Probiotic Cells Research Articles

Grape pomace high-methoxyl pectin: A new prebiotic stabilizer for low-fat synbiotic yogurt gels - Optimization and characterization.

High-methoxyl pectin (HMP, 72.5 % esterification degree and galacturonic acid content of 67.9 %) was extracted from grape pomace using a sequential ultrasound-microwave extraction. The extracted HMP was used to develop low-fat synbiotic set yogurts containing probiotic cells. Higher grape pomace pectin (GPP) concentrations (0.5-2 %) increased the probiotic bacterial population of Lactobacillus acidophilus LA-5 and Bifidobacterium bifidum BB-12. Higher cell viability was observed for L. acidophilus LA-5 compared to B. bifidum BB-12. A response surface optimization showed that the presence of 8.08 Log CFU mL-1L. acidophilus LA-5 and 1.88 % HMP experimentally resulted in the best probiotic viability (10.83 ± 0.11 Log CFU mL-1), overall acceptability (8.03 ± 0.06), and pH (4.25 ± 0.05) values. Compared to pectin-free probiotic yogurts, the optimal yogurt gels presented higher probiotic survivability, lower syneresis, and superior storage-dependent sensory attributes during 21 days of storage. However, a 14-day storage period was generally deemed suitable. The GPP-containing yogurt compared to the pectin-free sample exhibited higher colloidal stability with a larger particle size (433.8 nm vs. 272.5 nm) and lower zeta potential (-20.4 mV vs. -10.6 mV). Field emission-scanning electron (FE-SEM) and fluorescent (FLM) microscopy images confirmed a denser microstructure for GPP-enriched yogurts. The chemical interactions in the yogurt were not affected by enriching with GPP as investigated by FTIR, whereas the steady and dynamic rheological properties were significantly improved. GPP-enriched yogurt had a firmer gel structure with a larger linear region and lower G' compared to the control, indicating a semi-solid state. The GPP as a multi-functional prebiotic ingredient would be promising in designing healthier food products.

The development of microencapsulated lactic acid bacteria and its application on yoghurt powder products

The probiotics were susceptible to damage in commercial starters due to improper environmental conditions. In this study, the encapsulation technique was applied to improve probiotics survival using alginate, inulin (commercial and beneng taro-source), skim milk and their combination; which is prepared by the extrusion method. The survival of encapsulated probiotics in adverse environmental conditions was investigated by exposing them to low pH and heat stress. The encapsulation yield and diameter of the encapsulated probiotics were also measured before their application to yoghurt powder. Both heat stress and pH 2 resulted in the decrease of the number of viable free cells and viable encapsulated cells by about 5 log cycles and 3 log cycles, respectively. The encapsulation yield was >98% in all treatments and the diameter of probiotics was increased significantly around >2.5 mm by the encapsulation method. The viability of the encapsulated probiotic cell was decreased 1 log cycle after applying in powdered yoghurt while without encapsulation the cell count was much lower. Thus, the microencapsulation method has proven as an effective protection technique against adverse environments, especially if applied as a powder product.

