Protection Of Probiotics Research Articles

Co-encapsulation of probiotics with acylglycerols in gelatin-gum arabic complex coacervates: Stability evaluation under adverse conditions

Co-encapsulation of acylglycerols and probiotics may improve the resistance of probiotics to adverse conditions. In this study, three probiotic microcapsule models were constructed using gelatin (GE)-gum arabic (GA) complex coacervate as wall material: microcapsules containing only probiotics (GE-GA), microcapsules containing triacylglycerol (TAG) oil and probiotics (GE-T-GA) and microcapsules containing diacylglycerol (DAG) oil and probiotics (GE-D-GA). The protective effects of three microcapsules on probiotic cells under environmental stresses (freeze-drying, heat treatment, simulated digestive fluid and storage) were evaluated. The results of cell membrane fatty acid composition and Fourier transform infrared (FTIR) spectroscopy revealed that GE-D-GA could improve the fluidity of cell membrane, maintain the stability of protein and nucleic acid structure, and decrease the damage of cell membrane. These characteristics supported the high freeze-dried survival rate (96.24 %) of GE-D-GA. Furthermore, regardless of thermotolerance or storage, GE-D-GA showed the best cell viability retention. More importantly, GE-D-GA provided the best protection for probiotics under simulated gastrointestinal conditions, as the presence of DAG reduced cell damage during freeze-drying and the degree of contact between probiotics and digestive fluids. Therefore, co-microencapsulation of DAG oil and probiotics is a promising strategy to resist adverse conditions.

Lactobacillus rhamnosus GG alleviates colitis caused by chemotherapy via biofilm formation.

Severe colitis is a common side effect of chemotherapy in cancer patients. In this study, we attempted to enhance the viability of probiotics in a gastric acid environment and improve the colitis induced by dextran sulfate sodium (DSS) and docetaxel. We purified Lactobacillus from yogurt and estimated their growth at pH 6.8 and pH 2.0. In the further investigation, the bacterial biofilm formation was used to define the mechanism by which administration of Lactobacillus rhamnosus (LGG) via oral gavage alleviates the colitis and intestine permeability of the mice induced by DSS and docetaxel. The potential benefit of probiotics on the treatment of breast cancer metastasis has been assessed as well. Lactobacillus from yogurt growth was unexpectedly faster in the pH 2.0 than in the neutral pH medium during the first hour. LGG administered in the fasting state via oral gavage significantly improved the preventive effect in the colitis caused by DSS and docetaxel. LGG reduced the permeability of the intestine and decreased the expression of proinflammatory cytokines, TNF-α, IL-1β, and IL-6, in colitis by biofilm formation. Increasing the docetaxel dose may reduce breast tumor growth and metastasis in the lung but did not benefit survival due to severe colitis. However, the LGG supplement significantly improved the survival of tumor-bearing mice following a high dose of docetaxel treatment. Our findings provide new insights into the potential mechanism of probiotic protection of the intestine and provide a novel therapeutic strategy to augment the chemotherapeutic treatment of tumors.

Green tea polysaccharide conjugates and gelatin enhanced viability of L. acidophilus by layer-by-layer encapsulation

Adverse environments such as the gastrointestinal tract can affect the efficacy of probiotics. Therefore, the adequate protection of probiotics and their total effect exertion are probiotics application research hotspots. Green tea polysaccharide conjugate (gTPC), an anionic polymer, and gelatin as the encapsulation materials, and layer-by-layer self-assembly technology as the method, were applied to encapsulate Lactobacillus acidophilus. Microscopic results showed that with the increase in the number of deposited layers, the encapsulation was more complete. After continuous exposure to simulated gastrointestinal fluid for 120 min, the survival rate of unencapsulated cells was 53.26%, and encapsulated cells with different deposited layers were 66.00%, 71.99%, 76.94%, and 79.89%, respectively. In addition, Gel/gTPC bilayers can efficiently improve cell thermotolerance and freezing resistance. Results indicated that the more the number of Gel/gTPC bilayers, the more significant the protection is. Therefore, gTPC and Gel are viable materials to protect probiotics through microencapsulation by Layer-by-layer self-assembly.

