Nutritional upgrade of olive mill stone waste, walnut shell and their mixtures by applying solid state fermentation initiated by Pleurotus ostreatus.

Present study concerns the transformation of the agro-industrial by-products olive mill stone waste (OMSW) and walnut shell (WS) to a protein-enriched animal feedstuff utilizing the solid state fermentation (SSF) technique. For this purpose, various mixtures of these by-products were exploited as substrates of the SSF process which was initiated by the P. ostreatus fungus. The respective results indicated that the substrate consisted of 80% WS and 20% OMSW afforded the product with the highest increase in protein content, which accounted the 7.57% of its mass (69.35% increase). In addition, a 26.13% reduction of lignin content was observed, while the most profound effect was observed for their 1,3-1,6 ß-glucans profile, which was increased by 3-folds reaching the 6.94% of substrate's mass. These results are indicative of the OMSW and WS mixtures potential to act as efficient substrate for the development of novel proteinaceous animal feed supplements using the SSF procedure. Study herein contributes to the reintegration of the agro-industrial by-products aiming to confront the problem of proteinaceous animal feed scarcity and reduce in parallel the environmental footprint of the agro-industrial processes within the context of circular economy.

Improved Antioxidant Blood Parameters in Piglets Fed Diets Containing Solid-State Fermented Mixture of Olive Mill Stone Waste and Lathyrus clymenum Husks.

Solid-state fermentation represents a sustainable approach for the conversion of agro-industrial wastes into high-added-value feed ingredients. The present study aimed to evaluate the effects of the dietary addition of a solid-state-fermented mixture of olive mill stone waste (OMSW) and Lathyrus clymenum husks (LP) on the antioxidant blood parameters of weaned piglets. Two hundred 35-day-old weaned piglets were allotted into two groups and fed either a control (C) diet or a diet containing 50 g of OMSW-LP per kg (OMSW-LP) for 40 days. Blood samples were collected at 35 and 75 days of age to assess the free radical scavenging activity (FRSA), reduced glutathione (GSH) levels, catalase activity (CAT), protein carbonyls (CARBs), and thiobarbituric acid reactive species (TBARS). The OMSW-LP diet reduced the TBARS (p = 0.049) and CARB contents (p = 0.012) and increased the levels of FRSA (p = 0.005), GSH (p = 0.040), and CAT activity (p = 0.012) in the piglets' blood, likely due to the synergistic action of the antioxidants and bioactive compounds present in the OMSW-LP mixture. Overall, the dietary inclusion of solid-state-fermented OMSW-LP at 50 g/kg could potentially serve a bio-functional purpose since it enhanced the antioxidant blood parameters in this study, a crucial factor for the health and growth of piglets post-weaning.

Unlocking the potential of spent coffee grounds via a comprehensive biorefinery approach: production of microbial oil and carotenoids under fed-batch fermentation.

The valorization of renewable feedstock to produce a plethora of value-added products could promote the transition towards a circular bioeconomy. This study presents the development of cascade processes to bioconvert spent coffee grounds (SCGs) into microbial oil and carotenoids employing sustainable practices. The stepwise recovery of crude phenolic extract and coffee oil was carried out using green or recyclable solvents, i.e., aqueous ethanol and hexane. Palmitic acid (43.3%) and linoleic acid (38.9%) were the major fatty acids in the oil fraction of SCGs. The LC-MS analysis of crude phenolic extracts revealed that chlorogenic acid dominated (45.7%), while neochlorogenic acid was also detected in substantial amounts (24.0%). SCGs free of coffee oil and phenolic compounds were subjected to microwave-assisted pretreatment under different irradiations and solvents to enhance subsequent enzymatic saccharification. Microwave/water pretreatment at 400 W, followed by enzymatic hydrolysis with proteases, hemicellulases, and cellulases, at 50 g/L initial SCGs, led to satisfying overall yields of cellulose (75.4%), hemicellulose (50.3%), and holocellulose (55.3%). Mannan was the most extractable polysaccharide followed by galactan and arabinan. SCGs hydrolysate was used in fed-batch bioreactor fermentations with Rhodosporidium toruloides to produce 24.0 g/L microbial oil and carotenoids of 432.9 µg/g biomass.