A novel immunopharmacological biocompound

The publication is devoted to topical issues of experimental study of new biocompounds, based on the creation of a consortium of biological composite compounds – biological active substances (metabiotics) produced by strains 59T and 60T of saprophytic compounds of the genus Bacillus subtilis. These strains are very promising for the creation of hepatopicts, new medicinal substances, in the creation and development of a new pharmacological class of hepatoprotectors. Previous studies have shown the safety and hepatoprotective effect of biologically active drugs. It is worth noting that the very effective compounds are the produced biologically active substances – metabolites of bacterial origin, on the basis of which it seems appropriate to create a new class of drugs – metabiotics. The absence of vegetative probiotic cells in such compounds will reduce the additional immune load on the body. The combined use of these compounds provides a potentiated pharmacological effect. In this regard, it seemed appropriate, under the conditions of modeling toxic liver damage by carbon tetrachloride, to conduct an experimental assessment of the immunotropic effect of biologically active substances (BAS) on laboratory animals in order to confirm the effect on cellular factors of immunity. The purpose of the study was to study the effect of the combined use of dietary supplements produced by Bacillus subtilis microorganisms on indicators of cellular immunity in laboratory animals with toxic liver damage. Acute toxic hepatitis was reproduced in white laboratory rats with repeated intragastric administration of carbon tetrachloride. The comparison drug was a registered hepatoprotective drug – ursosan. The immunotropic effect was assessed using factors such as: phagocytic activity of macrophages and neutrophils (FA), NBT test, quantitative assessment of antibody-forming cells, T and B lymphocytes. As a result of the studies performed, reliable data were obtained on an increase in the number of T and B lymphocytes, antibody-forming cells, as well as an increase in FA, which confirms the activation of all parts of cellular immunity in conditions of acute toxic liver damage. The data obtained allow us to recommend the studied biocompound for the creation of new drug candidates of microbiological origin, hepatoprotective agents with immunotropic effects.

Formulation of synbiotic buttermilk powder with higher viability of probiotic cells

The present study focused on the use of trehalose (TRE), whey protein concentrate (WPC), pullulan (PUL), and calcium lactate gluconate (CAL) as a protectant in the formulation of synbiotic buttermilk powder by application of spray drying. The spray drying of the solution mixture was carried out at an inlet temperature of 150 °C, a feed flow rate of 15mL/min, and compressed air at 3 kg/cm2 while keeping the outlet temperature constant at 70 ± 5 °C. TRE4 + CAL0.5 showed good results with the viability of Bifidobacterium bifidum as 6.04 ± 0.13 after 16 weeks followed by TRE2 + CAL0.5, TRE4, CAL0.5, WPC6, PUL4, and Control. The addition of protectants and Fructooligosaccharide (FOS) as a prebiotic did not affect the powder properties. The moisture content, water activity, bulk density, hygroscopicity, solubility, particle size, and scanning electron microscopy (SEM) of SBMP were better than the control sample for 17 weeks. SEM results showed spherical shapes without any cracks for SBMP containing TRE4 + CAL0.5. The present work demonstrated that the formulation containing TRE4 + CAL0.5 showed excellent results in terms of the viability of probiotic cells and also physicochemical properties of powder during storage for 16 weeks at 4 ± 1 °C.

Ameliorated Viability of Lactic Acid Bacteria in Fruit Juice Isolated from Indigenous Dahi with Prebiotics (Asparagus falcatus and Zingiber officinale)

Dahi is a fermented milk product containing probiotic lactic acid bacteria. This study aimed to isolate, identify, and characterize lactic acid-producing bacteria from native Dahi and evaluate their viability in orange juice using natural prebiotics. Dahi samples were obtained from local shops in Chattogram and Bogura, Bangladesh. Lactic acid-producing bacteria were isolated using MRS (de Mann Rogosa and Sharpe) medium. The isolated bacteria were identified through colony morphology, biochemical tests, and probiotic characteristics. Molecular identification was performed using polymerase chain reaction (PCR) targeting conserved 16S rDNA regions. Isolates of the genus Lactobacillus and Lactococcus lactis sp. Lactis were confirmed and used to develop probiotic orange juice. Prebiotics (Asparagus falcatus and Zingiber officinale) were added to the juice to support probiotic growth. The inoculated cell’s viability and the juice’s physicochemical parameters were evaluated during fermentation (48 hours) and storage (28 days). All fruit juice samples showed a mean number of viable cells of at least 1×105 CFU/mL during the 48-hour fermentation and 28-day storage in the refrigerator. Using natural prebiotics positively affected the survival of lactic acid bacteria, as demonstrated by bacterial colony growth on Petri dishes. Developing probiotic fruit juice enriched with prebiotics could be an effective alternative for individuals allergic or intolerant to milk-based products. Incorporating lactic acid bacteria from native Dahi into orange juice, combined with natural prebiotics, resulted in viable probiotic cells throughout fermentation and storage.

