Lactiplantibacillus plantarum MIUG BL21 paraprobiotics: Evidences on inactivation kinetics and their potential as cytocompatible and antitumor alternatives.

Two new -biotics concepts, such as paraprobiotics and postbiotics were introduced, with beneficial effects beyond the viability of probiotic. In this study, the effect of individual (thermal, ohmic heating, high pressure, and ultrasound) and combined (ohmic, high pressure and ultrasound in combination with heating) treatments on the inactivation kinetics of Lactiplantibacillus plantarum was investigated. Different inactivation rates were obtained, up to 8.18 after 10 min at 90 °C, 2.07 after 15 min at a voltage gradient of 20 V/cm, 6.62 after 10 min at 600 MPa and 3.6 after ultrasound treatment for 10 min at 100 % amplitude. The experimental data were fitted to Weibullian model proposed by Peleg, allowing to estimate the inactivation rate coefficient (b) and the shape of the inactivation curves (n). At lower concentration, the samples showed both cytocompatibility and antiproliferative effect, stimulating the cell proliferation on both murine fibroblast and human colorectal adenocarcinoma cell lines.

Upgrading the Functional Potential of Apple Pomace in Value-Added Ingredients with Probiotics.

Emerging customized designs to upgrade the functional potential of freeze-dried apple pomace was used in this study, in order to transform the industrial by-products into ingredients containing probiotics, for a better and healthier food composition. The freeze-dried apple pomace was analyzed for free and bounded phenolic contents, highlighting a significant level of caffeic acid (4978.00 ± 900.00 mg/100 g dry matter (DM)), trans-cinnamic acid (2144.20 ± 37.60 mg/100 g DM) and quercetin 3-ß-D-glucoside (236.60 ± 3.12 mg/100 g DM). The pectin extraction yield was approximatively 24%, with a degree of esterification of 37.68 ± 1.74%, and a methoxyl content of 5.58 ± 0.88%. The freeze-dried apple pomace was added in a different ratio as a supplement to cultural medium of Loigolactobacillus bifermentans MIUG BL 16, suggesting a significant prebiotic effect (p < 0.05) at concentration between 1% and 2%. The apple pomace was used to design three freeze-dried ingredients containing probiotic, with a high level of polyphenolic content (6.38 ± 0.14 mg gallic acid equivalents/g DM) and antioxidant activity (42.25 ± 4.58 mMol Trolox/g DM) for the powder containing apple pomace ethanolic extract. When inulin was used as a prebiotic adjuvant, the obtained powder showed a 6 log/g DM viable cell count. The ingredients were added to fermented vegetable soy milk-based products, allowing us to improve the polyphenolic content, antioxidant activity and viable cell counts. The approach designed in this study allowed us to obtain ingredients suitable to add value to food, whereas premises to align with the current circular economy premises, by reintegrating the industrial waste as sources of high added value compounds, are also provided.

Tailoring the Functional Potential of Red Beet Purées by Inoculation with Lactic Acid Bacteria and Drying.

This study was focused on a comparative analysis of two drying methods, such as convective and infrared drying, on the red beetroot purées with lactic acid bacteria, as a strategy for tailoring the health benefits of the selected plant. For both varieties, the total betalain contents varied from 13.95 ± 0.14 mg/g dry weight in Beta vulgaris var. cylindra when compared with 11.09 ± 0.03 mg/g dry weight in Beta vulgaris var. vulgaris, whereas significant differences were found in total phenolic and flavonoid contents. Significant drying induced changes were found in selected bioactives, in terms of total betalains, flavonoids, and polyphenols, which influenced the antioxidant activities of the purées, structure, and color parameters. In general, infrared technology was more protective, leading to an increase of 20% in flavonoids content. One logarithmic decrease in cell viability was observed in all powders samples. After the in vitro digestion, the betalains decreased, in both gastric and intestinal simulated juices, with a more pronounced profile in infrared processed purées. Textural and rheological analysis of the dried purées highlighted that the infrared drying is milder compared to the conventional one, allowing us to obtain powders with enhanced functional properties, in terms of bioactives content, cell viability, color, and structural and rheological behavior.

Co-Microencapsulation of Anthocyanins from Black Currant Extract and Lactic Acid Bacteria in Biopolymeric Matrices.