Εleven Greek Legume Beans: Assessment of Genotypic Effect on Their Phytochemical Content and Antioxidant Properties.

Legumes, one of the first crops of humanity, inherently constitute a staple nutritional source for mankind, attracting significant research attention that has been afforded to the development of numerous cultivars. The study herein concerns the exploitation of the nutritional and bio-functional content of beans harvested from eleven Greek cultivars belonging to five different species, namely Cicer arietinum L., Pisum sativum L., Vicia faba L., Lens culinaris L., and Phaseolus vulgaris L. The final goal is to define their varietal identity and correlate their phytochemical content with their potential utilization as functional foods and/or feed of high nutritional value. In this respect, their extracts were screened against the presence of 27 fatty acids and 19 phenolic compounds, revealing the presence of 22 and 15 molecules, respectively. Specifically, numerous fatty acids were detected in significant amounts in all but C. arietinum extract, while significant polyphenolic content was confirmed only in P. vulgaris. Among individual compounds, linoleic acid was the major fatty acid detected in amounts averaging more than 150 mg/g, followed by oleic acid, which was present as a major compound in all extracts. Among the nine polyphenols detected in P. vulgaris, the molecules of genistein (3.88 mg/g) and coumestrol (0.82 mg/g) were the most abundant. Their antioxidant properties were evaluated through DPPH and FRAP assays, which were highlighted as most potent in both tests of the V. faba extract, while C. arietinum was determined as totally inactive, indicating a potential correlation between the phenolic content of the plant species and antioxidant activity. These results are indicative of the significant advances achieved for the cultivars investigated and reveal their important role as nutritional crops for human and animal consumption.

Assessment of the volatile and non-volatile profile of Savatiano PGI wines as affected by various terroirs in Attica, Greece.

Establishing the concept of terroir in wines, combined with the exploitation of native grape varieties, is considered a viable alternative to produce quality wines for increasing interest in the wine market. The aim of this study is the characterization of Protected Geographical Indication (PGI) Savatiano white wines from different regions of Attica (Greece), through the chemical and sensory description of the aroma of wines and the determination of their phenolic profiles. The wines produced with the same vinification protocol were evaluated using the descriptive sensory analysis method while they underwent profiling of volatile and phenolic compounds by gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) respectively. The presence of regional differences within the wines was also explored, elaborated with variables that contribute to their differentiation, such as soil type, monthly average temperature, and rainfall. Volatile and sensory profiles of the wines separated the regions and confirmed that sub-regional differences attributed mainly to soil characteristics affect wine aroma. The wines from the east part of Attica were higher in esters, terpenes and higher alcohols with high scores in fruity and blossoms attributes while wines from the north part of Attica presented higher intensity of mineral, nutty and herbaceous attributes. The separations based solely on the phenolic compounds concentration were less clear but a relationship was found between the content of phenolic acids and flavonoids and the studied regions. To our knowledge this is the first characterization of Savatiano PGI wines of Attica generating a fingerprint including chemical composition and sensory aroma characteristics to differentiate wines, combining this pattern with particular sub-regions.

An Interdisciplinary Assessment of Biochemical and Antioxidant Attributes of Six Greek Vicia sativa L. Varieties.

Common vetch (Vicia sativa L.) is one of the most cultivated feed crops with extensive agricultural diversity and numerous cultivars. This study concerns the first-time investigation of the dry plant biomass and grains of six vetch cultivars to define the detailed fingerprint of their phenolic and fatty acid content, along with their respective antioxidant potencies. The results revealed a substantial variation in the feed quality traits among the tested Vicia sativa varieties, highlighting the crucial role and influence the genotype plays in the achievement of high-quality livestock nutrition. Among the six varieties tested, Istros and M-6900 displayed a particularly intriguing phytochemical profile characterized by elevated phenolic content, significant antioxidant potency and remarkably high fatty acid indices. These findings are indicative of the great potential of these varieties to function as suitable candidates for incorporation into farm animal diets either in the form of dry biomass (hay) or as a grain feed additive.

