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.

Chemical Composition, Antibacterial and Antibiofilm Actions of Oregano (Origanum vulgare subsp. hirtum) Essential Oil against Salmonella Typhimurium and Listeria monocytogenes.

Essential oils (EOs) are plant mixtures that are known to present strong bioactivities, including a wide antimicrobial action. Biofilms are microbial sessile structures that represent the default mode of growth of microorganisms in most environments. This study focused on the antimicrobial action of the EO extracted from one of the most representative oregano species, that is, Origanum vulgare (subsp. hirtum), against two important foodborne pathogens, Salmonella enterica (serovar Typhimurium) and Listeria monocytogenes. For this, the minimum inhibitory concentrations of the EO against the planktonic and biofilm growth of each bacterium were determined (MICs, MBICs), together with the minimum bactericidal and biofilm eradication concentrations (MBCs, MBECs). The EO was also analyzed for its chemical composition by gas chromatography-mass spectrometry analysis (GC-MS). The influence of EO exposure on the expression of some important virulence genes (hly, inlA, inlB and prfA) was also studied in L. monocytogenes. Results revealed a strong antibacterial and antibiofilm action with MICs and MBICs ranging from 0.03% to 0.06% (v/v) and from 0.06% to 0.13% (v/v), respectively. The application of the EO at 6.25% (v/v) for 15 min resulted in the total eradication of the biofilm cells of both pathogens. The EO was mainly composed of thymol, p-cymene, γ-terpinene and carvacrol. The 3 h exposure of L. monocytogenes planktonic cells to the EO at its MBIC (0.06% v/v) resulted in the significant downregulation of all the studied genes (p < 0.05). To sum, the results obtained advocate for the further exploitation of the antimicrobial action of oregano EO in food and health applications.

Novel Functional Grape Juices Fortified with Free or Immobilized Lacticaseibacillus rhamnosus OLXAL-1.

During the last decade, a rising interest in novel functional products containing probiotic microorganisms has been witnessed. As food processing and storage usually lead to a reduction of cell viability, freeze-dried cultures and immobilization are usually recommended in order to maintain adequate loads and deliver health benefits. In this study, freeze-dried (free and immobilized on apple pieces) Lacticaseibacillus rhamnosus OLXAL-1 cells were used to fortify grape juice. Juice storage at ambient temperature resulted in significantly higher (>7 log cfu/g) levels of immobilized L. rhamnosus cells compared to free cells after 4 days. On the other hand, refrigerated storage resulted in cell loads > 7 log cfu/g for both free and immobilized cells for up to 10 days, achieving populations > 109 cfu per share, with no spoilage noticed. The possible resistance of the novel fortified juice products to microbial spoilage (after deliberate spiking with Saccharomyces cerevisiae or Aspergillus niger) was also investigated. Significant growth limitation of both food-spoilage microorganisms was observed (both at 20 and 4 °C) when immobilized cells were contained compared to the unfortified juice. Keynote volatile compounds derived from the juice and the immobilization carrier were detected in all products by HS-SPME GC/MS analysis. PCA revealed that both the nature of the freeze-dried cells (free or immobilized), as well as storage temperature affected significantly the content of minor volatiles detected and resulted in significant differences in the total volatile concentration. Juices with freeze-dried immobilized cells were distinguished by the tasters and perceived as highly novel. Notably, all fortified juice products were accepted during the preliminary sensory evaluation.

Chemical Composition, Antioxidant, and Antibiofilm Properties of Essential Oil from Thymus capitatus Plants Organically Cultured on the Greek Island of Lemnos.