Cruciferin improves stress resistance and simulated gastrointestinal survival of probiotic Limosilactobacillus reuteri in the model encapsulation system

Encapsulation is a viable strategy to improve the stability and survival of probiotics during processing, storage, and consumption. Cruciferin, a major canola protein with high denaturation temperature and resistance to gastric degradation, has potential for encapsulation and protection of probiotics against harsh conditions in processing and gastrointestinal tract. Cruciferin/alginate capsules were fabricated to encapsulate probiotics, and were characterized using confocal and scanning electron microscopy (SEM). The bacterial viability was studied during storage, processing, and gastro-intestinal transit. Limosilactobacillus reuteri TMW 1.656 was encapsulated in spherical cruciferin/alginate capsules (2.2 ± 0.1 mm) prepared using an extrusion method. SEM images of the capsules showed that the bacteria were entrapped within the porous structure which was formed by the complexation of cruciferin and alginate. The confocal microscopy images confirmed that cruciferin and alginate were homogeneously distributed throughout the capsules. The shelf life of the bacteria in the presence of cruciferin and alginate increased up to 8 weeks at 4 °C, while unencapsulated (free) bacteria lost their viability after 2 weeks storage. The heat resistance of encapsulated bacteria exposed to 65 °C and 70 °C was improved by up to ∼ 4 and 2 log cycles, respectively, compared to unencapsulated bacteria. Encapsulation also protected L. reuteri against gastric low pH and enzymes; the viability was 3 logs higher when compared to unencapsulated bacteria. The capsules were degraded in simulated intestinal fluid, leading to the release of the encapsulated bacteria, whereas the wall materials increased the resistance of released bacteria to bile salts. Comparison between the viability of unencapsulated bacteria in presence of cruciferin/alginate mixtures and bacteria encapsulated in the capsules revealed that capsule formation provided physical barriers to the harsh conditions and played a key role in the protection of bacteria. This study showed that cruciferin/alginate capsules are capable to improve stability and shelf life of Limosilactobacillus reuteri.

Microencapsulation by a Spray Drying Approach to Produce Innovative Probiotics-Based Products Extending the Shelf-Life in Non-Refrigerated Conditions

Recently, there has been a growing interest in producing functional foods containing encapsulated probiotic bacteria due to their positive effects on human health. According to their perceived health benefits, probiotics have been incorporated into a range of dairy products, but the current major challenge is to market new, multicomponent probiotic foods and supplements. Nevertheless, only a few products containing encapsulated probiotic cells can be found as non-refrigerated products. In this work, spray drying technology was investigated in order to produce an innovative nutraceutical formulation based on lactic acid bacteria (LAB), and was able to ensure a good storage stability of probiotics (no less than 109 CFU/cps) in non-refrigerated conditions. Probiotic-loaded microparticles from spray drying experiments were produced under different conditions and compared by thermogravimetric analysis (TGA), scanning electron microscopy (SEM), and the enumeration of the number of viable cells in order to identify the formulation exhibiting the most promising characteristics. Results from the dissolution test revealed that the optimized formulation provides a suitable amount of living cells after digestion of microparticles stored for 12 months at room temperature and confirmed that the microencapsulation process by spray drying ensures a good protection of probiotics for nutraceutical purposes.

Microencapsulation of Probiotics by Oil-in-Water Emulsification Technique Improves Cell Viability under Different Storage Conditions.

Probiotics are associated with health benefits to the host. However, their application can be limited due to a decrease in cell viability during processing, storage, and passage through the gastrointestinal tract. Microencapsulation is a simple and efficient alternative to improve the physical protection and stability of probiotics. The present study aimed to produce and characterize alginate or gelatin-based microparticles containing Lactobacillus acidophilus NRRL B-4495 or Lactiplantibacillus plantarum NRRL B-4496 by oil-in-water (O/W) emulsification and to evaluate the stability under storage conditions. The results showed that L. acidophilus and L. plantarum encapsulated in gelatin (LAEG and LPEG) presented diameters of 26.08 ± 1.74 μm and 21.56 ± 4.17 μm and encapsulation efficiencies of 89.6 ± 4.2% and 81.1 ± 9.7%, respectively. However, those encapsulated in alginate (LAEA and LPEA) showed an encapsulation efficiency of <1.0%. Furthermore, LAEG was stable for 120 days of storage at 5 °C and 25 °C. Therefore, encapsulation in gelatin by O/W emulsification is a promising strategy for protecting and stabilizing probiotic bacteria, enabling future application in foods.