Phytoestrogens as Biomarkers of Plant Raw Materials Used for Fish Feed Production.

The intensive use of plant materials as a sustainable alternative for fish feed production, combined with their phytochemical content, which affects the growth and production characteristics of farmed fishes, necessitates their monitoring for the presence of raw materials of plant origin. This study reported herein concerns the development, validation and application of a workflow using high-performance liquid chromatography combined with tandem mass spectrometry (LC-MS/MS) for the quantification of 67 natural phytoestrogens in plant-derived raw materials that were used to produce fish feeds. Specifically, we verified the presence of 8 phytoestrogens in rapeseed meal samples, 20 in soybean meal samples, 12 in sunflower meal samples and only 1 in wheat meal samples in quantities enabling their efficient incorporation into clusters. Among the various constituents, the soybean phytoestrogens daidzein, genistein, daidzin, glycitin, apigenin, calycosin and coumestrol, as well as the sunflower neochlorogenic, caffeic and chlorogenic phenolic acids, displayed the highest correlations with their origin descriptions. A hierarchical cluster analysis of the studied samples, based on their phytoestrogen contents, led to the efficient clustering of raw materials. The accuracy and efficiency of this clustering were tested through the incorporation of additional samples of soybean meal, wheat meal and maize meal, which verified the utilization of the phytoestrogen content as a valuable biomarker for the discrimination of raw materials used for fish feed production.

Farm or Lab? A Comparative Study of Oregano's Leaf and Callus Volatile Isolates Chemistry and Cytotoxicity.

Oregano (Origanum vulgare, Lamiaceae plant family) is a well-known aromatic herb with great commercial value, thoroughly utilized by food and pharmaceutical industries. The present work regards the comparative assessment of in vitro propagated and commercially available oregano tissue natural products. This study includes their secondary metabolites' biosynthesis, antioxidant properties, and anticancer activities. The optimization of callus induction from derived oregano leaf explants and excessive oxidative browning was performed using various plant growth regulators, light conditions, and antioxidant compounds. The determination of oregano callus volatiles against the respective molecules in maternal herbal material was performed using gas chromatography-mass spectrometry (GC/MS) analysis. In total, the presence of twenty-seven phytochemicals was revealed in both leaf and callus extracts, from which thirteen molecules were biosynthesized in both tissues studied, seven compounds were present only in callus extracts, and seven metabolites only in leaf extracts. Carvacrol and sabinene hydrate were the prevailing volatiles in all tissues exploited, along with alkanes octacosane and triacontane and the trimethylsilyl (TMS) derivative of carvacrol that were detected in significant amounts only in callus extracts. The MTT assay was employed to assess the in vitro cytotoxic properties of oregano extracts against the epithelial human breast cancer MDA-MB-231 and the human neuroblastoma SK-N-SH cell lines. The extracts displayed concentration and time-dependent responses in cell proliferation rates.

Bioactivity of Wild and Cultivated Legumes: Phytochemical Content and Antioxidant Properties.

The global demand for increased meat production has brought to the surface several obstacles concerning environmental impacts, animals' welfare, and quality features, revealing the need to produce safe foodstuffs with an environmentally acceptable procedure. In this regard, the incorporation of legumes into animal diets constitutes a sustainable way out that prevents these apprehensions. Legumes are plant crops belonging to the Fabaceae family and are known for their rich content of secondary metabolites., displaying significant antioxidant properties and a series of health and environmental benefits. The study herein aims to investigate the chemical composition and antioxidant activities of indigenous and cultivated legume plants used for food and feed. The respective results indicate that the methanolic extract of Lathyrus laxiflorus (Desf.) Kuntze displayed the highest phenolic (64.8 mg gallic acid equivalents/g extract) and tannin (419.6 mg catechin equivalents/g extract) content, while the dichloromethane extract of Astragalus glycyphyllos L., Trifolium physodes Steven ex M.Bieb. and Bituminaria bituminosa (L.) C.H.Stirt. plant samples exhibited the richest content in carotenoids lutein (0.0431 mg/g A. glycyphyllos extract and 0.0546 mg/g B. bituminosa extract), α-carotene (0.0431 mg/g T. physodes extract) and ß-carotene (0.090 mg/g T. physodes extract and 0.3705 mg/g B. bituminosa extract) establishing their potential role as vitamin A precursor sources. Results presented herein verify the great potential of Fabaceae family plants for utilization as pasture plants and/or dietary ingredients, since their cultivation has a positive impact on the environment, and they were found to contain essential nutrients capable to improve health, welfare, and safety.

