Minor bioactive lipids.

Bioactive lipids-major and minor-comprise an array of compounds belonging to different chemical categories. Among the minor bioactive lipids carotenoids, sterols and tocochromanols attract continuously the interest of food scientists, nutritionists and medical doctors for their importance in food processing, preservation and for their health properties. Provitamin A and non-provitamin A carotenoids are found in various food sources of plant and animal origin and are added to foods as colorants. Their interactions with other food ingredients are critical because of their role against reactive oxygen species. The role of cholesterol through the diet after decades of disputes is better justified whereas at the same time emphasis is given to the technological and health aspects of phytosterols, which became very efficiently part of the daily diet for many population groups. Last but not least the importance of vitamin E is in a continuous debate for over 100years whereas studies on tocotrienols are intensified as a result of a transient to palm oil product consumption globally. Chemistry, natural occurrence, absorption and metabolism, dietary intake and dietary recommendations, major health impacts and key technological issues are updated and discussed with the support of recent findings.

Effects of Dietary Supplementation of Pomegranate Peel with Xylanase on Egg Quality and Antioxidant Parameters in Laying Hens.

Pomegranate contains bioactive compounds in all its parts. In this study, two levels of pomegranate peel byproduct (PPB) with or without the inclusion of xylanase enzyme were used to supplement laying hens' diet, in a 2 × 2 full factorial design. A total of 48 Isa brown laying hens were fed the following experimental diets for 8 weeks: T1 (2.5% PPB); T2 (2.5% PPB and xylanase); T3 (5% PPB); T4 (5% PPB and xylanase). Eggs collected were analyzed for egg quality parameters. Moreover, egg yolks were analyzed for Malondialdehyde content (MDA), fatty acid profile and total phenolic content. The T2 eggs showed enhanced yolk coloration and greater yolk total phenolic content. The T3 and T4 egg yolks showed lower MDA levels compared with T1, T2. Overall, results have shown that (a) xylanase inclusion affected egg yolk coloration and total phenolic content when combined with 2.5% PPB dietary supplementation; (b) dietary supplementation of 5% PPB resulted in eggs with reduced MDA levels.

Xylanase Supplementation in Wheat-Based Diets of Laying Hens Affects the Egg Yolk Color, Carotenoid and Fatty Acid Profiles.

Wheat is rich in non-starch polysaccharides (NSP) and their degradation in poultry diets is promoted by exogenous carbohydrases. The objective here was to evaluate the effect of adding an intrinsically thermostable xylanase on wheat-based diets for laying hens in yolk color, carotenoid and fatty acid profiles of eggs. A total of 128 laying hens were used for 12 weeks. They were randomly allocated to four dietary treatments with different levels of xylanase: T1: control (no xylanase), T2: 30,000 U/g, T3: 45,000 U/g and T4: 90,000 U/g, with 32 birds, 16 replicates per treatment (2 birds/replicate). At the end of the experimental period, egg yolk color index, redness (a*) and yellowness (b*) of egg yolks were found significantly higher in all the enzyme supplemented diet groups (T2, T3, T4) compared with the control (T1). Canthaxanthin levels were significantly higher in T3 than T1 (p < 0.05). Total n-3, n-6 and total polyunsaturated fatty acids (FAs) were significantly higher in T4 compared with the control (p < 0.01), while the reverse trend was evidenced for monounsaturated FAs. Additionally, total n-3 FAs were higher in the T2 than T1 (p < 0.005). Overall, the results showed that exogenous xylanase enzyme supplementation in wheat-based diets for laying hens contribute to maintaining egg yolk color. Overall, exogenous xylanase enzyme supplemented at all levels in wheat-based laying hens' diets improved egg yolk color compared to the control diet. The enzyme supplemented at the higher level (90,000 U/g) improved polyunsaturated and reduced monounsaturated egg yolk fatty acid content.

Bay Laurel (Laurus nobilis L.) Essential Oil as a Food Preservative Source: Chemistry, Quality Control, Activity Assessment, and Applications to Olive Industry Products.