Improving the viability of Lactobacillus plantarum LP90 by carboxymethylated dextran-whey protein conjugates: The relationship with glass transition temperature

Microcapsules can provide better protection for probiotics during freeze-drying and gastrointestinal digestion. This study aims to investigate the effect of microencapsulation using the product of the Maillard reaction of whey protein and carboxymethylated dextran (CM. dex) on the viability of Lactobacillus plantarum LP90 ( L p ) as related to freeze-drying, gastrointestinal tolerability and storage stability. The results demonstrate that the glycosylation product was successfully obtained by the dry heating method, and the product was verified by SDS–PAGE, infrared spectroscopy (FTIR), and intrinsic fluorescence spectroscopy. The grafting degree of whey protein and CM. dex, as measured by the OPA method, was 53.87%, 57.60%, and 61.21% at 1, 3, and 5 days of the Maillard reaction, respectively. With increasing Maillard reaction time, the glass transition temperature ( T g ) of conjugates increased, and WP–CD 5d had the highest T g , which was 156.31 °C. At the same time, WP–CD 5d had the best protection effect under freeze-drying, and the survival rate was 87.37%. Microencapsulated L p had better tolerance over 5 h of simulated gastrointestinal juice treatment fluid. L p had the best storage stability protected by WP-CD 5d , for which k was 0.102. • Carboxymethylated dextran was applied on microencapsulation of probiotics. • WP-CD 5d had the highest glass transition temperature ( T g ), which was 156.31 °C. • Microencapsulated LP90 decreased slightly in gastrointestinal simulant. • L p had the best storage stability protected by WP-CD 5d , which k was 0.102.

Effect of pectic oligosaccharide on probiotic survival and physicochemical properties of hydrogel beads for synbiotic encapsulation of Lactobacillus bulgaricus

The present study was carried out to evaluate the effects of different concentrations of pectic oligosaccharide (POS) on the physicochemical properties of Lactobacillus bulgaricus (L. bulgaricus) loaded-synbiotic hydrogel beads and the survival of L. bulgaricus. Low methoxyl (LM) pectin-resistant starch (RS) hydrogel beads, named RS beads, were coated/filled with different concentrations of POS: 0.8% (RS-POS0.8 beads), 1.0% (RS-POS1.0 beads), and 1.2% (RS-POS1.2 beads). POS treatment resulted in smooth beads with less surface dent by hydrogen bonding between RS beads and POS, which was confirmed by SEM and XRD analyses. The encapsulation efficiency of RS beads and RS-POS beads ranged from 90.40% to 92.03%. In simulated gastric fluid (SGF) and bile salt solution, free L. bulgaricus completely died but L. bulgaricus encapsulated in RS-POS1.2 beads only slightly decreased. This indicates that RS-POS1.2 beads exhibited higher protective effects on L. bulgaricus under SGF condition and bile salts. Furthermore, considering the relatively higher storage stability of L. bulgaricus loaded in RS-POS1.2 beads, the present results suggest that RS beads coated/filled with POS could be used for synbiotic encapsulation, protection, and delivery of probiotics.

Measuring of the heifers ovaries and probiotic defence of the mucous membranes

The results of the influence of complex probiotic defence of the mucous membranes of the reproductive system and intestines on the development of the corpus luteum of the cycle in heifers have been presented. By the method of differential palpation diagnostics in vivo, it was reliably examined that in the experimental group after carrying out procedures for the normalization of mucous membranes with the multicomponent probiotic drug “Multibacterin Veterinary Ba + La”, the number of heifers with high-quality of corpus luteum the ovaries was 48.21% more, and the number of cases of premature luteolysis was 12.50 % less (P &lt; 0.01). The level of pregnancy in heifers in the experiment was on average 60.38 %, in the control – 46.94 %, which confirmed the positive effect of the probiotic protection of the mucous membranes on increasing the embryo survival by optimizing the conditions for embryonic nutrition on the surface of the endometrium.