Novel Encapsulation Approaches in the Functional Food Industry: With a Focus on Probiotic Cells and Bioactive Compounds.

Bioactive substances can enhance host health by modulating biological reactions, but their absorption and utilization by the body are crucial for positive effects. Encapsulation of probiotics is rapidly advancing in food science, with new approaches such as 3D printing, spray-drying, microfluidics, and cryomilling. Co-encapsulation with bioactives presents a cost-effective and successful approach to delivering probiotic components to specific colon areas, improving viability and bioactivity. However, the exact method by which bioactive chemicals enhance probiotic survivability remains uncertain. Co-crystallization as an emerging encapsulation method improves the physical characteristics of active components. It transforms the structure of sucrose into uneven agglomerated crystals, creating a porous network to protect active ingredients. Likewise, electrohydrodynamic techniques are used to generate fibers with diverse properties, protecting bioactive compounds from harsh circumstances at ambient temperature. Electrohydrodynamic procedures are highly adaptable, uncomplicated, and easily expandable, resulting in enhanced product quality and functionality across various food domains. Furthermore, food byproducts offer nutritional benefits and technical potential, aligning with circular economy principles to minimize environmental impact and promote economic growth. Hence, industrialized nations can capitalize on the growing demand for functional foods by incorporating these developments into their traditional cuisine and partnering with businesses to enhance manufacturing and production processes.

Antioxidant Activity and Oxidative Stress Survival of Limosilactobacillus reuteri LR92 in Fermented Milk with Juçara Pulp

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.

Fermented plant-based beverage supplemented with uvaia (Eugenia pyriformis) pulp: an innovative and pioneering approach to diversify plant-based diet product market

Over the years, there has been an increase in demand for plant-based foods as alternatives. In line with this, this work explores the production and in vitro digestion of a fermented plant-based beverage (FPBB) produced with pea and rice proteins and 0% (FPBB-C), 5% (FPBB-5), and 10% (FPBB-10) uvaia pulp through lactic fermentation with Lacticaseibacillus rhamnosus GG. The in vitro gastrointestinal digestion process was conducted to assess the bioaccessibility of L. rhamnosus GG, total phenolic content (TPC), and antioxidant activity before and after simulating the gastrointestinal conditions. After 48 h of digestion, highly viable L. rhamnosus GG cells remained throughout the gastrointestinal system. FPBB-C (106.89%) and FPBB-5 (109.38%) exhibited higher survival rates than FPBB-10 (102.20%), indicating that these beverages have a higher prebiotic action potential. Compared with the non-digested samples, after 48 h of digestion, all samples exhibited a significant increase in TPC. The same behavior occurs for the antioxidant activity of FPBB-C, FPBB-5, and FPBB-10 by DPPH (4.06, 3.96, and 8.44 mg TEAC mL−1), ABTS (10.28, 11.06, 11.97 mg TEAC mL−1), and FRAP method (917.02, 863.87, and 1983.23 mg TEAC mL−1). Thirteen compounds were identified and quantified in uvaia pulp by HPLC-DAD-ESI-MS, particularly epigallocatechin gallate, quercetin-3-rhamnose, and quercetin-3-glucoside. Isorhamnetin was the main phenolic compound detected in the colon, assumably due to the conversion of quercetin-3-glucoside by the probiotic cells. In conclusion, as all counts were above 9 log CFU g−1, the FPBB formulations containing pea, rice protein, and uvaia pulp become a promising vehicle for carrying L. rhamnosus GG.