Essential oils (EOs) find application as flavoring agents in the food industry and are also desirable ingredients as they possess preservative properties. The Mediterranean diet involves the use of a lot of herbs and spices and their products (infusions, EOs) as condiments and for the preservation of foods. Application of EOs has the advantage of homogeneous dispersion in comparison with dry leaf use in small pieces or powder. Among them, Laurus nobilis (bay laurel) L. EO is an interesting source of volatiles, such as 1,8-cineole and eugenol, which are known for their preservative properties. Its flavor suits cooked red meat, poultry, and fish, as well as vegetarian dishes, according to Mediterranean recipes. The review is focused on its chemistry, quality control aspects, and recent trends in methods of analysis and activity assessment with a focus on potential antioxidant activity and applications to olive industry products. Findings indicate that this EO is not extensively studied in comparison with those from other Mediterranean plants, such as oregano EO. More work is needed to establish authenticity and activity methods, whereas the interest for using it for the preparation of flavored olive oil or for the aromatization and preservation of table oils must be further encouraged.

Strategic Priorities of the Scientific Plan of the European Research Infrastructure METROFOOD-RI for Promoting Metrology in Food and Nutrition.

The pan-European distributed Research Infrastructure for Promoting Metrology in Food and Nutrition (METROFOOD-RI) has evolved in the frame of the European Strategy Forum on Research Infrastructures (ESFRI) to promote high-quality metrology services across the food chain. The METROFOOD-RI comprises physical facilities and electronic facilities. The former includes Reference Material plants and analytical laboratories (the 'Metro' side) and also experimental fields/farms, processing/storage plants and kitchen-labs (the 'Food' side). The RI is currently prepared to apply for receiving the European Research Infrastructure Consortium (ERIC) legal status and is organised to fulfil the requirements for operation at the national, European Union (EU) and international level. In this view, the METROFOOD-RI partners have recently reviewed the scientific plan and elaborated strategic priorities on key thematic areas of research in the food and nutrition domain to which they have expertise to contribute to meet global societal challenges and face unexpected emergencies. The present review summarises the methodology and main outcomes of the research study that helped to identify the key thematic areas from a metrological standpoint, to articulate critical and emerging issues and demands and to structure how the integrated facilities of the RI can operate in the first five years of operation as ERIC.

On the Role of the Carboxyl Group to the Protective Effect of o-dihydroxybenzoic Acids to Saccharomyces cerevisiae Cells upon Induced Oxidative Stress.

In the present work, the role of the carboxyl group of o-dihydroxybenzoic acids (pyrocatechuic, 2,3-diOH-BA and protocatechuic, 3,4-diOH-BA) on the protection against induced oxidative stress in Saccharomyces cerevisiae was examined. Catechol (3,4-diOH-B) was included for comparison. Cell survival, antioxidant enzyme activities, and TBARS level were used to evaluate the efficiency upon the stress induced by H2O2 or cumene hydroperoxide. Theoretical calculation of atomic charge values, dipole moment, and a set of indices relevant to the redox properties of the compounds was also carried out in the liquid phase (water). Irrespective of the oxidant used, 2,3-diOH-BA required by far the lowest concentration (3-5 µM) to facilitate cell survival. The two acids did not activate catalase but reduced superoxide dismutase activity (3,4-diOH-BA>2,3-diOH-BA). TBARS assay showed an antioxidant effect only when H2O2 was used; equal activity for the two acids and inferior to that of 3,4-diOH B. Overall, theoretical and experimental findings suggest that the 2,3-diOH-BA high activity should be governed by metal chelation. In the case of 3,4-diOH BA, radical scavenging increases, and chelation capacity decreases. The lack of carboxyl moiety (3,4-diOH B) improves to radical scavenging, interaction with lipophilic free radicals, and antioxidant enzymes. The present study adds to our knowledge of the antioxidant mechanism of dietary phenols in biological systems.

On the Effect of Microwave Heating on Quality Characteristics and Functional Properties of Persimmon Juice and Its Residue.