Probiotics protect against hepatic steatosis in tris (2-chloroethyl) phosphate-induced metabolic disorder of mice via FXR signaling.

Tris (2-chloroethyl) phosphate (TCEP), the most widely useful and most frequently detective organophosphate flame retardants in environment, has been shown potential relationship with adolescent weight. Probiotics is an effective therapy for metabolic diseases such as obesity and NAFLD with gut microbiota dysregulation. This study aims to explore the protective effects of probiotics against lipid metabolic disorder induced by chronic TCEP exposure and demonstrate the mechanism of this event. The data showed that dietary complex probiotics supplement attenuated TCEP-induced obesity, hyperlipidemia, liver dysfunction, and hepatic steatosis. In addition, dietary complex probiotics suppressed TCEP-promoted ileal FXR signaling, and upregulated hepatic FXR/SHP pathway inhibited by TCEP. Moreover, dietary complex probiotics stimulated PPARα-mediated lipid oxidation and suppressed SREBP1c/PPARγ-mediated lipid synthesis via regulation of FXR signaling. Therefore, this study indicates that dietary complex probiotics could protect against hepatic steatosis via FXR-mediated signaling pathway in TCEP-induced metabolism disorder in mice, resulting in attenuation of systemic lipid accumulation.

Enhancing viability of Lactobacillus plantarum encapsulated by alginate-gelatin hydrogel beads during gastrointestinal digestion, storage and in the mimic beverage systems

To improve the viability of Lactobacillus plantarum (P) during digestion and storage, the probiotics were encapsulated by alginate (ALG) and alginate-gelatin (ALG-GE) hydrogels beads. ALG-P-GE showed much better physicochemical properties than ALG-P. The scanning electron microscopy (SEM) results validated the incorporation of bacterial cells into the beads. ALG-P-GE exhibited good encapsulation efficiency of 97.7 %, and the storage and thermal stability of probiotic were increased by 15 % and 8 %, respectively, when comparing with ALG-P. ALG-P-GE beads could protect the probiotics from inactivation in simulated gastric fluid and then release it in simulated intestinal fluid. The protective mechanism of ALG-GE for probiotics was further studied by fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD) and found that ALG and GE can form gel network through hydrogen bonding and electrostatic interactions. In the mimic beverage systems, ALG-P-GE beads could protect the encapsulated probiotics and increase its viability. The storage, thermal, and digestion stability of encapsulated probiotic were significantly increased and showed high viability in the mimic beverage systems. ALG-P-GE beads have great potential for the protection and delivery of probiotics in food systems.

Probiotics viability in frozen food products

Improving gastrointestinal flora with probiotics is a promising method of treating and preventing many diseases. However, in food industry, introduction of probiotics is limited by the shelf life of products, as well as by the reduction of bacterial viability during storage. Thus, introduction of probiotics into foods to be frozen is a promising method to extend the shelf life of probiotic products. However, the storage of living cells in frozen products involves their damage during freezing and subsequent storage at temperatures below 0 °C. Therefore, some technological solutions are required to protect probiotics during storage at sub-zero. Today, encapsulation, addition of cryoprotectants and prebiotics, as well as adaptation of microorganisms are widely used techniques of protecting probiotics. The combination of various protection methods is a promising way to improve the viability of probiotics during storage. Studies aimed at investigating various combinations of hurdle methods and their effect on bacterial survival are of great interest.This review compiles the most common methods of preserving probiotics in frozen foods, describing their effects, variety of approaches, and effectiveness. The main mechanisms of bacterial cell damage by freezing are described. Further trends in probiotic protection in frozen foods are also discussed.

Lactobacillus plantarum 21805 encapsulated by whey protein isolate and dextran conjugate for enhanced viability

Probiotics incorporated into food products are often insufficiently potent because of unfavorable interference during processing. This study addressed these issues by encapsulating Lactobacillus plantarum into whey protein isolate (WPI) and dextran (DX) conjugates using an encapsulator. The objective of this work is to investigate the effect of encapsulation using WPI and DX conjugates on the viability of Lactobacillus plantarum. The formation of the conjugates was confirmed with absorbance of the intermediate- and late-stage products, degree of grafting, browning index, and gel electrophoresis. Extending the heating time from 1 to 5 h increased the content of high molecular weight conjugates. These conjugates provided better protection of probiotics under processing stress conditions, gastrointestinal conditions, and storage. Furthermore, encapsulated L. plantarum exhibited a reduction of only 0.33 log CFU/mL after 90 d of storage at 4 °C. Thus, microencapsulation of probiotics can enhance the viability of functional food products and facilitate corresponding research.

