Chemical Profile Characterization of Fruit and Vegetable Juices after Fermentation with Probiotic Strains.

Lactic acid bacteria (LAB) are widely applied for fermentation purposes in dairy and non-dairy food matrices with beneficial technological and health-promoting properties. This study describes the effect of two lactic acid bacteria, namely, Lactiplantibacillus paracasei SP5 and Pediococcus pentosaceus SP2, on the phenolic profiles, antioxidant activities, total phenolic content (TPC), carotenoid content, and sensorial profile of two different mixed fruit juices. After 48 h of fermentation, both LABs retained viability over 9 Log CFU/mL in both juices. The TPC, zeaxanthin + lutein, ß-carotene content, and antioxidant activity (AA) were elevated for both LABs and mixed juices after 48 h of fermentation compared to control samples. Regarding the phenolic profile, both juices exhibited a significant decrease in chlorogenic acid levels, while quinic acid and tyrosol concentrations showed notable increases.

Case Studies of Small-Medium Food Enterprises around the World: Major Constraints and Benefits from the Implementation of Food Safety Management Systems.

Global food safety and security are key principles to be followed in the context of the implementation of food safety management systems. The objective of this paper is to assess the contemporary developments of Food Safety Management System standards (FSMS) worldwide and to identify the primary constraints and advantages associated with their implementation by small and medium-sized enterprises across different regions. The effectiveness of these systems has also been evaluated. 116 case studies have been employed across developing and developed regions worldwide across 27 primary food sectors. After the implementation of FSMS, there was a significant increase in the percentage of companies that have implemented the international FSMS, both in developed (16.7% to 63.9%) and developing countries (26.6% to 48.1%). Certification has also increased from 34.2% to 59.6% in the total sample, namely from 33.3% to 61.1% in developed countries and from 34.6% to 59.0% in developing countries. There was a significant increase in medium vs. small company size (57.1% to 62.3%, p = 0.046), only in developing countries. Food safety culture and manager leadership implementation were increased to over 80% after FSMS implementation in both developed and developing countries (p < 0.001). Training, resources, and technology adequacy were also increased in all companies (p < 0.001).

Nutritional Improvements of Sourdough Breads Made with Freeze-Dried Functional Adjuncts Based on Probiotic Lactiplantibacillus plantarum subsp. plantarum and Pomegranate Juice.

New types of sourdough breads are proposed, made with freeze-dried sourdough adjuncts based on: (i) Lactiplantibacillus plantarum subsp. plantarum ATCC 14917, a potential probiotic (LP) alone or (ii) with the addition of unfermented pomegranate juice (LPPO) and (iii) pomegranate juice fermented by the same strain (POLP). Physicochemical, microbiological, and nutritional characteristics (in vitro antioxidant capacity, AC, total phenolics, TPC, and phytate content) of the breads were evaluated and compared with commercial sourdough bread. All adjuncts performed well; the best results being those obtained by POLP. Specifically, the highest acidity (9.95 mL of 0.1 M NaOH) and organic acid content (3.02 and 0.95 g/kg, lactic and acetic acid, respectively) as well as better resistance to mold and rope spoilage (12 and 13 days, respectively) were observed for POLP3 bread (sourdough with 6% POLP). Significant nutritional improvements were observed by all adjuncts, in terms of TPC, AC, and phytate reduction (103 mg gallic acid/100 g, 232 mg Trolox/100 g, and 90.2%, respectively, for POLP3). In all cases, the higher the amount of adjunct, the better the results. Finally, the good sensory properties of the products indicate the suitability of the proposed adjuncts for sourdough breadmaking, while their application in freeze-dried, powdered form can facilitate commercial application.

Microbiological stability and sensorial valorization of thyme and oregano essential oils alone or combined with ethanolic pomegranate extracts in wine marinated pork meat.