In this study, it was investigated whether integration of microwave-heating into the pretreatment step of persimmon juice processing allows the concomitant production of both functional juice and added-value solid residue from the Diospyros Kaki "Jiro" cultivar. In this direction, persimmon pulp was treated under three different microwave-heating conditions (0.7, 4.2, and 8.4 kJ/g) prior to enzymatic maceration and compared to the non-heated material. Irrespective of microwave energy employed, the proposed hybrid treatment was highly efficient in terms of juice yield (70% w/w). The mildest heating conditions resulted in juice and residue that were both of inferior quality. Intensification of the microwave energy reduced the microbial load of the juice up to 2-log without compromising the content in total soluble solids, sugars, and L-ascorbic acid. Under the most drastic conditions, the juice was enriched in gallic acid, polyphenols, and potent DPPH● scavengers, but its orange color faded and was more acidic. In parallel, the solid juice residue retained pro-vitamin A carotenoids (~278 µg retinol activity equivalents) and low-methoxy pectin (9 g/100 g DW). Overall, our findings can assist the efforts of the local juice processing industry to utilize persimmon fruits through energy-efficient technologies in a sustainable approach.

Mechanism of antimicrobial activity of honeybee (Apis mellifera) venom on Gram-negative bacteria: Escherichia coli and Pseudomonas spp.

Honeybee venom (Apitoxin, BV), a secretion substance expelled from the venom gland of bees, has being reported as antimicrobial against various bacterial species; however, the mechanism of action remains uncharacterized. In this study, the antibacterial activity of BV was investigated on hygiene indicator Escherichia coli and the environmental pathogen and spoilage bacterial species, Pseudomonas putida and Pseudomonas fluorescens. An array of methods was combined to elucidate the mode of action of BV. Viability by culture on media was combined with assessing cell injury with flow cytometry analysis. ATP depletion was monitored as an indicator to metabolic activity of cells, by varying BV concentration (75, 225and 500 µg/mL), temperature (25 [Formula: see text] and 37 [Formula: see text]), and time of exposure (0 to 24 h). Venom presented moderate inhibitory effect on E. coli by viability assay, caused high membrane permeability and significant ATP loss where the effect was increased by increased concentration. The viability of P. putida was reduced to a greater extent than other tested bacteria at comparable venom concentrations and was dictated by exposure time. On the contrary, P. fluorescens appeared less affected by venom based on viability; however, flow cytometry and ATP analysis highlighted concentration- and time-dependent effect of venom. According to Transmission Electron Microscopy results, the deformation of the cell wall was evident for all species. This implies a common mechanism of action of the BV which is as follows: the cell wall destruction, change of membrane permeability, leakage of cell contents, inactivation of metabolic activity and finally cell death.

Flavored Olive Oil as a Preservation Means of Reduced Salt Spanish Style Green Table Olives (cv. Chalkidiki).

Reformulation of products fermented in brine is a challenging area of research. Continuing the efforts toward the establishment of table olives as a healthy food for all population groups, this study aimed at examining whether olive oil flavored with essential oils can be used as a preservation means for reduced salt Spanish style green table olives (cv. Chalkidiki). Response surface methodology was applied to organize experimentation and assess data. As independent factors, concentrations of the essential oils used (oregano, lemon balm and bay laurel) and time of storage under vacuum were set. Microbiological parameters (pathogens and fermentation-related microbes), color and firmness attributes were used as responses. Models indicated that each essential oil exerted a preservative role to maintain microbiological quality of reduced salt table olives. Concurrently, appearance attributes of the latter were retained at desirable values. Oregano essential oil had a profound role against pathogens. Lemon balm and bay laurel essential oils were found to be important for yeast population control. The results are promising toward the use of flavored olive oil as a preservation means for tailor-made reduced salt table olives, a practice that may enhance local industry innovative activity in a practical and effective way.

Natural ß-Carotene Production by Blakeslea trispora Cultivated in Spanish-Style Green Olive Processing Wastewaters.

In the current research, the potential of Spanish-style green olive processing wastewaters (lye and washing waters) exploitation toward natural ß-carotene production by Blakeslea trispora was tested for the first time. Mating culture generated by the joint cultivation of the heterothallic fungal strains ATCC 14271 and 14272 in the non-sterile lye and washing waters was able to grow, achieving the phytotoxic hydroxytyrosol degradation by 57.3% and 66.8%, respectively. However, the low sugar and nitrogen content of the streams did not favor carotenogenesis. Alternatively, in the nutrient-enriched effluents, a notable quantity of ß-carotene was produced, accounted for 61.2 mg/L (lye) and 64.1 mg/L (washing waters) (82-88% of total carotenoid content). Above all, enriched streams had a noteworthy stimulating effect on the ß-carotene synthesis, because both the maximum ß-carotene yield per volume of enriched effluents and specific ß-carotene production rate were higher when compared with the respective values obtained from trials with synthetic reference medium without added effluents. Hydroxytyrosol and tyrosol showed high stability during the non-sterile process for ß-carotene production by B. trispora grown in the enriched effluents. This finding strengthens the potential toward the generation of multiple high-value products, which could lower the natural ß-carotene production costs.