Porous starch-probiotics encapsulation for 3D-printed chocolate

This study aims to utilize 3D printing technology to prepare functional chocolates with high content and viability of probiotics. Firstly, in order to protect probiotics (L. plantarum, lpl) avoiding inactive by melting chocolate syrup, we used raw starches as different amylose/amylopectin ratios (up to ∼30% amylose) for preparing novel porous carriers to encapsulate them. Then the formed pores of porous starches (HPPS1:0,3:1,1:1,1:3,0:1) were identified by SEM and BET, and their ability to encapsulate lpl were compared (with HPPS3:1 showing the best behavior) based on the survival rate and gastrointestinal simulation system. Then HPPS3:1-lpl capsule was applied in 3D printed chocolate, with optimization of adding levels for the shaping property. Probiotic cell viability, rheology, texture, and digestion simulation were also compared. The results showed that the surviving amount of probiotics (107 CFU/g) in HPPS3:1-lpl@choc (add 15% HPPS3:1-lpl to the chocolate) was much higher than that in lpl@choc (102 CFU/g, without microencapsulation protection) after in-vitro digestion test, among which HPPS3:1-lpl@choc15 had the best shaping property. This research provides a new alternative strategy for utilizing porous starch carrier to encapsulate probiotics for melting 3D printing of chocolate (or other similar food materials) as personalized design.

Valorization of berry pomace for extraction of polyphenol compounds and its co-encapsulation with probiotic bacteria

Saskatoon berry pomace, an antioxidant rich byproduct, may be suitable for nutraceuticals and functional foods. The present study aimed to co-encapsulate polyphenol-rich berry extract with probiotics to utilize the polyphenolic compounds present in berry pomace. The major benefit of co-encapsulation is that polyphenolic compounds increase the survival characteristics of probiotic bacteria in gastrointestinal tract. To make the process cost-effective, a conventional solvent extraction method was used for extraction of polyphenolic compounds from berry pomace. Spray drying was used to co-encapsulate polyphenols and probiotics by using plant-based carrier materials (pea protein isolate with gum Arabic). Spray dried powder was evaluated for encapsulation efficiency, gastrointestinal stability, bio-accessibility index, along with functional, structural and thermal characteristics. Berry pomace was found to be a good source of TPC, DPPH and ABTS with 2.49 mg GAE/1 g, 4.48 mg QE/1 g and 2.96 mg QE/1 g, respectively. The encapsulation efficiency (retention of polyphenolics and bacteria in capsules) of polyphenolic compounds and probiotics was 72.6% and 94.4%, respectively. Probiotic cells encapsulated with polyphenolic compounds showed improved survival (9.08 log CFU/mL) during in vitro gastrointestinal digestion. The bio-accessibility of TPC was 63.6% after intestinal digestion. The spray dried powder was observed to possess good thermal stability but poor functional properties, thus limiting applications to products such as bakery goods, sports bars, cereals and other foods where dispersibility is not imperative. Therefore, co-encapsulation by spray drying method offers an efficient and cost-effective method for simultaneous delivery of bioactive compounds and probiotics to the gut, extending their benefits by this combination.

Survival of Probiotic Bacterial Cells in the Upper Gastrointestinal Tract and the Effect of the Surviving Population on the Colonic Microbial Community Activity and Composition.

Many health-promoting effects have been attributed to the intake of probiotic cells. However, it is important that probiotic cells arrive at the site of their activity in a viable state in order to exert their beneficial effects. Careful selection of the appropriate probiotic formulation is therefore required as mainly the type of probiotic species/strain and the administration strategy may affect survival of the probiotic cells during the upper gastrointestinal (GIT) passage. Therefore, the current study implemented Simulator of the Human Microbial Ecosystem (SHIME®) technology to investigate the efficacy of different commercially available probiotic formulations on the survival and culturability of probiotic bacteria during upper GIT passage. Moreover, Colon-on-a-Plate (CoaP™) technology was applied to assess the effect of the surviving probiotic bacteria on the gut microbial community (activity and composition) of three human donors. Significantly greater survival and culturability rates were reported for the delayed-release capsule formulation (>50%) as compared to the powder, liquid, and standard capsule formulations (<1%) (p < 0.05), indicating that the delayed-release capsule was most efficacious in delivering live bacteria cells. Indeed, administration of the delayed-release capsule probiotic digest resulted in enhanced production of SCFAs and shifted gut microbial community composition towards beneficial bacterial species. These results thus indicate that careful selection of the appropriate probiotic formulation and administration strategy is crucial to deliver probiotic cells in a viable state at the site of their activity (distal ileum and colon).