In the present study, wine-based marinades containing ethanolic extract from pomegranate (Punica granatum L.), alone or in combination with two Essential Oils (Thyme & Oregano), were used for pork fillets marination and their antimicrobial activity, as well as their sensorial impact were evaluated. Likewise, the marinades exhibited promising results concerning their recorded antimicrobial activity versus Enterobacteriaceae, Total Mesophilic Bacteria, Yeasts/molds, Staphylococcus spp., Pseudomonas spp. & Lactic Acid Bacteria (LAB). The outcome demonstrated that pork fillets marinated with wine containing ethanolic extract of pomegranate and Oregano Essential Oil were more resistant to spoilage compared to all other samples; thus, their shelf-life was significantly extended (4 days in some cases). Triterpenes (maslinic, oleanolic and betulinic acid), monoterpenes (p-cymene, carvacrol, thymol, limonene), organic acids (citric & malic acid) and phenols, were the main constituents found in the plant extract, the wine and Essential Oils applied, as determined through LC-QTOF/MS and HPLC analysis. Additionally, the sensorial properties (color, tenderness, flavor and juiciness) of the marinated meat samples were not negatively influenced. Consequently, marinades of this type could be used as natural preservatives in meat products, with satisfying antimicrobial and organoleptic results.

The In Vitro Assessment of Antibacterial and Antioxidant Efficacy in Rosa damascena and Hypericum perforatum Extracts against Pathogenic Strains in the Interplay of Dental Caries, Oral Health, and Food Microbiota.

The rising demand for novel antibiotic agents prompts an investigation into natural resources, notably plant-derived compounds. In this study, various extracts (aqueous, ethanolic, aqueous-ethanolic, and enzymatic) of Rosa damascena and Hypericum perforatum were systematically evaluated against bacterial strains isolated from dental lesions (n = 6) and food sources (raw milk and broiler carcass, n = 2). Minimal inhibitory concentration (MIC), minimal bactericidal concentration (MBC), antibiofilm activity, and time-kill kinetics were assessed across a range of extract concentrations, revealing a dose-responsive effect. Notably, some extracts exhibited superior antibacterial efficacy compared to standard clinical antibiotics, and the time-kill kinetics demonstrated a rapid elimination of bacterial loads within 24 h. The susceptibility pattern proved strain-specific, contingent upon the extract type, yet all tested pathogens exhibited sensitivity. The identified extracts, rich in phenolic and polyphenolic compounds, as well as other antioxidant properties, contributed to their remarkable antibiotic effects. This comprehensive investigation not only highlights the potential of Rosa damascena and Hypericum perforatum extracts as potent antibacterial agents against diverse bacterial strains including caries pathogens, but also underscores their rapid action and dose-dependent efficacy. The findings suggest a promising avenue for harnessing plant-derived compounds in the development of novel antimicrobial strategies against dental caries and other oral inflammations, bridging the gap between natural resources and antibiotic discovery.

Genomic Insight Into Lacticaseibacillus paracasei SP5, Reveals Genes and Gene Clusters of Probiotic Interest and Biotechnological Potential.