Essential oils (EOs) are mixtures of volatile plant secondary metabolites and have been exploited by humans for thousands of years for various purposes because of their many bioactivities. In this study, the EO from Thymus capitatus, a thyme species organically cultured on the Greek Island of Lemnos, was analyzed for its chemical composition (through GC-FID and GC-MS), antioxidant activity (AA), and total phenolic content (TPC), as well as its antimicrobial and antibiofilm actions against three important foodborne bacterial pathogens (Salmonella enterica ser. Typhimurium, Listeria monocytogenes, and Yersinia enterocolitica). For the latter investigations, the minimum inhibitory concentrations (MICs) and minimum biofilm inhibitory concentrations (MBICs) of the EO against the planktonic and biofilm growth of each pathogen were determined, together with the minimum biofilm eradication concentrations (MBECs). Results revealed that T. capitatus EO was rich in thymol, p-cymene, and carvacrol, presenting high AA and TPC (144.66 µmol TroloxTM equivalents and 231.32 mg gallic acid equivalents per g of EO, respectively), while its MICs and MBICs ranged from 0.03% to 0.06% v/v and 0.03% to 0.13% v/v, respectively, depending on the target pathogen. The EO was able to fully destroy preformed (mature) biofilms of all three pathogenic species upon application for 15 min, with MBECs ranging from 2.00 to 6.25% v/v. Overall, the results demonstrate that the EO of organically cultured T. capitatus presents strong antioxidant, antibacterial, and antibiofilm properties and could, therefore, be further exploited as a functional and antimicrobial natural formulation for food and health applications.

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.

Assessment of the Anti-Amyloidogenic Properties of Essential Oils and Their Constituents in Cells Using a Whole-Cell Recombinant Biosensor.

Essential oils exhibit numerous medicinal properties, including antimicrobial, anti-inflammatory and antioxidant effects. Recent studies also indicate that certain essential oils demonstrate anti-amyloidogenic activity against ß-amyloid, the protein implicated in Alzheimer's disease. To investigate whether the anti-aggregating properties of essential oils extend to α-synuclein, the protein involved in Parkinson's disease, we constructed and employed a whole-cell biosensor based on the split-luciferase complementation assay. We validated our biosensor by using baicalein, a known inhibitor of α-synuclein aggregation, and subsequently we tested eight essential oils commonly used in food and the hygienic industry. Two of them, citron and sage, along with their primary components, pure linalool (the main constituent in citron essential oil) and pure eucalyptol (1,8-cineole, the main constituent in sage essential oil), were able to reduce α-syn aggregation. These findings suggest that both essential oils and their main constituents could be regarded as potential components in functional foods or incorporated into complementary Parkinson's disease therapies.

Effect of Free or Immobilized Lactiplantibacillus plantarum T571 on Feta-Type Cheese Microbiome.

BACKGROUND: Cheese microbiome plays a key role in determining the organoleptic and physico-chemical properties and may be also used as an authenticity tool for distinguishing probiotic cultures. Due to significant reduction of cell viability often witnessed during food production processes and storage, immobilization is proposed to ascertain high probiotic cell loads required to confer the potential health benefits. Hence, the aim of the present study was to investigate the effect of free or immobilized Lactiplantibacillus plantarum T571 on whey protein on feta cheese microbiome. METHODS: Next-Generation Sequencing technology was used to investigate cheese microbiome. Cheese samples containing free or immobilized Lactiplantibacillus plantarum T571 (a wild type strain isolated from Feta cheese brine) on whey protein, along with products containing commercial starter culture, were analyzed. RESULTS: The results showed a great diversity of bacteria and fungi genera among the samples. An increased presence of Lactobacillus OTUs in cheese with immobilized cells on whey protein was witnessed, highlighting the survival of the strain in the final product. The immobilized culture had also a significant impact on other genera, such as Lactococcus, Leuconostoc and Debaryomyces, which are associated with improved technological characteristics and health benefits. CONCLUSIONS: Enrichment of feta cheese with immobilized potential probiotics to secure cell viability consists of an industrial challenge and leads to distinct microbiome composition that may be used as a valuable food authenticity tool.

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.

Impact of Sugar Type Addition and Fermentation Temperature on Pomegranate Alcoholic Beverage Production and Characteristics.

The present study focuses on the production of pomegranate alcoholic beverage (PAB) from juice of the Wonderful variety. The effect of fermentation temperature (15 and 25 °C) and type of sugar added (adjustment to 20 °Brix) on the physicochemical characteristics, bioactive compounds, and volatile composition were studied. Sucrose, concentrated pomegranate juice, concentrated grape juice, and honey were used to increase the initial sugar content. The produced PABs contained ethanol in concentrations ranging from 7.9 to 10.0% v/v and glycerol from 4.8 to 6.1 g L-1. A decrease in total phenolics content, free radical-scavenging activity, and total monomeric anthocyanin content was observed following fermentation. Total flavonoids content appeared to increase after fermentation only in the cases of concentrated pomegranate and grape juice addition. In general, 22 volatile compounds were identified in PABs (13 esters, 2 fatty acids, and 7 alcohols). Major compounds detected were 3-methyl-1-butanol, 2-methyl-1-butanol, 2-phenylethanol, and ethyl acetate. These findings demonstrate the production prospect of PABs with increased ethanol content, while elaborating on the importance of fermentation temperature and the differences between the selected types of added sugars on end-product composition.