Processing Wastewaters from Spanish-Style cv. Chalkidiki Green Olives: A Potential Source of Enterococcus Casseliflavus and Hydroxytyrosol.

The purpose of this study was to examine the isolation of indigenous lactic acid bacteria (LAB) with functional properties from Spanish-style cv. Chalkidiki green olive processing wastewaters (GOW). Predominant indigenous LAB could serve as bioaugmentation agents/starter culture for table olives production and protected designation of origin specification. Spontaneous fermentation of fresh GOW over different temperatures (15 °C to 50 °C) and pH values (3.5 to 11.5) for 30 d enabled the isolation/molecular identification of the lactic acid bacterium Enterococcus casseliflavus and the plant-associated bacterium Bacillus amyloliquefaciens subsp. plantarum. E. casseliflavus was found to reduce chemical oxygen demand by 72%. Its resistance to extreme pH values, salinity, and temperature was successfully modeled and the minimum inhibitory concentration of oleuropein against the bacterial growth was determined (0.9 g/L). Furthermore, hydroxytyrosol content was doubled (up to 553 mg/L) after GOW spontaneous fermentation under acidic conditions at 15 °C to 30 °C for 120 d, creating an additional source of input. These results highlight the significance and potential of E. casseliflavus in Spanish-style cv. Chalkidiki green olive processing.

Feasibility of multi-hydrolytic enzymes production from optimized grape pomace residues and wheat bran mixture using Aspergillus niger in an integrated citric acid-enzymes production process.

In this study, the potential of the co-generation of hydrolytic enzymes in the biorefinery plant for citric acid fermentation was investigated. Aspergillus niger B60 mycelium along with the solid residue after the recovery of sugars from white pomace (WP') were recycled from citric acid fermentation. A mixture design was used to determine the optimum ternary feedstock mixture composed of WP' (15%), red grape pomace (15%) and wheat bran (70%) that produced the target enzymes with high activities, which were compared to those from pure feedstocks. Maximum carboxymethyl cellulase, polygalacturonase, amylase, xylanase and acid protease activities obtained through solid-state fermentation (120 h, 30 °C) of the feedstock mixture were 668 IU/g, 3,151 IU/g, 1,099 IU/g, 579 IU/g and 204 IU/g (dry mass basis), respectively. The system was successfully simulated in SuperPro Designer. Results showed that the enzymes production process serves as the main contributor to the profitability of the biorefinery plant.

Citric acid production from the integration of Spanish-style green olive processing wastewaters with white grape pomace by Aspergillus niger.

This study aimed to optimize an integrated simple process for citric acid production using Spanish-style green olive processing wastewaters enriched with sugars from white grape pomace and the robust Aspergillus niger B60. Mild mixing of equal quantities of the above streams governed satisfactory amount of appropriate carbon sources (equimolar mixture of glucose and fructose, 111.5 g/L) in the sugar-enriched wastewater and its neutralization. Various nutrients and fermentation conditions were investigated and maximum citric acid content (85 g/L) and yield (0.56 g/g) were obtained in liquid surface culture after minimum regulation by adding sucrose and NH4NO3 (100 g/L and 1.1 g/L, respectively). Scale-up experiments (5 L-scale) verified findings from small scale (250 mL). The chemical oxygen demand value and phenolic content of the treated wastewater were reduced by 78% and 64%, respectively. Findings support the potential for clustering the respective enterprises in a biorefinery plant for citric acid fermentation.

Changes in Phenolic Compounds and Phytotoxicity of the Spanish-Style Green Olive Processing Wastewaters by Aspergillus niger B60.