Growth and Volatile Organic Compound Production of Pseudomonas Fish Spoiler Strains on Fish Juice Agar Model Substrate at Different Temperatures.

Microbial spoilage is the main cause of quality deterioration in seafood. Several strains of psychotropic Pseudomonas have been found to dominate in such products, producing a plethora of volatile organic compounds (VOC). Herein, we investigated the growth of and VOC production by seven strains of Pseudomonas associated with spoiled fish after inoculation as single and mixed cultures on model fish substrate and storage at 0, 4 and 8 °C. The results indicated a strain-dependent VOC profile that was also affected by the storage temperature. Hierarchical cluster analysis (HCA) successfully grouped the strains based on VOC profile at each studied temperature, while some potential Chemical Spoilage Indices (CSI) were revealed. The findings of the present work will contribute to the understanding of the metabolic activity of particular strains of Pseudomonas and to reveal any potential CSI for rapid evaluation of fish spoilage/freshness status.

Pigments' analysis of Citrus juicing making by-products by LC-MS/MS and LC-DAD.

Citrus fruits Cold Press Essential Oils (CPEOs) constitute a low-cost by-product produced during the juice making process that are composed by a volatile and a non-volatile fraction. Their non-volatile fractions are rich in valuable secondary metabolites, such as carotenoids, coumarins, psoralens and flavonoids [1,2]. Study herein concerns the development of a quantitative method for their carotenoids analyses using the LC-MS/MS and LC-DAD methodology.•CPEOs carotenoids content was characterized quantitatively.•Natural carotenoids ß-cryptoxanthin, lutein and zeaxanthin were determined as the most abundant high added value molecules.

Efficacy Evaluation of Oregano Essential Oil Mixed with Bacillus thuringiensis israelensis and Diflubenzuron against Culex pipiens and Aedes albopictus in Road Drains of Italy.

Mosquito management programs in the urban environment of Italian cities mainly rely on larval control with conventional insecticides, primarily targeting the road drains that constitute the principal mosquito breeding sites encountered in public. The repeated utilization of synthetic insecticides may have adverse effects on non-targets and lead to resistance development issues, while the performance of biopesticides encounters limitations in field use. Botanical insecticides as single larval control agents or in binary mixtures with conventional insecticides have been extensively studied in the laboratory as an effective and eco-friendly alternative mosquito control method with promising results. The study herein concerns the investigation, for the first time under realistic conditions in the field, of the joint action of a carvacrol-rich oregano Essential Oil (EO) with two conventional insecticides, namely, the insect growth regulator diflubenzuron and the bio-insecticide Bacillus thuringiensis israelensis (B.t.i.), in road drains in Crevalcore city, Italy, against Culex pipiens and Aedes albopictus. According to the obtained results, the application of both plain EO and its mixtures with diflubenzuron and B.t.i. exerted very high efficacy in terms of immature mosquito population reduction over a two-week period. Three weeks after treatment, the performance of the oil and its mixtures diminished but remained high, while the addition of diflubenzuron potentiated the persistent action of the oil against Cx. pipiens. These findings are indicative of the potential of mixing carvacrol-rich EO with diflubenzuron and B.t.i. as an efficient eco-friendly alternative to mono-insecticide applications in road drains against Cx. pipiens and Ae. albopictus larvae.