A potential therapeutic strategy of an innovative probiotic formulation toward topical treatment of diabetic ulcer: an in vivo study

BackgroundThe probiotic potential of Lacticacid bacteria has been studied in various medical complications, from gastrointestinal diseases to antibiotic resistance infections recently. Moreover, diabetic ulcer (DU) is known as one of the most significant global healthcare concerns, which comprehensively impacts the quality of life for these patients. Given that the conventional treatments of DUs have failed to prevent later complications completely, developing alternative therapies seems to be crucial.MethodsWe designed the stable oleogel-based formulation of viable probiotic cells, including Lactobacillus rhamnosus (L. rhamnosus), Lactobacillus casei (L. casei), Lactobacillus fermentum (L. fermentum), and Lactobacillus acidophilus (L. acidophilus) individually to investigate their effect on wound healing process as an in vivo study. The wound repair process was closely monitored regarding morphology, biochemical, and histopathological changes over two weeks and compared it with the effects of topical tetracycline as an antibiotic approach. Furthermore, the antibiofilm activity of probiotic bacteria was assessed against some common pathogens.ResultsThe findings indicated that all tested lactobacillus groups (excluded L. casei) included in the oleogel-based formulation revealed a high potential for repairing damaged skin due to the considerably more levels of hydroxyproline content of tissue samples along with the higher numerical density of mature fibroblasts cell and volume density of hair follicles, collagen fibrils, and neovascularization in comparison with antibiotic and control groups. L. acidophilus and L. rhamnosus showed the best potential of wound healing among all lactobacillus species, groups treated by tetracycline and control groups. Besides, L. rhamnosus showed a significant biofilm inhibition activity against tested pathogens.ConclusionsThis experiment demonstrated that the designed formulations containing probiotics, particularly L. acidophilus and L. rhamnosus, play a central role in manipulating diabetic wound healing. It could be suggested as an encouraging nominee for diabetic wound-healing alternative approaches, though further studies in detailed clinical trials are needed.

The Role of Proteomics in Identification of Key Proteins of Bacterial Cells with Focus on Probiotic Bacteria.

Probiotics can affect human health, keep the balance between beneficial and pathogenic bacteria, and their colonizing abilities enable the enhancement of the epithelial barrier, preventing the invasion of pathogens. Health benefits of probiotics were related to allergy, depression, eczema, cancer, obesity, inflammatory diseases, viral infections, and immune regulation. Probiotic bacterial cells contain various proteins that function as effector molecules, and explaining their roles in probiotic actions is a key to developing efficient and targeted treatments for various disorders. Systematic proteomic studies of probiotic proteins (probioproteomics) can provide information about the type of proteins involved, their expression levels, and the pathological changes. Advanced proteomic methods with mass spectrometry instrumentation and bioinformatics can point out potential candidates of next-generation probiotics that are regulated under pharmaceutical frameworks. In addition, the application of proteomics with other omics methods creates a powerful tool that can expand our understanding about diverse probiotic functionality. In this review, proteomic strategies for identification/quantitation of the proteins in probiotic bacteria were overviewed. The types of probiotic proteins investigated by proteomics were described, such as intracellular proteins, surface proteins, secreted proteins, and the proteins of extracellular vesicles. Examples of pathological conditions in which probiotic bacteria played crucial roles were discussed.

The Impact of Annealing Methods on the Encapsulating Structure and Storage-Stability of Freeze-Dried Pellets of Probiotic Bacteria