This study systematically investigated the degradation kinetics and changes in the composition of phenolic compounds in Spanish-style Chalkidiki green olive processing wastewaters (TOPWs) during treatment using Aspergillus niger B60. The fungal growth and phenol degradation kinetics were described sufficiently by the Logistic and Edward models, respectively. The maximum specific growth rate (2.626 1/d) and the maximum degradation rate (0.690 1/h) were observed at 1500 mg/L of total polar phenols, indicating the applicability of the process in TOPWs with a high concentration of phenolic compounds. Hydroxytyrosol and the other simple phenols were depleted after 3-8 days. The newly formed secoiridoid derivatives identified by HPLC-DAD-FLD and LC-MS are likely produced by oleoside and oleuropein aglycon via the action of fungal ß-glucosidase and esterase. The treated streams were found to be less phytotoxic with reduced chemical oxygen demand by up to 76%. Findings will provide useful information for the subsequent treatment of residual contaminants.

Balsamic type varietal vinegar from cv. Xinomavro (Northen Greece). Optimization and scale-up of the alcoholic fermentation step using indigenous multistarters.

Taguchi design was used to examine the effect of parameters that should be optimized in order to control the alcoholic fermentation of the concentrated grape must (CGM) from cv. Xinomavro using the best-performing indigenous Hanseniaspora uvarum and Saccharomyces cerevisiae strains as multistarters. The "optimum" combination of conditions (cell ratio of H. uvarum/S. cerevisiae; inoculum size and inoculation time of S. cerevisiae; fermentation time and temperature) resulted in an alcoholic product that meets ethanol (79 g/kg) and residual sugar (164 g/kg) content requirements for further use in the production of balsamic type vinegar. Multistarter fermentation affected positively the varietal organoleptic traits of the fermented CGM. 5-(Hydroxymethyl)-furfural content emerged as a critical factor for the standardization of this process. Scaling up experiments in 12 L barrels verified findings from small scale in 100 mL flasks. The results of this work can be used as a prototype in further similar efforts.

Functional Teas from the Leaves of Arbutus unedo: Phenolic Content, Antioxidant Activity, and Detection of Efficient Radical Scavengers.

The phenolic content/composition and antioxidant activity of hot/cold infusion and decoction from the leaves of Arbutus unedo were studied for the first time. 1,1-diphenyl-2-picrylhydrazyl (DPPH●), 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) radical cation (ABTS●+), crocin-bleaching, copper-reducing, and liposome accelerated oxidation assays were used for the evaluation of the activity in vitro. In vivo, the extracts were examined for their ability to protect S. cerevisiae cells from H2O2 induced oxidative stress. An on-line high-performance liquid chromatography-DPPH● assay was applied to identify potent radical scavengers and comment on their contribution to the total activity. The addition of leaves to boiling water (decoction) was the most appropriate practice to apply since the highest phenol intake (220.2 mg gallic acid/cup served) was obtained. Additionally, its antioxidant activity was equal or superior to that of the other extracts. Flavonols (~51-61 mg/g dry extract) were the main phenols in all the extracts, with quercitrin accounting for ~20% of the total phenol amount. The on-line DPPH● method verified the high potency of the decoction and indicated as the most active radical scavengers, two galloylquinic acid derivatives and myricitrin, accounting for ~28-45% and ~11-13% of the total scavenging, respectively. Present data may contribute to the future exploitation of A. unedo leaves by the food industry for health-promoting herbal tea preparations and dietary supplements.

Scale translation from shaken to diffused bubble aerated systems for lycopene production by Blakeslea trispora under stimulated conditions.

This study deals with the scale up of Blakeslea trispora culture from the successful surface-aerated shake flasks to dispersed-bubble aerated column reactor for lycopene production in the presence of lycopene cyclase inhibitor 2-methyl imidazole. Controlling the initial volumetric oxygen mass transfer coefficient (kLa) via airflow rate contributes to increasing cell mass and lycopene accumulation. Inhibitor effectiveness seems to decrease in conditions of high cell mass. Optimization of crude soybean oil (CSO), airflow rate, and 2-methyl imidazole was arranged according to central composite statistical design. The optimized levels of factors were 110.5 g/L, 2.3 vvm, and 29.5 mg/L, respectively. At this optimum setting, maximum lycopene yield (256 mg/L) was comparable or even higher to those reported in shake flasks and stirred tank reactor. 2-Methyl imidazole use at levels significantly lower than those reported for other inhibitors in the literature was successful in terms of process selectivity. CSO provides economic benefits to the process through its ability to stimulate lycopene synthesis, as an inexpensive carbon source and oxygen vector at the same time.