Transformation of mixtures of olive mill stone waste and oat bran or Lathyrus clymenum pericarps into high added value products using solid state fermentation.

The main objective of this study concerns the bioconversion of agro-industrial wastes into high added value products, such as proteinaceous animal feed, using Solid State Fermentation process (SSF). For this purpose, the Olive Mill Stone Waste (OMSW) which is known to contain low amounts of proteins and a high concentration of anti-nutritional substances was used as substrate. Subsequently, OMSW was fermented with Oat Bran (OB) or Lathyrus clymenum pericarp (LP) in proportions varying from 10 %w/w to 30 %w/w, applying SSF process initiated by Pleurotus ostreatus utilizing latter's secreted enzymes for their degradation. The respective results indicated that the addition of 30 %w/w of OB, resulted in a 39% increase of the protein content at the end of fermentation (Day 21). In addition, we observed a 5-fold increase of 1,3-1,6 ß-glucan content and a simultaneous decrease of unwanted lignin of 24%. The addition of 20 %w/w of LP afforded, at the end of fermentation (Day 21), an increased protein content of 57%, a 3-fold increase in 1,3-1,6 ß-glucans and a decrease in lignin concentration of 13%. These findings demonstrate the potential of the LP utilization by using SSF process, for the production of high nutritional value dietary supplements for animal feed. This endeavor constitutes the first literature report for the utilization of the agro-industrial waste LP. The developed methodology herein is considered as crucial for the circular economy since it refers to the reuse of agro-industrial wastes and the production of a high added-value product.

Essential Oil Coating: Mediterranean Culinary Plants as Grain Protectants against Larvae and Adults of Tribolium castaneum and Trogoderma granarium.

Postharvest agricultural losses constitute a major food security risk. In contrast, postharvest protection is strongly linked with food safety. The present study aims to develop novel postharvest protection tools through a bioprospecting protocol utilizing edible essential oils (EOs) as grain coatings. For this purpose, six Mediterranean culinary plants were selected for evaluation. The EOs of juniper, Juniperus phoenicea L. (Pinales: Cupressaceae), marjoram, Origanum majorana L. (Lamiales: Lamiaceae), oregano, Origanum vulgare ssp. hirtum (Link) A.Terracc. (Lamiales: Lamiaceae), bay laurel, Laurus nobilis L. (Laurales: Lauraceae) and tarhan, Echinophora tenuifolia ssp. sibthorpiana (Guss.) Tutin (Apiales: Apiaceae) were retrieved through steam distillation, while lemon, Citrus limon (L.) Osbeck (Sapindales: Rutaceae) EO was retrieved through cold press extraction. All EOs were formulated to microemulsions (MEs) and applied uniformly as a coating on wheat against larvae and adults of Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae) and Trogoderma granarium Everts (Coleoptera: Dermestidae). All EO-based MEs have been evaluated for the first time as grain coatings. They caused moderate to high mortality to T. castaneum larvae (67.8-93.3% 14 days post-exposure) and T. granarium adults (70.0-87.8% after 7 days of exposure). Citrus limon, O. majorana and E. tenuifolia ssp. sibthorpiana EO-based MEs were the most efficient against T. castaneum larvae, by exhibiting 93.3%, 91.1% and 90.0% mortality 14 days post-exposure, respectively. Origanum majorana, L. nobilis and J. phoenicea EO-based MEs were the most efficient against T. granarium adults, exhibiting 87.8%, 84.4% and 83.3% mortality after 7 days of exposure, respectively. These results indicate that EO-based ME coating is a potent tool against the tested postharvest pests.

Effect of a Carotenoid Extract from Citrus reticulata By-Products on the Immune-Oxidative Status of Broilers.