The Lacticaseibacillus paracasei species is comprised by nomadic bacteria inhabiting a wide variety of ecological niches, from fermented foodstuffs to host-associated microenvironments. Lc. paracasei SP5 is a novel strain, originally isolated from kefir grains that presents desirable probiotic and biotechnological attributes. In this study, we applied genomic tools to further characterize the probiotic and biotechnological potential of the strain. Firstly, whole genome sequencing and assembly, were performed to construct the chromosome map of the strain and determine its genomic stability. Lc. paracasei SP5 carriers several insertion sequences, however, no plasmids or mobile elements were detected. Furthermore, phylogenomic and comparative genomic analyses were utilized to study the nomadic attributes of the strain, and more specifically, its metabolic capacity and ability to withstand environmental stresses imposed during food processing and passage through the gastrointestinal (GI) tract. More specifically, Kyoto Encyclopedia of Genes and Genomes (KEGG) and Carbohydrate-active enzyme (CAZymes) analyses provided evidence for the ability of the stain to utilize an array of carbohydrates as growth substrates. Consequently, genes for heat, cold, osmotic shock, acidic pH, and bile salt tolerance were annotated. Importantly bioinformatic analysis showed that the novel strain does not harbor acquired antimicrobial resistance genes nor virulence factors, in agreement with previous experimental data. Putative bacteriocin biosynthesis clusters were identified using BAGEL4, suggesting its potential antimicrobial activity. Concerning microbe-host interactions, adhesins, moonlighting proteins, exopolysaccharide (EPS) biosynthesis genes and pilins mediating the adhesive phenotype were, also, pinpointed in the genome of Lc. paracasei SP5. Validation of this phenotype was performed by employing a microbiological method and confocal microscopy. Conclusively, Lc. paracasei SP5 harbors genes necessary for the manifestation of the probiotic character and application in the food industry. Upcoming studies will focus on the mechanisms of action of the novel strain at multiple levels.

Plant Extract and Essential Oil Application against Food-Borne Pathogens in Raw Pork Meat.

Herbal and plant extracts are being applied for a wide range of foods against different types of food-borne pathogens. In the present study, ethanolic and aqueous extracts (2% w/v) from cranberry (Vaccinium macrocarpon) and pomegranate (Punica granatum L.) plants were applied alone or in combination with two essential oils (thyme and oregano in a concentration of 0.150 µg/g) in pork meatballs and their antimicrobial activity was estimated. The extracts exhibited promising results (aqueous and ethanolic extracts of pomegranate and cranberry in a food-compatible concentration of 2% w/v) were applied to raw pork meatball production and their antimicrobial activity was recorded versus Enterobacteriaceae, total mesophilic bacteria, yeasts/molds, Staphylococcus spp., Pseudomonas spp. and lactic acid bacteria (LAB). The outcome demonstrated that meatballs containing aqueous extracts of pomegranate were more resistant to spoilage compared to all the other samples since they were preserved for more days. The chemical profiles of plant extracts were determined through LC-QTOF/MS and the chemical composition of the essential oils applied was determined with the use of GC/MS in order to identify the substances involved in the observed antimicrobial activity. Phenolic acids (quinic acid, chlorogenic acid), monoterpenes (p-cymene, carvacrol, thymol, limonene), organic acids (citric acid) and phenols were the main constituents found in the plant extracts and essential oils applied. These extracts of plant origin could be used as natural preservatives in meat products, even in low concentrations.

Safety and Efficacy of RAD001 (Everolimus) Administered Upon Relapse During or After Adjuvant Treatment in Post-menopausal Women With Hormone Receptor Positive, HER2/neu Negative Locally Advanced or Metastatic Breast Cancer (CRAD001JGR08 "MELPOMENI" study).

BACKGROUND/AIM: This study aimed to provide real-world safety and effectiveness data of everolimus (EVE) plus exemestane (EXE) in estrogen receptor positive/human epidermal growth factor receptor 2 negative (ER+/HER2-) advanced breast cancer (aBC). PATIENTS AND METHODS: This prospective observational study was conducted by 19 hospital-based oncologists in Greece. Eligible patients were treated with EVE+EXE in the first-line setting; EVE was initiated according to the approved label. RESULTS: Overall, 75 eligible patients (mean age: 66.9 years; visceral metastases: 49.3%; bone-only metastases: 37.3%) were included in the effectiveness analyses. Over a median (interquartile range) of 12.1 months (range=4.2-20.5 months) of EVE treatment, the median progression-free survival was 18.0 months and the overall response rate was 22.7%. Among patients that received ≥1 EVE dose (n=80), the incidence of EVE-related adverse events was 72.5% (serious: 55.0%); stomatitis (22.5%), fatigue (22.5%), pneumonitis (18.8%); and cough (18.8%) were the most common. CONCLUSION: In the routine care in Greece, EVE demonstrates clinical benefit and a predictable safety profile.

mRNA expression of specific HER ligands and their association with clinical outcome in patients with metastatic breast cancer treated with trastuzumab.