ObjectiveThis paper investigates the critical role of material thickness in freeze-dried pellets for enhancing the storage stability of encapsulated bacteria. Freeze dried material of varying thicknesses obtained from different annealing durations is quantified using Scanning Electron Microscopy (SEM) and X-ray microtomography (μCT), the material thickness is then correlated to the storage stability of the encapsulated cells.MethodsA formulation comprising of sucrose, maltodextrin, and probiotic cells is quenched in liquid nitrogen to form pellets. The pellets undergo different durations of annealing before undergoing freeze-drying. The material thickness is quantified using SEM and μCT. Storage stability in both oxygen-rich and oxygen-poor environments is evaluated by measuring CFU counts and correlated with the pellet structure.ResultsThe varying annealing protocols produce a range of material thicknesses, with more extensive annealing resulting in thicker materials. Storage stability exhibits a positive correlation with material thickness, indicating improved stability with thicker materials. Non-annealed pellets exhibit structural irregularities and inconsistent storage stability, highlighting the impracticality of avoiding annealing in the freeze-drying process.ConclusionsExtensive annealing not only enhances the storage stability of probiotic products but also provides greater control over the freeze-drying process, ensuring homogeneous and reproducible products. This study underscores the importance of material thickness in freeze-dried pellets for optimizing storage stability for probiotic formulations, and emphasize the necessity of annealing as a critical step in freeze-drying quenched pellets to achieve desired structural and stability outcomes.

Anticandidal Activity of Various Probiotic Lactobacillus Strains and Their Efficacy Enhanced by Prebiotic Supplementation.

Probiotics and prebiotics have been considered as alternative approaches for promoting health. This study aimed to investigate the anticandidal potential of various probiotic Lactobacillus strains and their cell-free supernatants (CFSs). The study assessed the impact of inulin and some fruitsas prebiotics on the growth of selected probiotic strains in relation to their anticandidal activity, production of short-chain fatty acids,total phenolic content, and antioxidant activity. Results revealed variations in anticandidal activity based on the specific strains and forms of probiotics used. Non-adjusted CFSs were the most effective against Candida strains, followed by probiotic cells and adjusted CFSs (pH 7). Lacticaseibacillus rhamnosus SD4, L. rhamnosus SD11 and L. rhamnosus GG displayed the strongest anticandidal activity. Non-adjusted CFSs from L. rhamnosus SD11, L. rhamnosus SD4 and L. paracasei SD1 exhibited notable anticandidal effects. The adjusted CFSs of L. rhamnosus SD11 showed the highest anticandidal activity against all non-albicans Candida (NAC) strains, whereas the others were ineffective. Supplementation of L. rhamnosus SD11 with prebiotics, particularly 2% (w/v) mangosteen, exhibited positive results in promoting probiotic growth, short-chain fatty acids production, total phenolic contents, and antioxidant activity, and the subsequent enhancing anticandidal activity against both C. albicans and NAC strains compared to conditions without prebiotics. In conclusion, both live cells and CFSs of tested strains, particularly L. rhamnosus SD11, exhibited the best anticandidal activity. Prebiotics supplementation, especially mangosteen, enhanced probiotic growth and beneficial metabolites against Candida growth. These finding suggested that probiotics and prebiotic supplementation may be an effective alternative treatment for Candida infections.

Exploring prebiotic properties and its probiotic potential of new formulations of soy milk-derived beverages.

The food and beverage industry has shown a growing interest in plant-based beverages as alternatives to traditional milk consumption. Soy milk is derived from soy beans and contains proteins, isoflavones, soy bean oligosaccharides, and saponins, among other ingredients. Because of its high nutritive value and versatility, soy milk has gained a lot of attention as a functional food. The present work aims to explore the prebiotic properties and gastrointestinal tolerance potential of new formulations of soy milk-derived drinks to be fermented with riboflavin-producing probiotic Lactiplantibacillus plantarum MTCC (Microbial Type Culture Collection and Gene Bank) 25432, Lactiplantibacillus plantarum MTCC 25433, and Lactobacillus acidophilus NCIM (National Collection of Industrial Microorganisms) 2902 strains. The soy milk co-fermented beverage showed highest PAS (1.24 ± 0.02) followed by soy milk beverages fermented with L. plantarum MTCC 25433 (0.753 ± 0.0) when compared to the commercial prebiotic raffinose (1.29 ± 0.01). The findings of this study suggested that the soy milk beverages exhibited potent prebiotic activity, having the ability to support the growth of probiotics, and the potential to raise the content of several bioactive substances. The higher prebiotics activity score showed that the higher the growth rate of probiotics microorganism, the lower the growth of pathogen. For acidic tolerance, all fermented soy milk managed to meet the minimal requirement of 106 viable probiotic cells per milliliter at pH 2 (8.13, 8.26, 8.30, and 8.45 logs CFU/mL, respectively) and pH 3.5 (8.11, 8.07, 8.39, and 9.01 log CFU/mL, respectively). The survival rate of soy milk LAB isolates on bile for 3 h ranged from 84.64 to 89.60%. The study concluded that lactobacilli could thrive in gastrointestinal tract. The sensory evaluation scores for body and texture, color, flavor, and overall acceptability showed a significant difference (p < 0.05) between the fermented probiotic soy milk and control samples. Soy milk fermented with a combination of L. plantarum MTCC 25432 & MTCC 25433 demonstrated the highest acceptability with the least amount of beany flavor. The findings of the study suggest soy milk's potential in plant-based beverage market.

