tissue to improve the intestinal microecological environment of obese patients after bariatric surgery. Probiotics can promote the production of intestinal antibacterial substances, bind specifically to receptors, decrease intestinal pH value, reduce the inflammatory factors, thus helping patients lose weight and lower blood sugar levels after bariatric surgery. Probiotics can produce lactic acid, acetic acid, lactase, etc., inhibit the growth of harmful bacteria, improve gastrointestinal symptoms of patients after bariatric surgery. Probiotics can activate the AMP-activated protein kinase signaling pathway, improve lipid metabolism, and promote the recovery of symptom indicators of nonalcoholic fatty liver disease after bariatric surgery. Probiotics can regulate the release of neurotransmitters or metabolites by the microbiota through the gut-brain axis to affect brain activity and behavior, thus helping patients improve bad mood after bariatric surgery. This article describes the intestinal microecological environment of obese patients and the change mechanism after bariatric surgery and summarizes the effects and possible mechanisms of probiotics in improving the intestinal microecological environment of obese patients after bariatric surgery, in order to provide references for promoting the clinical application of probiotics.

Physicochemical, Sensory, and Microbiological Analysis of Fermented Drinks Made from White Kidney Bean Extract and Cow's Milk Blends during Refrigerated Storage.

Due to its low dietary impact and bioactive compounds, such as polyphenols and flavonoids, white kidney bean extract is an attractive raw material for fermented drinks. It can be utilized either on its own or blended with cow's milk, offering a promising solution to help meet dairy product demand during mid-season shortages. Therefore, this study aimed to explore the physicochemical characteristics, sensory properties, and microbiological profile of fermented milk-like drinks made from white kidney bean extract, cow's milk and their blends during 28 days of storage at 4 °C. Three blends of fermented milk-like drinks (FMLDs) were prepared from different ratios of cow's milk (CM) and kidney bean extract (BE): FMLD1 (CM 30%:BE 70%); FMLD2 (CM 50%:BE 50%), FMLD3 (CM 70%:BE 30%), along with plain fermented kidney been extract (FBE; CM 0%:BE 100%), and plain fermented cow's milk (FCM; CM 100%:BE 0%). The mixtures were pasteurized at 92 °C for 25 min and fermented with a probiotic-type starter culture (S. thermophilus, B. bifidum, L. acidophilus) at 43 °C. FBE exhibited the lowest levels of carbohydrates (2.14%), fat (0.11%), and protein (1.45%) compared to fermented cow's milk and blends. The FBE and the fermented blends with a higher ratio of bean extract had lower viscosity and lactic acid contents, greener hue, more pronounced aftertaste and off-flavors, and received lower overall acceptability scores. Although the FCM had higher counts of S. thermophilus and L. acidophilus, the FBE displayed significantly higher counts of B. bifidum. This study demonstrated the potential of using white kidney bean extract and its blends with cow's milk to create unique fermented products with a lower dietary impact, highlighting the importance of further optimizing the formulations to enhance sensory qualities and reduce the beany off-flavors in the products with added kidney bean extract.

Assessment of the Probiotic Properties of Yarrowia lipolytica Isolated from Cold-Pressed Olive Oil.

This research aimed to identify the probiotic features of Yarrowia lipolytica strains isolated from olive oils in Turkey. The in vitro survival capabilities of Y. lipolytica strains in gastric and pancreatic solutions were assessed. The hydrophobicity of Y. lipolytica strains was determined to be between 25.8% and 46.08% for xylene, 22.5% and 45.85% for chloroform, and 14.83% and 37.09% for ethyl acetate. In addition, auto-aggregation values were measured as 11.07-60.35%; 16.28-67.70% and 42.89-85.21% after 2, 4 and 24 h of incubation, respectively. The Y. lipolytica strains tested in this study demonstrated aggregation ability against the pathogens Escherichia coli ATCC 25922, Salmonella typhimurium ATCC 14028, Staphylococcus aureus ATCC 25923 and Listeria monocytogenes ATCC 7644. Antibiotic resistance and hemolytic activities were also checked to ensure the safety of the Y. lipolytica. Cholesterol removal by Y. lipolytica strains ranged from 12.30% to 47.42%, and their free radical scavenging activity varied between 2.85% and 39.10%. Out of 13 Y. lipolytica samples from 10 different olive oil sources, Y. lipolytica Y6, Y7, and Y11 exhibited the best strains with probiotic potential properties. This study discovered that Y. lipolytica with probiotic properties can be isolated in olive oil samples, a finding that has not been previously documented in the literature and may have potential industrial applications.