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.

Evaluation of Yeast Strains for Pomegranate Alcoholic Beverage Production: Effect on Physicochemical Characteristics, Antioxidant Activity, and Aroma Compounds.

In the present study, three commercial yeasts (for wine, beer, and cider) were evaluated for the production of pomegranate alcoholic beverage (PAB) from a juice of Wonderful variety. The physicochemical characteristics, antioxidant activity, and aromatic profiles of PABs were investigated before and after fermentation, while the effect of yeast strain and fermentation temperature (15 and 25 °C) was also evaluated. The PABs contained ethanol in the ranges of 5.6-7.0% v/v, in combination with glycerol (2.65-6.05 g L-1), and low volatile acidity. Total flavonoid content, total phenolic content, free radical-scavenging activity, and total monomeric anthocyanin content appeared to decrease after fermentation, possibly due to hydrolysis, oxidation, and other reactions. In general, PABs retained 81-91% of free radical-scavenging activity, 29-41% of phenolics, 24-55% of flavonoids, and 66-75% of anthocyanins. The use of different yeast affected mainly flavonoids and anthocyanins, and yeast strain M02 resulted in the highest values after fermentation. In PABs, 30 different volatile compounds were identified, specifically 15 esters, 4 organic acids, 8 alcohols, and 3 terpenes. The principal component analysis showed that the fermentation temperature affected significantly volatile composition, whereas, among the yeasts, WB06 is the one that seems to differentiate. The findings of this study show that the selection of the appropriate yeast and fermentation temperature is very crucial and affects the characteristics of the final product.

Citrus medica and Cinnamomum zeylanicum Essential Oils as Potential Biopreservatives against Spoilage in Low Alcohol Wine Products.

Low alcohol wine is a new entry in the global wine market, due to the increase in consumers' concern for health, economic and modern lifestyle issues. As low alcohol products are prone to spoilage, the adoption of natural-derived products with antimicrobial activity as biopreservatives seems to be an intriguing alternative. Thus, the aim of the present study was to investigate the possible antimicrobial properties of Citrus medica and Cinnamomum zeylanicum essential oils (EOs) and assess their commercial prospective in the wine industry. The main constituents identified by GC/MS analysis were limonene (38.46%) and linalool (35.44%) in C. medica EO, whereas trans-cinnamic-aldehyde (63.58%) was the dominant compound in C. zeylanicum EO. The minimum inhibitory (MIC), non-inhibitory (NIC) and minimum lethal concentration (MLC) values against common wine spoilage microbes were initially determined. Subsequently, their efficiency was further validated in low alcohol (~6% vol) wines, either separately or in combination at 0.010% (v/v), as well as in wines deliberately inoculated with Gluconobacter cerinus, Oenococcus oeni, Pediococcus pentosaceus, Dekkera bruxellensis, Candida zemplinina, Hanseniaspora uvarum, Pichia guilliermondii or Zygosaccharomyces bailii. EO addition led to considerable spoilage and microbial growth delay during storage at room or refrigerated temperature, suggesting their potential use as wine biopreservatives.

Freeze-dried immobilized kefir culture in cider-making.