Bifidobacterium animalis subsp. lactis HN019 live probiotics and postbiotics: production strategies and bioactivity evaluation for potential therapeutic properties.

Introduction: B. animalis subsp. lactis HN019 is a commercially available well-characterized probiotic with documented effects on human health, such as the ability to enhance the immune function and to balance the intestinal microbiome. Therefore, optimizing the manufacturing process to improve sustainability, increasing biomass yields and viability, and avoiding animal -derived nutrients in the medium to meet vegan consumer's needs, is currently of interest. Besides the established use of live probiotic cells, alternative supplements indicated as postbiotics, like non-viable cells and/or probiotics derived bioactive molecules might be considered as potential next generation biotherapeutics. In fact, advantages of postbiotics include fewer technological limitations, such as easier production processes and scale-up, and even higher specificity. Methods: In this work, medium design together with different fermentation strategies such as batch, fed-batch and in situ product removal on lab-scale bioreactors were combined. Medium pretreatment by ultrafiltration and protease digestion was performed to reduce polysaccharidic contaminants and facilitate the purification of secreted exopolysaccharides (EPS). The latter were isolated from the fermentation broth and characterized through NMR, GC-MS and SEC-TDA analyses. The expression of TLR-4, NF-kb and IL-6 in LPS challenged differentiated CaCo-2 cells treated with EPS, live and heat-killed B. lactis cells/broth, was evaluated in vitro by western blotting and ELISA. Zonulin was also assessed by immunofluorescence assays. Results and Discussion: The titer of viable B. lactis HN019 was increased up to 2.9 ± 0.1 x 1010 on an animal-free semidefined medium by applying an ISPR fermentation strategy. Medium pre-treatment and a simple downstream procedure enriched the representativity of the EPS recovered (87%), the composition of which revealed the presence of mannuronic acid among other sugars typically present in polysaccharides produced by bifidobacteria. The isolated EPS, live cells and whole heat inactivated broth were compared for the first up to date for their immunomodulatory and anti-inflammatory properties and for their ability to promote intestinal barrier integrity. Interestingly, EPS and live cells samples demonstrated immune-stimulating properties by downregulating the expression of TLR-4 and NF-kb, and the ability to promote restoring the integrity of the intestinal barrier by up-regulating the expression of zonulin, one of the tight junctions forming proteins. Postbiotics in the form of heat killed broth only reduced NF-kb expression, whereas they did not seem effective in the other tested conditions.

Label-Free Monitoring of Coculture System Dynamics: Probing Probiotic and Cancer Cell Interactions via Infrared Spectroscopic Imaging.