Anthocyanins from black currant extract and lactic acid bacteria were co-microencapsulated using a gastro-intestinal-resistant biocomposite of whey protein isolate, inulin, and chitosan, with an encapsulation efficiency of 95.46% ± 1.30% and 87.38% ± 0.48%, respectively. The applied freeze-drying allowed a dark purple stable powder to be obtained, with a satisfactory content of phytochemicals and 11 log colony forming units (CFU)/g dry weight of powder (DW). Confocal laser microscopy displayed a complex system, with several large formations and smaller aggregates inside, consisting of biologically active compounds, lactic acid bacteria cells, and biopolymers. The powder showed good storage stability, with no significant changes in phytochemicals and viable cells over 3 months. An antioxidant activity of 63.64 ± 0.75 mMol Trolox/g DW and an inhibitory effect on α-amylase and α-glucosidase of 87.10% ± 2.08% and 36.96% ± 3.98%, respectively, highlighted the potential biological activities of the co-microencapsulated powder. Significantly, the in vitro digestibility profile showed remarkable protection in the gastric environment, with controlled release in the intestinal simulated environment. The powder was tested by addition into a complex food matrix (yogurt), and the results showed satisfactory stability of biologically active compounds when stored for 21 d at 4 °C. The obtained results confirm the important role of microencapsulation in ensuring a high degree of protection, thus allowing new approaches in developing food ingredients and nutraceuticals, with enhanced functionalities.

Co-Microencapsulation of Anthocyanins from Cornelian Cherry Fruits and Lactic Acid Bacteria in Biopolymeric Matrices by Freeze-Drying: Evidences on Functional Properties and Applications in Food.

Cornus mas was used in this study as a rich source of health-promoting bioactives. The cornelian cherries were used to extract the polyphenols and anthocyanins. The chromatographic profile of the Cornus mas fruit extract revealed the presence of several anthocyanins, mainly delphinidin, cyanidin and pelargonidin glycosides. The extract was co-microencapsulated with Lactobacillus casei ssp. paracasei in a unique combination of whey protein isolates, inulin and chitosan by freeze-drying, with an encapsulation efficiency of 89.16 ± 1.23% for anthocyanins and 80.33 ± 0.44% for lactic acid bacteria. The pink-red colored powder showed a total anthocyanins content of 19.86 ± 1.18 mg cyanidin-3-glucoside/g dry weight (DW), yielding an antioxidant activity of 54.43 ± 0.73 mMol Trolox/g DW. The viable cells were 9.39 × 109 colony forming units (CFU)/g DW. The confocal microscopy analysis revealed the microencapsulated powder as a complex one, with several large formations containing smaller aggregates, consisting of the lactic acid bacteria cells, the cornelian cherries' bioactive compounds and the biopolymers. The powder was tested for stability over 90 days, showing a decrease of 50% in anthocyanins and 37% in flavonoids content, with no significant changes in antioxidant activity and CFU. The powder showed a significant inhibitory effect against the α-amylase of 89.72 ± 1.35% and of 24.13 ± 0.01% for α-glucosidase. In vitro digestibility studies showed a significant release of anthocyanins in gastric juice, followed by a decrease in intestinal simulated conditions. The functional properties of the powder were tested by addition into a yogurt, highlighting a higher and more stable antioxidant activity at storage when compared to the control.

Valorizations of Sweet Cherries Skins Phytochemicals by Extraction, Microencapsulation and Development of Value-Added Food Products.

Sweet cherries are processed in various ways, leading to significant amounts of underutilized by-products that can potentially be used as a source of bioactive compounds, including antioxidants. The present study focuses on identifying ways to exploit bioactive compounds from sweet cherry skins, namely the extraction, microencapsulation, and functionalizing of some food product to obtain added value. The anthocyanins from skins were extracted and encapsulated in a combination of whey proteins isolate and chitosan by freeze-drying, with an encapsulation efficiency of 77.68 ± 2.57%. The powder showed a satisfactory content in polyphenols, of which anthocyanins content was 14.48 ± 1.17 mg cyanidin 3-glucoside/100 g dry weight (D.W.) and antioxidant activity of 85.37 ± 1.18 µM Trolox/100 g D.W. The powder was morphologically analyzed, revealing the presence of coacervates, ranging in size from 12-54 µm, forming large spheresomes (up to 200 µm). The powder was used as a functional ingredient to develop two value-added food products, namely yoghurt and marshmallows. The powder was tested for its prebiotic effect on L. casei 431® in the yoghurt samples during 21 days at 4 °C, when a decrease in viability was found, up to 6 log CFU·g-1. The anthocyanins and antioxidant activity decreased in yoghurt and increased in marshmallows during storage time. The obtained results support the potential use of extracts from underutilized sources in the development of functional ingredients and value-added food products.