Antitumor properties of traditional lactic acid bacteria: Short-chain fatty acid production and interleukin 12 induction.

This paper presents an in vitro evaluation of antitumor properties through producing short-chain fatty acids and inducing interleukin 12. In addition, it offers the most important and functional probiotic properties of 24 Lactobacillus gasseri, Lactiplantibacillus plantarum, Lactobacillus acidophilus, and Limosilactobacillus fermentum strains isolated from humans, foods, and fermented foods. To this end, survival in an acidic environment (pH = 2.5), tolerance in bile salt, viability in the presence of pepsin-pancreatin, adhesion percentage, antibiotic resistance, auto-aggregation, and potential percentage of co-aggregation are studied in contact with three human intestinal pathogens. These pathogens are Escherichia coli O157: H7 NCTC 12900, Salmonella enterica subsp. enterica ATCC 13076, and Listeria monocytogenes ATTC 7644. Also, in vitro induction amount of IL-12 in mouse splenocytes is investigated to evaluate antitumor properties by 19 strains of L. gasseri and L. plantarum along with the development of short-chain fatty acids (SCFA) by 5 strains of L. fermentum and L. acidophilus. Gas Chromatography Flame Ionization Detector (GC-FID) and enzyme-linked immunosorbent assay (ELISA) were used to measure short-chain fatty acids and IL-12, respectively. All strains had high viability under acidic conditions. The highest levels of pancreatin and pepsin resistance were found in strains LF56, LF57, LF55, OF, and F and strains LF56, LF57, and A7, respectively. All strains except LF56 had high resistance to bile salts. L. gasseri 54C had the highest average adhesion score (hydrophobicity) of 62.9 % among 19 strains. Despite the susceptibility of different strains of L. plantarum to the tested antibiotics, M8 and M11, S2G, A7, LF55, LF57, and 5G were resistant to kanamycin and chloramphenicol, respectively. Also, 21G was resistant to ampicillin, LF56 to tetracycline and M8, and M11, LF56, and 21G to Erythromycin. In addition, L. gasseri showed moderate resistance to ampicillin, erythromycin, and tetracycline, while L. fermentum ATCC 9338 showed good resistance to ampicillin, erythromycin, and chloramphenicol. In this respect, L. plantarum LF56 and gasseri 54C had the highest average auto-aggregation and co-aggregation against three pathogenic bacteria, respectively. The highest and lowest levels of acetic acid as short-chain fatty acids were produced by L. fermentum 19SH isolated from Horre 41.62 and L. fermentum 21SH from fermented seeds 27.047, respectively. Moreover, L. fermentum, with the OF code of traditional-fermented food origin, produced the most isobutyric acid, butyric acid, and valeric acid, with values of 0.6828, 0.74165, and 0.49915 mmol, respectively. L. fermentum isolated from the human origin with code F produced the most isovaleric acid of 1.1874 mmol. All the tested strains produced good propionic acid except L. fermentum 21SH from fermented seeds. Among strains, L. plantarum M11 isolated from milk and L. gasseri 52B from humans had the highest in vitro induction of IL-12, which is probably related to their cell wall compositions and structure.

Different polysaccharide-enhanced probiotic and polyphenol dual-functional factor co-encapsulated microcapsules demonstrate acute colitis alleviation efficacy and food fortification.