Prognostic and predictive biomarkers are being studied for the diagnosis and treatment of breast cancer. The present study retrospectively assessed the mRNA expression of HER family receptor ligands and of other potential prognostic biomarkers and their association with time to progression (TTP), survival and clinicopathological characteristics in patients with metastatic breast cancer (MBC) treated with trastuzumab. A total of 145 tumour tissue samples were analysed. mRNA expression analysis of the transcripts of interest was performed and the association of these markers with selected clinicopathological parameters was examined. HER2 status was centrally re-evaluated. Only 67.6% of patients were truly HER2-positive according to the central HER2 re-evaluation. Heparin binding epidermal growth factor (EGF)-like growth factor, transforming growth factor ß1 (TGFB1) and thyroid hormone receptor α (THRA) mRNA expression was higher in HER2-positive patients (P=0.026, P<0.001 and P<0.001). Insulin-like growth factor binding protein 4 was correlated with retinoic acid receptor α, TGFB1 and THRA (rho=0.45, rho=0.60 and rho=0.45). In HER2-positive patients, high neuregulin 1 and high betacellulin were unfavourable factors for TTP [hazard ratio (HR) = 1.78, P=0.040 and HR=2.00, P=0.043, respectively]. In patients with de novo MBC, high EGF expression was associated with a non-significant prolongation of TTP (HR=0.52, P=0.080) and significantly longer survival (HR=0.40, P=0.020). The present study examined clinical and biological implications of specific genes and it was concluded that their expression has an impact on the outcome of trastuzumab-treated patients with MBC.

In Vitro Screening Potential Antibacterial Properties of the Greek Oregano Honey against Clinical Isolates of Helicobacter pylori.

Oregano honey is an exceedingly rare and distinct product, not commercially available, produced by bees bred in oregano fields of alpine altitudes at the mountainous area of Epirus, Greece. In ethnic popular medicine, this product is used as a therapeutic in various gastric diseases. To test this hypothesis, 14 strains of Helicobacter pylori (H. pylori), 6 isolated from gastric ulcers and 8 from cases of clinical gastritis, were employed in the present study. The above bacterial strains were exposed to various concentrations (75% v/v, 50% v/v, 25% v/v, 12.5% v/v, and 6% v/v) of 50 oregano honey samples by using the agar well method and the inhibition zones observed around each well were recorded. Although the inhibitory zones of the H. pylori isolated from the gastric ulcers were wide enough (0-34 mm), those strains, in general, appeared more resistant than the other eight (0-58 mm). The same result was observed when the same strains were tested against six antibiotics used in clinical practice. Extracts of oregano honey were prepared by extraction with four different organic solvents. N-hexane and chloroform extracts had the most potent antibacterial action. Finally, pure oregano honey and diethyl ether extracts of honey showed significant inhibitory activity against urease secreted by the pathogen. These results strongly indicate the susceptibility of H. pylori strains to the oregano honey by more than one mode of action. Consequently, this variety of honey seems to have potential therapeutic properties against gastric ulcers and gastritis, thus explaining the preference of the locals towards this traditional remedy.

Microbiological and Chemical Properties of Chokeberry Juice Fermented by Novel Lactic Acid Bacteria with Potential Probiotic Properties during Fermentation at 4 °C for 4 Weeks.