BACKGROUND: The aim of the present study was to evaluate the fermentation efficiency of freeze-dried immobilized kefir culture on natural supports (apple pieces, delignified cellulosic material) in cider making at various temperatures (5-45 °C) in comparison with freeze-dried free cells. Freeze-dried cells were initially tested in apple juice fermentations at 30 °C, and then the freeze-dried cultures produced with no cryoprotectants were assessed in repeated batch fermentations. RESULTS: Repeated batch fermentations lasted for longer than 5 months. High malic acid conversion rates (up to 78.5%) and ethanol productivity values (up to 37.9 g L-1 day-1 ) were recorded for freeze-dried immobilized cells. Polymerase chain reaction - denaturing gradient gel electrophoresis (PCR-DGGE) analysis showed that freeze-drying had no effect on the microbial diversity of kefir culture. Higher alcohols were significantly reduced at low fermentation temperatures. Application of principal component analysis (PCA) revealed that both the fermentation temperature and the nature of the freeze-dried kefir culture affected significantly the minor volatiles determined by gas chromatography/mass spectrometry (GC/MS). Notably, all ciders produced were of high quality and were accepted by the tasting panel. CONCLUSIONS: Freeze-dried immobilized kefir culture on natural supports with no cryoprotectants was found to be suitable for simultaneous alcoholic and malolactic cider fermentation at various temperatures (5-45 °C). The high operational stability of the systems was confirmed and the results obtained are of great interest for the industrial sector as they could be exploited for cider, low-alcohol cider, or 'soft' cider production. © 2020 Society of Chemical Industry.

Freeze-Dried Immobilized Kefir Culture in Low Alcohol Winemaking.

Low alcohol wines represent a rising trend in the global market. Since for ethanol removal, certain physicochemical methods that negatively affect wine quality are applied, the aim of this present study was to evaluate the efficiency of freeze-dried, immobilized kefir culture on natural supports (apple pieces, grape skins and delignified cellulosic material) in low alcohol winemaking at various temperatures (5-30 °C). Initially, genetic analysis of kefir culture was performed by Next Generation Sequencing. There was an immobilization of kefir culture on grape skins-enhanced cell survival during freeze-drying in most cases, even when no cryoprotectant was used. Simultaneous alcoholic and malolactic fermentations were performed in repeated batch fermentations for >12 months, using freeze-dried free or immobilized cells produced with no cryoprotectant, suggesting the high operational stability of the systems. Values of great industrial interest for daily ethanol productivity and malic acid conversion [up to 39.5 g/(Ld) and 67.3%, respectively] were recorded. Principal Component Analysis (PCA) showed that freeze-drying rather than the fermentation temperature affected significantly minor volatiles. All low alcohol wines produced were accepted during the preliminary sensory evaluation.

Wine production using free and immobilized kefir culture on natural supports.

The aim of the present study was to perform simultaneous alcoholic and malolactic wine fermentations using free or immobilized kefir culture at a wide temperature range (5-45 °C). Repeated batch fermentations were carried out for a period up to 29 months, suggesting a high operational stability of the systems, while malic acid conversion and ethanol productivity up to 70.9% and 36.9 g/(Ld) were noted. Volatile acidity was at levels typically found in wines (<1.0 g acetic acid/L) in most cases, although increased values were recorded in wines produced at 5 °C, but no vinegar taint was detected. Fusel alcohols were at levels usually found in wines, but were reduced at 5 °C. Application of PCA to minor volatiles showed that the fermentation temperature rather than the nature of kefir culture had a significant effect. Noticeably, all products were accepted by the panel during the preliminary sensory evaluation.

Exploitation of olive oil mill wastewaters and molasses for ethanol production using immobilized cells of Saccharomyces cerevisiae.

An alcoholic fermentation process is described, involving molasses, the main by-product of the sugar industry, blended with crude olive oil mill wastewaters (OOMWs) and immobilized Saccharomyces cerevisiae cells on delignified cellulosic material (DCM). For comparison, fermentations with free cells were also carried out. Initially, the optimum blending mixture for molasses dilution was sought after, while at a second step repeated batch fermentations at a temperature range 5-30 °C were performed to monitor the operational stability of the system. A 1/1 ratio of OOMWs/tap water blending mixture and cell immobilization resulted in higher fermentation parameters. Ethanol concentration and daily productivity values recorded at temperatures ≥ 20 °C (up to 67.8 g L-1 and 67.6 g L-1 d-1, respectively) could be adopted by the industrial sector, although the decline in fermentation efficiency observed, probably due to the toxicity effects of OOMWs. Finally, the potential of OOMWs treatment for ethanol production is highlighted and assessed.

A new methodology for rapid detection of Lactobacillus delbrueckii subsp. bulgaricus based on multiplex PCR.

In this study we present a novel multiplex PCR assay for rapid and efficient detection of Lactobacillus delbrueckii subsp. bulgaricus. The accuracy of our method was confirmed by the successful identification of L. delbrueckii subsp. bulgaricus in commercial yoghurts and food supplements and it may be readily applied to the food industry.