Probiotics and polyphenols have multiple bioactivities, and developing co-encapsulated microcapsules (CM) is a novel strategy to enhance their nutritional diversity. However, the development of CMs is challenged by complicated processing, single types, and unclear in vivo effects and applications. In this study, the co-microencapsulations of polyphenol and probiotic were constructed using pectin, alginate (WGCA@LK), and Fu brick tea polysaccharides (WGCF@LK), respectively, with chitosan-whey isolate proteins by layer-by-layer coacervation reaction, and their protective effects, in vivo effectiveness, and application potential were evaluated. WGCA@LK improved the encapsulation rate of polyphenols (42.41 %), and remained high viability of probiotics after passing through gastric acidic environment (8.79 ± 0.04 log CFU/g) and storage for 4 weeks (4.59 ± 0.06 log CFU/g). WGCF@LK exhibited the highest total antioxidant activity (19.40 ± 0.25 µmol/mL) and its prebiotic activity removed the restriction on probiotic growth. WGCA@LK showed strong in vitro colonic adhesion, but WGCF@LK promoted in vivo retention of probiotics at 48 h. WGCF@LK showed excellent anti-inflammatory effects and alleviated symptoms of acute colitis in mice. These findings provide unique insights into the fortification of probiotic-polyphenol CMs by different polysaccharides and the development of novel health foods with rich functional hierarchies and superior therapeutic effects.

Integrating fruit by-products and whey for the design of folate-bioenriched innovative fermented beverages safe for human consumption.

Global concerns over folate deficiency, the risks of excessive synthetic folic acid consumption, and food loss implications for environmental sustainability and food security drive needs of innovative approaches that align food by-product valorisation with folate bio-enrichment. This study explored the use of three fruit by-products extracts (grape, passion fruit, and pitaya) and whey to develop a folate bio-enriched fermented whey-based beverage. Three strains (Lacticaseibacillus rhamnosus LGG, Bifidobacterium infantis BB-02, and Streptococcus thermophilus TH-4) were tested for folate production in different fermentation conditions in modified MRS medium and in a whey-based matrix prepared with water extracts of these fruit by-products. B. infantis BB-02 and S. thermophilus TH-4, alone and in co-culture, were the best folate producers. The selection of cultivation conditions, including the presence of different substrates and pH, with grape by-product water extract demonstrating the most substantial effect on folate production among the tested extracts, was crucial for successfully producing a biofortified fermented whey-based beverage (FWBB). The resulting FWBB provided 40.7 µg of folate per 100 mL after 24 h of fermentation at 37 °C, effectively leveraging food by-products. Moreover, the beverage showed no cytotoxicity in mouse fibroblast cells tests. This study highlights the potential for valorising fruit by-products and whey for the design of novel bioenriched foods, promoting health benefits and contributing to reduced environmental impact from improper disposal.

Improving Vaccine Response through Probiotics and Micronutrient Supplementation: Evaluating the Role of TLR5 in Adult Female BALB/c Mice.

BACKGROUND: The role of probiotics and micronutrients in improving immune system function and response to vaccination has been proven. Hence, this study aimed to investigate the effects of probiotics enriched with micronutrients on the immunogenicity of PastoCovac® vaccine. METHODS: The probiotic supplement BioBoost® and PastoCovac® vaccine, which contain six expressed receptor-binding domains (RBD) and conjugated with tetanus toxin, were administered concurrently. The safety and efficacy were assessed by determining Immunoglobulin G (IgG) antibody titers to RBD and cytokines, mRNA expression of Toll-like Receptors (TLRs) 5, and clinical symptoms. RESULTS: Results revealed that the administration of the probiotics enriched with micronutrients and vitamins for 14 days before the first vaccine dose, followed by continued supplementation for 14 days after the first dose, and in conjunction with the second vaccine dose, yielded the most significant elevation in interleukin 4 (IL-4), Tumor Necrosis Factor-alpha (TNF alpha), Interferon-gamma (IFN-gamma), and anti-SARS-CoV-2 RBD IgG levels within the supernatant samples collected from spleen cultures with the highest expression of TLR5 genes in intestinal samples, compared to the control group. CONCLUSION: Our results indicated that the inclusion of probiotics enriched with micronutrients and vitamins significantly enhanced the immunogenicity of the PastoCovac® vaccine. Based on the recommendation to administer third and fourth vaccine doses, particularly for vulnerable and elderly individuals, the utilization of supplements containing probiotics is expected to favorably influence immune responses.