On the frame of this research survey, a novel potentially probiotic strain (Lactobacillus paracasei SP5) recently isolated from kefir grains was evaluated for chokeberry juice fermentation. Chokeberry juice was retrieved from the variety Aronia melanocarpa, a plant known to provide small, dark berries and to be one of the richest sources of antioxidants. The juice was subsequently fermented inoculating L. paracasei SP5 for 48 h at 30 °C. The fermented juices were left at 4 °C and tested regarding microbiological and physicochemical characteristics for 4 weeks. The potentially probiotic strain was proved capable of performing lactic acid fermentation at 30 °C. Cell viability of L. paracasei was detected in high levels during fermentation and the whole storage period, while the fermented juice showed higher levels of viability in juice with 40.3 g/L of initial sugar concentration. No ethanol was detected in the final fermented juice. Fermented chokeberry juice was characterized by aromatic desirable volatiles, which were retained in adequate levels for the whole storage period. Specifically, the occurrence of organic esters detected in fermented juices is considered as positive evidence of the provision of fruity and floral notes to the final product. During storage, total phenolics content and antioxidant activity were observed in higher levels in fermented chokeberry juice compared with non-fermented juice. Subsequently, fermentation of chokeberry juice by potentially probiotic lactic acid bacteria could provide high industrialization potential, providing the market with a nutritional beverage of good volatile quality with an enhanced shelf-life compared with an unfermented fresh juice.

Assessment of Antimicrobial Activity of Pomegranate, Cranberry, and Black Chokeberry Extracts against Foodborne Pathogens.

Herbal and plant extracts are being applied to a wide range of foods against different types of foodborne pathogens. In the present study, ethanolic and aqueous extracts of different concentrations (5% v/v, 10% v/v, and 20% v/v) from cranberry (Vaccinium macrocarpon), black chokeberry (Aronia melanocarpa), and pomegranate (Punica granatum L.) plants were applied in five concentrations (62.5 to 1000 mg/mL) against foodborne strains of Staphylococcus aureus,Escherichia coli, and Streptococcus pyogenes. The results revealed that a low concentration of solvents (5% v/v) did not exhibit decreased antimicrobial activity in comparison with higher solvent concentrations (10% and 20% v/v). Additionally, both aqueous and ethanolic extracts were highly effective against pathogens even in their low concentrations (62.5 mg/mL and 125 mg/mL). Likewise, the extracts exhibited promising results (aqueous extracts of pomegranate, cranberry, and black chokeberry in a food-compatible concentration of 2% w/v) were applied to raw pork meatball production, and their antimicrobial activity was recorded versus Enterobacteriaceae, total mesophilic bacteria (TMB), lactic acid bacteria (LAB), Staphylococcus spp., Pseudomonas spp., and yeasts/molds. The outcome demonstrated that meatballs that contained aqueous extracts of pomegranate were more resistant to spoilage compared to all of the other samples, as it was preserved for more days. Likewise, these extracts of a plant origin could be used as natural preservatives in meat products, even in their low concentrations.

Isolation of a Lactobacillus paracasei Strain with Probiotic Attributes from Kefir Grains.

Κefir is a rich source of potentially probiotic bacteria. In the present study, firstly, in vitro screening for probiotic characteristics of ten lactic acid bacteria (LAB) isolated from kefir grains was performed. Strain AGR 4 was selected for further studies. Molecular characterization of strain AGR 4, confirmed that AGR 4 belongs to the Lactobacillus paracasei (reclassified to Lacticaseibacillus paracasei subsp. paracasei) species. Further testing revealed that L. paracasei AGR 4 displayed adhesion capacity on human adenocarcinoma cells, HT-29, similar to that of the reference strain, L. casei ATCC 393. In addition, the novel strain exerted significant time- and dose-dependent antiproliferative activity against HT-29 cells and human melanoma cell line, A375, as demonstrated by the sulforhodamine B cytotoxicity assay. Flow cytometry analysis was employed to investigate the mechanism of cellular death; however, it was found that AGR 4 did not act by inducing cell cycle arrest and/or apoptotic cell death. Taken together, these findings promote the probiotic character of the newly isolated strain L. paracasei AGR 4, while further studies are needed for the detailed description of its biological properties.

