Phytochemical Characterization and Antibacterial Activity of Albanian Juniperus communis and Juniperus oxycedrus Berries and Needle Leaves Extracts.

This paper aims to investigate the phytochemical profile and in vitro antibacterial activity of two juniper species collected in Albania, Juniperus communis and Juniperus oxycedrus. The berries and the needle leaves were subjected to solid-liquid solvent ultrasound-assisted extraction. The phytochemical characterization of the extracts was performed by spectrophotometric and chromatographic means. The extract of J. communis berries (JcB) showed a higher total phenolic and flavonoid content (3.04 ± 0.09 mg GAE/g DW, and 1.14 ± 0.36 mg QE/g DW, respectively), also a higher antioxidant activity from DPPH and ABTS radical screening assays, compared to J. oxycedrus berries (JoxB) extract. The extract of J. oxycedrus needle leaves (JoxL) prevailed in total flavonoid content (10.55 ± 0.24 mg QE/g DW), and ABTS assays (1.83 ± 0.01 mM TE/g DW), compared to the extract of J. communis needle leaves (JcL). The chromatographic analysis revealed the presence of ellagic acid and kaempferol in all the samples. Ellagic acid was the main identified compound with the highest quantity in the extracts of JoxB, JoxL, and JcB with an average of 445.69 ± 0.96 µg/g, 2890.05 ± 0.29 µg/g, and 8133.83 ± 4.03 µg/g, respectively. The antibacterial potential of the ethanolic extracts was evaluated on Bacillus spp., Escherichia coli, and Staphylococcus aureus. In the Agar Well Diffusion Assay, it was observed that all the tested bacterial strains were sensitive to the extracts, whereas selected extracts showed a similar inhibition activity rate compared with the antibiotic substance (Chloramphenicol), used as a positive control. The extracts showed a similar minimal inhibitory and bactericidal concentration for the individual bacterial strains, suggesting that J. communis and J. oxycedrus extracts have a similar potential in antibacterial activity.

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.

Advanced interactional characterization of the inhibitory effect of anthocyanin extract from Hibiscus sabdariffa L. on apple polyphenol oxidase.

In this study, a comprehensive approach to advance the inhibitory effect of Hibiscus sabdariffa extract on apple polyphenol oxidase (PPO) was performed. PPO was extracted, purified, and characterized for optimal activity, whereas response surface methodology generated a quadratic polynomial model to fit the experimental results for hibiscus extraction. The optimum conditions allowed to predict a maximum recovery of anthocyanins (256.11 mg delphinidin-3-O-glucoside/g), with a validated value of 272.87 mg delphinidin-3-O-glucoside/g dry weight (DW). The chromatographic methods highlighted the presence of gallic acid (36,812.90 µg/g DW extract), myricetin (141,933.84 µg/g DW extract), caffeic acid (101,394.07 µg/g DW extract), sinapic acid (1157.46 µg/g DW extract), kaempferol (2136.76 µg/g DW extract), and delphinidin 3-O-ß-d-glucoside (226,367.08 µg/g DW extract). The inactivation of PPO followed a first-order kinetic model. A temperature-mediated flexible fit between PPO and anthocyanins was suggested, whereas the molecular docking tests indicated that PPO is a good receptor for cafestol, gallic acid, and catechin, involving hydrophobic and hydrogen bond interactions. PRACTICAL APPLICATION: It is well known that enzymatic browning is one of the most important challenges in the industrial minimal processing of selected fruit and vegetable products. Novel inhibitors for polyphenol oxidase are proposed in this study by using an anthocyanin-enriched extract from Hibiscus sabdariffa L. Based on our results, combining the chemical effect of phytochemicals from hibiscus extract with different functional groups with minimal heating could be an interesting approach for the development of a new strategy to inhibit apple polyphenol oxidase.

Technologically Driven Approaches for the Integrative Use of Wild Blackthorn (Prunus spinosa L.) Fruits in Foods and Nutraceuticals.