Effect of human isolated probiotic bacteria on preventing Campylobacter jejuni colonization of poultry.

This study was performed in order to determine whether human isolated probiotic bacteria can be effective in reducing Campylobacter jejuni infection of chicken intestinal cells, in vitro, and in decreasing its colonization abilities within the chicken gut. Our results show that the probiotic strains Lactobacillus paracasei J. R, L. rhamnosus 15b, L. lactis Y, and L. lactis FOa had a significant effect on C. jejuni invasion of chicken primary cells, with the strongest inhibitory effect detected when a combination of four was administered. In regard to the in vivo effect, using all four strains in one combination prevented mucus colonization in the duodenum and cecum. Moreover, the pathogen load in the lumen of these two compartments was significantly reduced. When probiotics were introduced during the early growth period, the presence of the pathogen in feces was increased (p>0.05), but when they were given during the last week of growth, there was no significant effect. In conclusion, our data indicate that these four new probiotic strains are able to cause modifications in the chicken intestinal mucosa and can reduce the ability of C. jejuni to invade, in vitro, and to colonize, in vivo. These probiotics are now proven to be effective even when introduced in broiler's feed 7 days before slaughter, which makes them cost-effective for the producers.

Virulence characteristics of five new Campylobacter jejuni chicken isolates.

Campylobacter enteritis has emerged as one of the most common forms of human diarrheal illness. In this study we have investigated the virulence potential of five new C. jejuni chicken isolates (RO14, RO19, RO24, RO29 and RO37) originated from private households in the rural regions of Banat and Transylvania in Romania. Following isolation and in vitro virulence assay, on HCT-8 cells, our results show that all the C. jejuni chicken isolates overcome the virulence abilities of the highly virulent strain C. jejuni 81-176. Motility, an important virulence factor was significantly improved in all the new chicken isolates. The ability to survive to the antimicrobial activity of the human serum, to resist to the violent attack of bile acids and to survive in the presence of synthetic antibiotics was increased in all the chicken isolates. However, these were statistically significant only for isolates RO29 and RO37. In conclusion our study shows, based on invasiveness and motility, and also on the data provided by the serum and bile resistance experiments that all the new chicken isolates are able to infect human cells, in vitro, and could potentially represent a health hazard for humans.

Effect of buckwheat flour and oat bran on growth and cell viability of the probiotic strains Lactobacillus rhamnosus IMC 501®, Lactobacillus paracasei IMC 502® and their combination SYNBIO®, in synbiotic fermented milk.

Fermented foods have a great significance since they provide and preserve large quantities of nutritious foods in a wide diversity of flavors, aromas and texture, which enrich the human diet. Originally fermented milks were developed as a means of preserving nutrients and are the most representatives of the category. The first aim of this study was to screen the effect of buckwheat flour and oat bran as prebiotics on the production of probiotic fiber-enriched fermented milks, by investigating the kinetics of acidification of buckwheat flour- and oat bran-supplemented milk fermented by Lactobacillus rhamnosus IMC 501®, Lactobacillus paracasei IMC 502® and their 1:1 combination named SYNBIO®. The probiotic strains viability, pH and sensory characteristics of the fermented fiber-enriched milk products, stored at 4 °C for 28 days were also monitored. The results showed that supplementation of whole milk with the tested probiotic strains and the two vegetable substrates results in a significant faster lowering of the pH. Also, the stability of L. rhamnosus IMC 501®, L. paracasei IMC 502® and SYNBIO® during storage at 4 °C for 28 days in buckwheat flour- and oat bran-supplemented samples was remarkably enhanced. The second aim of the study was to develop a new synbiotic product using the best combination of probiotics and prebiotics by promoting better growth and survival and be acceptable to the consumers with high concentration of probiotic strain. This new product was used to conduct a human feeding trial to validate the fermented milk as a carrier for transporting bacterial cells into the human gastrointestinal tract. The probiotic strains were recovered from fecal samples in 40 out of 40 volunteers fed for 4 weeks one portion per day of synbiotic fermented milk carrying about 10(9) viable cells.