Evaluating the dynamic interaction of microorganisms and mammalian cells is challenging due to the lack of suitable platforms for examining interspecies interactions in biologically relevant coculture conditions. In this work, we demonstrate the interaction between probiotic bacteria (Lactococcus lactis and Escherichia coli) and A498 human cancer cells in vitro, utilizing a hydrogel-based platform in a label-free manner by infrared spectroscopy. The L. lactis strain recapitulated in the compartment system secretes polypeptide molecules such as nisin, which has been reported to trigger cell apoptosis. We propose a mid-infrared (IR) spectroscopic imaging approach to monitor the variation of biological components utilizing kidney cells (A498) as a model system cocultured with bacteria. We characterized the biochemical composition (i.e., nucleic acids, protein secondary structures, and lipid conformations) label-free using an unbiased measurement. Several IR spectral features, including unsaturated fatty acids, β-turns in protein, and nucleic acids, were utilized to predict cellular response. These features were then applied to establish a quantitative relationship through a multivariate regression model to predict cellular dynamics in the coculture system to assess the effect of nisin on A498 kidney cancer cells cocultured with bacteria. Overall, our study sheds light on the potential of using IR spectroscopic imaging as a label-free tool to monitor complex microbe-host cell interactions in biological systems. This integration will enable mechanistic studies of interspecies interactions with insights into their underlying physiological processes.

Delivery of Probiotic-Loaded Microcapsules in the Gastrointestinal Tract: A Review.

Probiotics are live microorganisms that inhabit the gastrointestinal tract and confer health benefits to consumers. However, a sufficient number of viable probiotic cells must be delivered to the specific site of interest in the gastrointestinal tract to exert these benefits. Enhanced viability and tolerance to sublethal gastrointestinal stress can be achieved using appropriate coating materials and food matrices for orally consumed probiotics. The release mechanism and interaction of probiotic microcapsules with the gastrointestinal tract have been minimally explored in the literature to date. To the authors' knowledge, no review has been published to discuss the nature of release and the challenges in the targeted delivery of probiotics. This review addresses gastrointestinal-related complications in the formulation of targeted delivery and controlled release of probiotic strains. It investigates the impacts of environmental stresses during the transition stage and delivery to the target region in the gastrointestinal tract. The influence of factors such as pH levels, enzymatic degradation, and redox conditions on the release mechanisms of probiotics is presented. Finally, the available methods to evaluate the efficiency of a probiotic delivery system, including in vitro and in vivo, are reviewed and assessed. The paper concludes with a discussion highlighting the emerging technologies in the field and emphasising key areas in need of future study.

Bioconversion of mangosteen pericarp extract juice with bacterial cellulose production and probiotic integration: A promising approach for functional food supplement development

This study optimized bioconversion process of mangosteen pericarp extract juice (MPEj) with a single-culture of Komagataeibacter xylinus TISTR 1061, a bacterial cellulose-producing strain and a co-culture with the probiotic Lactiplantibacillus plantarum. K. xylinus TISTR 1061 was identified as the optimal bacterial cellulose-producing strain for fermenting MPEj. Optimal conditions were determined to be 20% (v/v) MPEj supplemented with 7% (w/v) sucrose and 0.2% (w/v) ammonium dihydrogen phosphate as carbon and nitrogen sources, respectively. The growth profile of single K. xylinus was not significantly different from those of the co-culture. They showed good adaptability and cohabited during fermentation without antagonistic effects. Dynamic changes in total titratable acidity (TTA), total soluble solids (TSS), and pH during fermentation were relatively similar for both single- and co-cultures. Post-fermented MPEj with co-culture exhibited significantly higher bioactive activities, particularly antioxidant properties. Significant variations in metabolite concentrations, including 2-aminobutyrate, valine, xylitol, propionate, isoleucine, glucose, and glycerol, were observed. These compounds exhibited significantly higher average concentrations after fermentation, highlighting their potential as biomarkers distinguishing pre- and post-fermentation MPEj. Bacterial cellulose (BC) from single culture exhibited greater thickness and weight, with a more complex and branched structure, while that from co-culture was infused with probiotic cells, showing less complexity allowing the BC from co-culture demonstrated low hardness and chewiness values. The properties of both bioconverted MPEj and BC obtained as observed offer potential for innovative functional drinks and/or dietary supplements with enhanced health benefits.