Different technological approaches were used in this study for the valorization of blackthorn (Prunus spinosa L.) fruits in marmalade, jam, jelly, and nutraceuticals. Marmalade showed the highest concentrations of polyphenols (7.61 ± 0.05 mg gallic acid equivalents/g dry weight (DW)) and flavonoids (4.93 ± 0.22 mg catechin equivalents/g DW), whereas jam retained the highest content of anthocyanins (66.87 ± 1.18 mg cyanidin-3-O-glucoside equivalents/g DW). A good correlation between polyphenol and flavonoid contents and antioxidant activity was found, the highest value being 21.29 ± 1.36 mmol Trolox/g DW for marmalade. Alternatively, the fresh pulp was enriched with inulin, followed by inoculation with Lactobacillus acidophilus, and freeze-dried, allowing a powder to be obtained with a viable cell content of 6.27 × 107 CFU/g DW. A chromatographic analysis of blackthorn skin revealed that myricetin (2.04 ± 0.04 mg/g DW) was the main flavonoid, followed by (+)-catechin (1.80 ± 0.08 mg/g DW), (-)-epicatechin (0.96 ± 0.02 mg/g DW), and vanillic acid (0.94 ± 0.09 mg/g DW). The representative anthocyanins were cyanidin 3-O-glucoside, cyanidin 3-O-rutinoside, and peonidin 3-O-glucoside, with an average concentration of 0.75 mg/g DW. The skin extract showed comparable IC50 values for tyrosinase (1.72 ± 0.12 mg/mL), α-amylase (1.17 ± 0.13 mg/mL), and α-glucosidase (1.25 ± 0.26 mg/mL). The possible use of kernels as calorific agents was demonstrated through the evaluation of calorific power of 4.9 kWh/kg.

Kombucha and Water Kefir Grains Microbiomes' Symbiotic Contribution to Postbiotics Enhancement.

Wild artisanal cultures, such as a symbiotic culture of bacteria and yeasts (SCOBY) and water kefir grains (WKG), represent a complex microorganism consortia that is composed of yeasts and lactic and acetic acid bacteria, with large strains of diversity and abundance. The fermented products (FPs) obtained by the microbiome's contribution can be included in functional products due to their meta-biotics (pre-, pro-, post-, and paraprobiotics) as a result of complex and synergistic associations as well as due to the metabolic functionality. In this study, consortia of both SCOBY and WKG were involved in the co-fermentation of a newly formulated substrate that was further analysed, aiming at increasing the postbiotic composition of the FPs. Plackett-Burman (PBD) and Response Surface Methodology (RSM) techniques were employed for the experimental designs to select and optimise several parameters that have an influence on the lyophilised starter cultures of SCOBY and WKG activity as a multiple inoculum. Tea concentration (1-3%), sugar concentration (5-10%), raisins concentration (3-6%), SCOBY lyophilised culture concentration (0.2-0.5%), WKG lyophilised culture concentration (0.2-0.5%), and fermentation time (5-7 days) were considered the independent variables for mathematical analysis and fermentation conditions' optimisation. Antimicrobial activity against Bacillus subtilis MIUG B1, Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922, and Aspergillus niger MIUG M5, antioxidant capacity (DPPH), pH and the total acidity (TA) were evaluated as responses. The rich postbiotic bioactive composition of the FP obtained in optimised biotechnological conditions highlighted the usefulness of the artisanal co-cultures, through their symbiotic metabolic interactions for the improvement of bioactive potential.

Customized Technological Designs to Improve the Traditional Use of Rosa canina Fruits in Foods and Ingredients.

The rosehip fruits from Romanian spontaneous flora were valorized in three different customized variants, including jellified products, juices, and a nutraceutical. Prior to the technological design, the rosehip samples were characterized for individual polyphenolic content. The samples (non)-enzymatically treated showed epicatechin as the major compound, whereas the enzymatic treatment enhanced the extraction of quercetin (40.23 ± 0.48 mg/100 g DW extract), gallic acid (9.74 ± 0.24 mg/100 g DW extract), and kaempferol. Different ratios and types of sugars were added to obtain jellified products, leading to a significantly different bioactive profile. The sugar-free and steviol samples showed the highest content in carotenoids (89.36 ± 0.06 mg/g dry weight (DW) and 39.22 ± 1.16 mg/g DW, respectively), leading to the highest antioxidant activity (8.19 ± 0.80 mMol Trolox/g DW and 20.16 ± 0.31 mMol Trolox/g DW, respectively). The gelling capacity increased with the increase in sugar content. The enzymatically treated rosehip fruit pulp was mixed in various ratios with apple juice, followed by pasteurization. The polyphenolic content was approximately two times higher in the blended juices (20.75 ± 1.40 mg gallic acid equivalents (GAE)/100 mL). The overall impression highlighted the preferences of panelists for sugar-free products, whereas adding apple juice significantly influenced their preferences. The fresh pulp was enhanced with pectin, followed by inoculation with Lactobacillus acidophilus, and freeze-dried showed satisfactory cell viability (approximately 7 log CFU/g DW), whereas an antidiabetic, anti-tyrosinase and anti-obesity potential of the powder was suggested. Our results provide enough evidence for customized processing of rosehip fruits in foods and nutraceuticals as a response to consumers' choices, highlighting the bioactive compounds and nutrient contents, whereas selected in vitro health-related evidence was suggested.

CO2 supercritical extraction and microencapsulation of oleoresins from rosehip fruits for getting powders with multiple applications.