Functional fermented meat products with probiotics—A review

Fermentation has been an important strategy in the preservation of foods. The use of starter cultures with probiotic activity has gained the attention of researchers to produce functional fermented meat products. This review aims to overview the main strengths, weakness, opportunities and threats of fermented meat products with probiotics. Fermented meat products can be considered as a relevant matrix for the delivery of probiotics with potential health benefits. Moreover, fermented meat products produced by traditional methods are sources of probiotics that can be explored in the production of functional meat products. However, some barriers are limit the progression with these products: the complex selection process to obtain new and tailored probiotic strains, the current perception of healthiness associated with meat and meat products, and the limited application of probiotic to fermented sausages. Promising opportunities to improve the value of functional fermented meat products have been developed by exploring new meat products as functional fermented foods, improving the protection of probiotics with microencapsulation and improving the quality of meat product (reducing nitrate and nitrate salts, adding dietary fibre, and exploring the inherent antioxidant and cardioprotective activity of meat products). Attention to potential threats is also indicated such as the unclear future changes in meat and meat products consumption due to changes in consumer preferences and the presence of competitors (dairy, fruit and vegetable-based products, for instance) in more advanced stages of development and commercialization. SIGNIFICANCE AND IMPACT OF STUDY: This review provides an overview of the Strengths, Weakness, Opportunities and Threats related to the development of functional fermented meat products with probiotics. Internal and external factors that explain the current scenario and strategies to advance the production are highlighted.

Activation of a passive, mesoporous silica nanoparticle layer through attachment of bacterially-derived carbon-quantum-dots for protection and functional enhancement of probiotics.

Probiotic bacteria employed for food supplementation or probiotic-assisted antibiotic treatment suffer from passage through the acidic gastro-intestinal tract and unintended killing by antibiotics. Carbon-quantum-dots (CQDs) derived from bacteria can inherit different chemical groups and associated functionalities from their source bacteria. In order to yield simultaneous, passive protection and enhanced, active functionality, we attached CQDs pyrolytically carbonized at 220 ​°C from Lactobacillus acidophilus or Escherichia coli to a probiotic strain (Bifidobacterium infantis) using boron hydroxyl-modified, mesoporous silica nanoparticles as an intermediate encapsulating layer. Fourier-transform-infrared-spectroscopy, X-ray-photoelectron-spectroscopy and scanning-electron-microscopy were employed to demonstrate successful encapsulation of B. infantis by silica nanoparticles and subsequent attachment of bacterially-derived CQDs. Thus encapsulated B. infantis possessed a negative surface charge and survived exposure to simulated gastric fluid and antibiotics better than unencapsulated B. infantis. During B. infantis assisted antibiotic treatment of intestinal epithelial layers colonized by E. coli, encapsulated B. infantis adhered and survived in higher numbers on epithelial layers than B. infantis without encapsulation or encapsulated with only silica nanoparticles. Moreover, higher E. coli killing due to increased reactive-oxygen-species generation was observed. In conclusion, the active, protective encapsulation described enhanced the probiotic functionality of B. infantis, which might be considered as a first step towards a fully engineered, probiotic nanoparticle.

Probiotic encapsulation in water-in-oil high internal phase emulsions: Enhancement of viability under food and gastrointestinal conditions

Polyglycerol polyricinoleate (PGPR) was used as the oil-soluble surfactant and beeswax was used as the oil phase to formulate a water-in-oil (W/O) high internal phase emulsions (HIPEs) for the encapsulation and protection of probiotics. The physicochemical properties of the W/O HIPEs and the survival of the encapsulated probiotics when exposed to acidic conditions and in vitro digestion were investigated. The viability of the probiotics decreased slightly when exposed to high-speed shearing. The rheological analysis, microstructural images, physicochemical stability showed that the W/O HIPEs remained relatively stable. The survival of the probiotics loaded in the SK-HIPEs (prepared with skim milk) was much higher than in the NS-HIPEs (prepared with normal saline) during storage at 4 °C. An in vitro gastrointestinal model showed that encapsulation of the probiotics enhanced their survival. This study provides useful insights into the utilization of W/O HIPEs to improve the efficacy of probiotics in the food industry.