Impact of a Functional Dairy Powder and Its Primary Component on the Growth of Pathogenic and Probiotic Gut Bacteria and Human Coronavirus 229E.

Milk boasts an array of potent bioactive compounds, such as lactoferrin (Lf), immunoglobulins, and functional proteins, all delivering substantial therapeutic benefits. In this study, Immune Powder (a functional dairy formulation) and its primary component called Fractionated Milk Protein (FMP) containing Lf, zinc, and immunoglobulins and formulated by Ausnutria Pty Ltd. were evaluated for their potential broad-spectrum pharmacological activity. In particular, this study investigated the antibacterial (against pathogenic Escherichia coli), prebiotic (promoting Lactobacillus delbrueckii growth), anti-inflammatory (inhibition of NO production in RAW264.7 macrophages), and antiviral (against human coronavirus 229E) effects of the samples. In addition, the impact of simulated gastric digestion on the efficacy of the samples was explored. LCMS-based proteomics was implemented to unveil cellular and molecular mechanisms underlying antiviral activity. The Immune Powder demonstrated antibacterial activity against E. coli (up to 99.74 ± 11.47% inhibition), coupled with prebiotic action (10.84 ± 2.2 viability fold-change), albeit these activities diminished post-digestion (p < 0.01). The Immune Powder effectively mitigated NO production in lipopolysaccharide-stimulated RAW264.7 macrophages, with declining efficacy post-digestion (p < 0.0001). The Immune Powder showed similar antiviral activity before and after digestion (p > 0.05) with up to 3-fold improvement. Likewise, FMP exhibited antibacterial potency pre-digestion at high concentrations (95.56 ± 1.23% inhibition at 125 mg/mL) and post-digestion at lower doses (61.82 ± 5.58% inhibition at 3906.25 µg/mL). FMP also showed enhanced prebiotic activity post-digestion (p < 0.0001), NO inhibition pre-digestion, and significant antiviral activity. The proteomics study suggested that the formulation and its primary component shared similar antiviral mechanisms by inhibiting scavenger receptor binding and extracellular matrix interaction.

Second-generation snacks prepared from quinoa with probiotic. Physicochemical properties, in vitro digestibility, antioxidant activity and consumer acceptability.

Food security is a pressing issue, emphasizing the necessity for food designs that address the current geopolitical and geoeconomic challenges. This study evaluates the impact of including different percentages (10 %, 20 %, 30 %, and 60 %) of high protein quinoa flour (HPQF) in the development of a new rice-based snack. The aim is to create four snack formulations with a protein content enriched with probiotics, surpassing those currently available in the market. Probiotics Bacillus coagulans were added at a 0.1 % concentration. Once the rice flour and quinoa flour are mixed, they are mixed with the probiotic, to move on to the extrusion process. Following the incorporation of probiotics, the snacks were packaged in a modified atmosphere, and their physicochemical properties, Bacillus coagulans probiotic viability, tolerance to artificial gastroenteric juice (TAGJ), starch digestibility, and sensory acceptance were assessed. Significant differences were observed in the expansion index, with the 60 % inclusion snack exhibiting the least expansion. Despite having a higher density, this snack reached a porosity index similar to that of the 20 % HPQF snack. Achieving a 17 % protein content in the snacks was possible with a 60 % inclusion rate. Texture was notably affected by the inclusion of HPQF, with snacks having higher inclusion levels showing increased hardness. Probiotic viability evaluation consistently remained above 106 UFC/g of snack, while TAGJ exhibited a viability of 75 %. Although HPQF inclusion led to a decrease in the glycemic index (GI), snacks still maintained a GI above 70 %. Regarding antioxidant properties, snacks with 60 % HPQF inclusion displayed superior results, reaching 35.29, 5.52, and 13.74 µmol of AA/g, measured via ABTS, DPPH, and FRAP methods, respectively. These findings demonstrate a heightened antioxidant capacity compared to other formulations. Our results indicate that the new probiotic snack serves as a rich source of protein and probiotics and is well-received sensorially. However, it is worth noting that it falls within the category of high GI foods, prompting the need for future studies aimed at reducing this parameter.