Assessment of the Immunomodulatory Properties of the Probiotic Strain Lactobacillus paracasei K5 in vitro and In Vivo.

Lactobacillus paracasei K5 is a lactic acid bacteria (LAB) strain that has been isolated from dairy products. Previous studies have established its probiotic potential in a series of in vitro tests, including molecular characterization, safety profiling, and tolerability of the gastrointestinal tract conditions. To characterize its beneficial actions on the host, we have shown previously that L. paracasei K5 adheres to Caco-2 cells and exerts anti-proliferative effects through the induction of apoptosis. In the present study, we focused on the immunomodulatory potential of this strain. We employed the dorsal-air-pouch mouse model of inflammation and recorded an eight-fold increase in the recruitment of immune cells in mice treated with the probiotic strain, compared to the control group. Analysis of the exudates revealed significant changes in the expression of pro-inflammatory mediators on site. Treatment of Caco-2 cells with L. paracasei K5 induced significant upregulation of cytokines interleukin-1α (IL-1α), ΙL-1ß, IL-6, tumor necrosis factor-alpha (TNF-α), the chemokine C-X-C motif ligand 2 (CXCL2), and the inflammation markers soluble intercellular adhesion molecule (sICAM) and metallopeptidase inhibitor-1 (TIMP-1). Transient induction of the Toll-like receptors (TLRs) 2, 4, 6, and 9 expression levels was recorded by real-time PCR analysis. These results highlight the immunomodulatory potential of this strain and further support its probiotic character.

Functional pomegranate beverage production by fermentation with a novel synbiotic L. paracasei biocatalyst.

The recently isolated Lactobacillus paracasei K5 (a potential probiotic) was immobilized on delignified wheat bran (DWB) and was used to produce a functional pomegranate beverage. Fermentations were carried out for 24 h at different pH values (3.3, 3.6, and 3.9), and the fermented juices were stored for 4 weeks at 4 °C. In all cases, the immobilized biocatalyst was shown to be efficient for pomegranate juice fermentation, as indicated by analysis of sugars and organic acids, and ethanol production was maintained at low levels (0.5-1% v/v). The sugars consumed during fermentation were used mainly for cell mass production, while DWB exerted a prebiotic effect, protecting and enhancing the viability of L. paracasei. Optimum viability (9.8-11.7 log cfu/mL) was achieved at pH 3.9. The phenolic content and headspace volatiles were increased after fermentation and storage (112 ±â€¯17-213 ±â€¯30 mg GAE/100 mL and 27-46 compounds, respectively), indicating potential to produce good quality, synbiotic pomegranate beverages.

Combined Action of Piperitenone Epoxide and Antibiotics Against Clinical Isolates of Staphylococcus aureus and Escherichia coli.

The aim of this study was to determine the antimicrobial efficiency of piperitenone epoxide (PEO) - a principal component of various aromatic plants' essential oil - in combination with various antibiotics against 28 strains of Staphylococcus aureus and 10 strains of Escherichia coli isolated from clinical samples. Mentha spicata's essential oil, initially collected by hydrodistillation, was then subjected to flush column chromatography affording PEO of high purity. Minimum inhibitory concentrations of PEO alone and in combination with various concentrations of antibiotics were assessed using the microdilution method. The combined action was estimated calculating the fractional inhibitory concentration (FIC) index from checkerboard assays. Our results showed that the average minimum inhibitory concentration (mg/l) of PEO alone against E. coli was 512 ± 364.7 µg/ml, which was significantly higher than 172.8 ± 180.7 µg/ml observed for S. aureus. From checkerboard assays, FIC values below the 0.5 index, indicating synergy, were observed for 59% of the drugs tested. Twelve percent of FIC index values were between 0.5 and 1, indicating additive effects, while 21% were indifferent. According to our results, PEO could be a promising antimicrobial compound when combined with specific antibiotics and deserves further study.