The supercritical fluids extraction (SFE) was used to extract the oleoresins from rosehip, followed by an in-depth phytochemical analysis and the development of two design-customized powders for different food and pharmaceutical applications. The SFE experiments allowed obtaining an oleoresins extraction yield of 11.85%. Two fractions were separated (S40 and S45), with significantly different phytochemical profile (p < 0.05), highlighting the efficiency of extraction of fatty acids in S40 extract, whereas the extraction of polyphenols, phytosterols, carotenoids and polyphenols was favored in S45 extract. The phytochemical profile revealed that the linoleic acid (C18:2) and α-linolenic acid (C18:3) represented approximatively 82% and 58% from the total fatty acid content in S40 and S45, respectively. α-Tocopherol and γ-tocopherol prevailed in both extract fractions, with a higher concentration in S45 (229.66 mg/g dry matter (DM) and 112.36 mg/g DM, respectively), whereas ß-sitosterol was the major phytosterol in S45 fraction (118.75 mg/g DM). The S40 fraction was used to design two microencapsulated powders, by combining emulsification, complex coarcevation and freeze-drying. In order to develop new wall materials, with unique properties, the soy protein isolates were used for cross-linked reactions, by using an approach in one step (transglutaminase mediated) (coded as N) and two-steps (heat-induced and transglutaminase mediated) (coded as T). The N powder showed a better phytochemical content, leading to a higher antioxidant activity (5.27 mM Trolox equivalents/g DM), whereas for variant T, the bioactive were apparently doubled encapsulated.

Valorization of Bioactive Compounds from Two Underutilized Wild Fruits by Microencapsulation in Order to Formulate Value-Added Food Products.

Currently, microencapsulation has become a viable method of nutrient delivery for the food industry. This work microencapsulated the bioactive compounds extracted from two neglected species (Balanites aegyptiaca and Ziziphus mauritiana) by freeze-drying. A combination of wall materials (whey protein and pectin; soy protein and maltodextrin) was chosen to prepare the microcapsules. The phytochemical and physicochemical characterization of the microcapsules was then carried out. The encapsulation yield ranged from 82.77% to 96.05% for Balanites and Ziziphus, respectively, whereas the efficiency was 76.18 ± 1.39% and 80.93 ± 1.71%. The stimulated in vitro gastrointestinal test showed that encapsulation increased the bioavailability of the bioactive compounds. The total carotenoids were the most bioavailable compounds with 85.89 ± 0.06% for Ziziphus and 70.46 ± 1.10% for Balanites, followed by total flavonoids for Zizyphus with 63.27 ± 1.56%. Furthermore, regardless of species or wavelengths, the HPLC analysis resulted in the identification of 17 bioactive metabolites. The predominant one was epicatechin, whose level ranged from 231.52 ± 5.06 to 250.99 ± 3.72 mg/100 g DW in Ziziphus and 91.80 ± 3.85 to 116.40 ± 4.09 mg/100 g DW in Balanites. In estimating the enzyme inhibition and antioxidant power, both studied fruits showed antidiabetic, inflammatory, and antioxidant effects. These findings suggest that natural bioactive compounds are abundant in the fruits of Z. mauritiana and B. aegyptiaca and could be a valuable source for the food and pharmaceutical industries.

Optimizing of the extraction conditions for anthocyanin's from purple corn flour (Zea mays L): Evidences on selected properties of optimized extract.

In this study, the Response Surface Methodology was used to optimize the extraction conditions for anthocyanins from purple corn flour (PCF), whereas the optimized extract was characterized in terms of phytochemicals content, inhibitory activity against metabolic associated enzymes and the potential to stimulate the metabolic activity of yeasts. The optimized conditions using a Box-Behnken design (BBD) of response surface methodology (RSM) were as follow: temperature (39 °C), extraction time (5 h), liquid/solid ratio (30 mL/g) and ethanol concentration (73 %), yielding a maximum response predicted and experimental values of 13.77 and 14.04 ± 0.02 mg cyanidin 3-O-glucoside equivalents (C3G)/g dry weight (DW). The extract showed a high content in myricetin, quercetin 3-ß-d-glucoside, kaempferol, whereas the main anthocyanin was C3G. The extract was test the effect on metabolic activity of Saccharomyces cerevisiae, with the highest multiplication rate obtained at 1.2 mg/100 mL. The extract showed potential antidiabetic, hypocholesterolemic and preventive effects against Parkinson's disease and melanoma. The findings provide the optimum conditions to obtain an anthocyanin enriched extract, for multiple applications.

Tailoring the Optimized Fermentation Conditions of SCOBY-Based Membranes and Milk Kefir Grains to Promote Various Functional Properties.