Implementing principles of traditional concentrated grape must fermentation to the production of new generation balsamic vinegars. Starter selection and effectiveness.

In an effort to implement principles of traditional concentrated grape must fermentation to the production of new generation balsamic vinegars (BVs), the specific goals of the study were the isolation and molecular identification of the predominant yeasts in concentrated grape must (cv. Xinomavro), their technological characterization and the evaluation of the fermentative aptitude of the selected strains. Tolerance against 5-hydroxymethyl-furfural (HMF) and furfural, acetic acid and glucose concentration was examined by appropriate methods and tests. The enological characteristics studied were acetic acid and H2S production, foaming and flocculation ability and key enzymatic activity. PCR-RFLP analysis revealed only the presence of Saccharomyces cerevisiae and Hanseniaspora uvarum among the 14 predominant osmophilic yeast isolates. Tolerance to both HMF and furfural was found strain- and dose-dependent and was suggested as a critical factor in the pre-selection of yeast starters. The most tolerant yeasts to these stress factors, a S. cerevisiae and a non-Saccharomyces strains, showed satisfactory growth in the presence of high glucose and acetic acid content (up to 600 g/L and 2 % w/w, respectively) and desirable enological characteristics. Results from the comparative evaluation of the fermentative aptitude of these strains with a commercial wine strain highlighted that the isolates had glucophilic behaviour and ability to produce desirable amounts of ethanol (100-120 g/kg) in short time (~20 d). The key volatiles useful for varietal discrimination and differentiation between the BVs and the traditional ones were also evaluated.

Bioflavour production from orange peel hydrolysate using immobilized Saccharomyces cerevisiae.

The rising trend of bioflavour synthesis by microorganisms is hindered by the high manufacturing costs, partially attributed to the cost of the starting material. To overcome this limitation, in the present study, dilute-acid hydrolysate of orange peel was employed as a low-cost, rich in fermentable sugars substrate for the production of flavour-active compounds by Saccharomyces cerevisiae. With this purpose, the use of immobilized cell technology to protect cells against the various inhibitory compounds present in the hydrolysate was evaluated with regard to yeast viability, carbon and nitrogen consumption and cell ability to produce flavour active compounds. For cell immobilization the encapsulation in Ca alginate beads was used. The results were compared with those obtained using free-cell system. Based on the data obtained immobilized cells showed better growth performance and increased ability for de novo synthesis of volatile esters of "fruity" aroma (phenylethyl acetate, ethyl hexanoate, octanoate, decanoate and dodecanoate) than those of free cells. The potential for in situ production of new formulations containing flavour-active compounds derive from yeast cells and also from essential oil of orange peel (limonene, α-terpineol) was demonstrated by the fact that bioflavour mixture was found to accumulate within the beads. Furthermore, the ability of the immobilized yeast to perform efficiently repeated batch fermentations of orange peel hydrolysate for bioflavour production was successfully maintained after six consecutive cycles of a total period of 240 h.

Revisiting extraction of bioactive apocarotenoids from Crocus sativus L. dry stigmas (saffron).

An ultrasound assisted extraction method is proposed for the recovery of bioactive glycosides (i.e. crocins and picrocrocin) from Crocus sativus L. dry stigmas using aqueous methanol. Response surface methodology (RSM) was employed to optimize the extraction parameters, namely, the percentage of methanol (%), the duration (min) and the duty cycles (s) of sonication. Optical microscopy, spectrophotometry and RP-HPLC-DAD were employed to follow pros and cons of the process. Additional experiments were conducted to compare recoveries with those under other agitation conditions (e.g. magnetic stirring according to ISO 3632-2 standard). The percentage of methanol, the sonication duration and duty cycles combination that can be recommended as optimum for the recovery of crocins and picrocrocin were 50%, 30 min, 0.2s and 0.44%, 30 min, 0.6s, respectively. Picrocrocin levels were not influenced dramatically under the optimum conditions for crocins extraction (11±2 instead of 12±1 mg kg(-1) dry stigmas, respectively) so that these can be considered optimum for both categories of tested compounds. Ultrasound assisted extraction speeded up further recovery of these precious apocarotenoids. Our findings for extraction conditions are useful for both industrial and analytical applications and should be considered in a forthcoming revision of the ISO 3632-2 technical standard.