Effects of protein fibrillation and antioxidants on probiotic survival during ambient storage

Drying of probiotic cultures is the main form of stabilization but dry probiotics face oxidative stress. The addition of antioxidants has been employed for protection of probiotics against oxidative stress, but results on the addition of antioxidants remain inconclusive. In this work, matrices of whey protein isolate fibrils (WPIF) with epigallocatechin gallate (EGCG) or glutathione (GSH) were used. Probiotic viability during ambient storage decreased in the order of WPIF > WPIF + EGCG > WPIF + GSH > WPI > WPI + EGCG > WPI + GSH. The improved protection of WPIF might be explained by its better cell encapsulation and the high antioxidant activity of WPIF. Both antioxidants accelerated the death of probiotic, which might be related with the antimicrobial activity or the cytotoxicity of the reaction products. This study proposed an excellent wall material of amyloid fibrils for probiotic protection during ambient storage, and questioned the “common sense” that antioxidants protect probiotics from oxidative stress.

Carboxymethyl konjac glucomannan-chitosan complex nanogels stabilized double emulsions incorporated into alginate hydrogel beads for the encapsulation, protection and delivery of probiotics

In this study, we developed (W1/O/W2) double emulsions encapsulated by calcium-alginate hydrogel beads system (ACGs) for intestinal-targeted delivery of probiotics (Lactobacillus reuteri). Firstly, the carboxymethyl konjac glucomannan-chitosan (CMKGM-CS) nanogels were successfully fabricated by EDC/NHS initiated crosslinking, as concluded from the TEM images, FTIR spectra, XRD, etc. Then, double emulsions were prepared and encapsulated with various concentrations of alginate to form ACGs hydrogel beads. In vitro probiotic release experiments showed the lyophilized ACG-2 and ACG-3 hydrogel beads had a sustained release in simulated intestinal fluid (SIF), the viability of cells exceeded 107 CFU/mL at 6 h. The lyophilized ACG-3 hydrogel beads exhibited the viable release amount of 8.4 × 107 CFU/mL after storage for 90 d at 4 °C. Besides, the alginate concentration in the ACGs hydrogel beads influences the swelling behavior and structure of hydrogel beads by affecting the hydrogen bonds between alginate and CMKGM-CS, thereby mediating the release of probiotics.

Research Progress in the Application of Electrospinning Technology in the Protection of Active Ingredients and Probiotics

Research Progress in the Application of Electrospinning Technology in the Protection of Active Ingredients and Probiotics

In vitro digestion of sodium alginate/pectin co-encapsulated Lactobacillus bulgaricus and its application in yogurt bilayer beads

The purpose of this study was to prepare sodium alginate (SA)/pectin (PE) hydrogel microspheres using the extrusion method to encapsulate Lactobacillus bulgaricus. Microscopic observation showed that the beads were spherical with a smooth and uniform surface. For microspheres with a diameter range of 140–156 μm, the encapsulation efficiency reached 85.67%. After simulating saliva, gastric juice, and intestinal juice, the activity of the microcapsules was estimated to be 5.78 × 104 log colony forming unit (CFU)/mL. These data show that the use of SA and PE encapsulated probiotics exhibit enhanced viability. In addition, double-layer beads containing probiotic microspheres and yogurt were prepared, and physical and chemical analysis was performed using scanning electron microscopy, Fourier-transform infrared spectroscopy, and differential scanning calorimetry. Texture and sensory property analysis revealed that the beads had good elasticity, chewiness, and high commercial value. Collectively, these findings indicate that SA and PE can be used for the encapsulation, protection, and gastrointestinal delivery of probiotics. Moreover, these microcapsules exhibit good stability in vitro and improve yogurt characteristics by increasing the survival rate of encapsulated probiotics, thus demonstrating their commercial application potential.