Kombucha culture (named SCOBY-Symbiotic Culture of Bacteria and Yeasts) and milk kefir grains represent multiple consortia of wild microorganisms that include lactic acid bacteria, acetic acid bacteria and yeasts with valuable functional properties. Their fermentative potential provides a wide range of derivate metabiotics (prebiotics, probiotics, postbiotics and paraprobiotics) with valuable in vitro and in vivo benefits. This study targeted the evaluation of the functionality of a co-culture of SCOBY-based membranes and milk kefir grains, used as freeze-dried starter cultures, for the fermentation of a newly formulated medium based on black tea infusion, supplemented with bovine colostrum and sugar, in order to produce bioactive compounds with functional properties. The design and optimization of the biotechnological process were achieved by using the Plackett-Burman experimental design (six factorial points, three center points) and the response surface methodology and central composite design (three factorial points, six axial points and two center points in axial) tools. The statistical analysis and the mathematical modelling of the responses such as the pH, titratable acidity, antioxidant activity and antimicrobial activity (against Bacillus subtilis, Escherichia coli, Staphylococcus aureus and Aspergillus niger) were investigated. Further, the composition of organic acids, polyphenols and flavonoids of the fermented product obtained under the optimized fermentation conditions was also analyzed. The fermentation of the medium containing 6.27% (w/v) bovine colostrum powder, 1.64% (w/v) black tea, 7.5% (w/w) sugar, pH 6.7, with an inoculum based of 0.36% (w/v) milk kefir grains powder and 0.5% (w/v) SCOBY-based membrane (both as freeze-dried culture), at 30 °C, for 5 days, in an aerobic stationary system, revealed an antifungal activity between 80 and 100% against Aspergillus niger, an antibacterial activity of 8-22 mm against Escherichia coli and Bacillus spp. And a titratable acidity of 445 °Th. The chemical composition of the obtained product had a positive impact on the functional properties of the fermented products in terms of the antimicrobial and antioxidant properties.

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.

Novel Insights for Metabiotics Production by Using Artisanal Probiotic Cultures.

Wild probiotic consortia of microorganisms (bacteria and yeasts) associated in the artisanal cultures' microbiota (milk kefir grains, water kefir grains and kombucha) are considered valuable promoters for metabiotics (prebiotics, probiotics, postbiotics and paraprobiotics) production. The beneficial effects of the fermented products obtained with the artisanal cultures on human well-being are described by centuries and the interest for them is continuously increasing. The wild origin and microbial diversity of these above-mentioned consortia give them extraordinary protection capacity against microbiological contaminants in unusual physico-chemical conditions and unique fermentative behaviour. This review summarizes the state of the art for the wild artisanal cultures (milk and water kefir grains, respectively, kombucha-SCOBY), their symbiotic functionality, and the ability to ferment unconventional substrates in order to obtain valuable bioactive compounds with in vitro and in vivo beneficial functional properties. Due to the necessity of the bioactives production and their use as metabiotics in the modern consumer's life, artisanal cultures are the perfect sources able to biosynthesize complex functional metabolites (bioactive peptides, antimicrobials, polysaccharides, enzymes, vitamins, cell wall components). Depending on the purposes of the biotechnological fermentation processes, artisanal cultures can be used as starters on different substrates. Current studies show that the microbial synergy between bacteria-yeast and/or bacteria-offers new perspectives to develop functional products (food, feeds, and ingredients) with a great impact on life quality.

Selection of Wild Lactic Acid Bacteria Strains as Promoters of Postbiotics in Gluten-Free Sourdoughs.

The occurrence of inflammatory responses in humans is frequently associated with food intolerances and is likely to give rise to irritable bowel disease. The use of conventional or unconventional flours to produce gluten-free baking doughs brings important technological and nutritional challenges, and the use of the sourdough biotechnology has the potential to overcome such limitations. In addition, the typical metabolic transformations carried out by Lactic Acid Bacteria (LAB) can become an important biotechnological process for the nutritional fortification and functionalization of sourdoughs due to the resulting postbiotics. In such a context, this research work aimed at isolating and selecting new LAB strains that resort to a wide range of natural environments and food matrices to be ultimately employed as starter cultures in gluten-free sourdough fermentations. Nineteen LAB strains belonging to the genera of Lactobacillus, Leuconostoc, Pediococcus, and Streptococcus were isolated, and the selection criteria encompassed their acidification capacity in fermentations carried out on chickpea, quinoa, and buckwheat flour extracts; the capacity to produce exopolysaccharides (EPS); and the antimicrobial activity against food spoilage molds and bacteria. Moreover, the stability of the LAB metabolites after the fermentation of the gluten-free flour extracts submitted to thermal and acidic treatments was also assessed.