Although carotenoids generally possess antimicrobial and antioxidant properties, the in vivo synergistic action of carotenoid blends derived from plant-based by-products has not been thoroughly studied. Therefore, the carotenoid characterization and antimicrobial potential of Citrus reticulata extract as well as the impact of this carotenoid-rich extract (CCE) dietary supplementation on the performance, meat quality, and immune-oxidative status of broiler chickens were determined. One hundred and twenty one-day-old hatched chicks (Ross 308) were allocated to two dietary groups, with four replicate pens of 15 birds each. Birds were fed either a basal diet (CON) or the basal diet supplemented with 0.1% CCE (25 mg carotenoid extract included in 1 g of soluble starch) for 42 d. ß-Cryptoxanthin, ß-Carotene, Zeaxanthin, and Lutein were the prevailing carotenoid compounds in the Citrus reticulata extract. The CCE feed additive exerted inhibitory properties against both Gram-positive (Staphylococcus aureus) and negative (Klebsiella oxytoca, Escherichia coli, and Salmonella typhimurium) bacteria. Both the broiler performance and meat quality did not substantially differ, while the breast muscle malondialdehyde (MDA) concentration tended to decrease (p = 0.070) in the CCE-fed broilers. The inclusion of CCE decreased the alanine aminotransferase and MDA concentration, and the activity of glutathione peroxidase, while the activity of superoxide dismutase was increased in the blood. Catalase and NADPH oxidase 2 relative transcript levels were significantly downregulated in the livers of the CCE-fed broilers. Additionally, Interleukin 1ß and tumor necrosis factor (TNF) relative transcript levels were downregulated in the livers of the CCE- fed broilers, while TNF and interferon γ (IFNG) tended to decrease in the spleens and bursa of Fabricius, respectively. The present study provided new insights regarding the beneficial properties of carotenoids contained in Citrus reticulata in broilers' immune-oxidative status. These promising outcomes could be the basis for further research under field conditions.

Integrated biorefinery development using winery waste streams for the production of bacterial cellulose, succinic acid and value-added fractions.

An integrated biorefinery has been developed using winery wastes (grape pomace-GP, stalks-GS, wine lees-WL). Bacterial cellulose was produced from GP extracted free sugars. Grape-seed oil and polyphenols were extracted from GP. Experimental design was employed to optimize lignin removal (50.8%) from mixtures of remaining GP solids and GS via NaOH (1.19% w/v) treatment at 70°C for 30 min. Delignification liquid contained condensed tannins with 76% Stiasny number. Enzymatic hydrolysis produced a sugar-rich hydrolysate (40.2 g/L sugars). Ethanol, antioxidants, tartaric acid and nutrient-rich hydrolysate were produced from WL. The crude hydrolysates were used in fed-batch Actinobacillus succinogenes cultures for 37.2 g/L succinic acid production. The biorefinery produces 42.65 g bacterial cellulose, 24.3 g oil, 40.3 g phenolic-rich extract with 1.41 Antioxidant Activity Index, 80.2 g ethanol, 624.8 g crude tannin extract, 20.03 g tartaric acid and 157.8 g succinic acid from 1 kg of each waste stream.

Conversion of brewers' spent grain into proteinaceous animal feed using solid state fermentation.

Brewers' spent grain (BSG) represents the 85% of the total residue produced during the beer brewing process, with a global annual production volume exceeding 30 Mtons. The current study concerns the application of solid state fermentation (SSF) as a bioprocess where the nutritional value of BSG is improved for further use as animal feed with increased value. The investigated SSF procedure was initiated by the edible fungi Pleurotus ostreatus, which constitutes a natural source of proteins, ß-glucans, and various metabolites (vitamins, nutrients, etc.). Herein, the SSF of BSG resulted in a significant increase of protein content by 49.49%, a 10-fold increase of 1,3-1,6 ß-glucans, and a respective reduction of cellulose by 11.42%. The application of this method is expected to provide some useful information on the utilization of BSG as substrate for fungi-initiated SSF, a bioprocess allowing the significant reduction of the environmental impact caused by the beer brewing industry and simultaneously producing animal feed with higher protein content and improved nutritional characteristics. Such studies contribute to confront the unavailability of proteinaceous animal feed observed in the last decade.

Essential Oils Land Footprint: A Sustainability Meta-Analysis of Essential Oils Biopesticides.

BACKGROUND: Essential oils (EO) are considered as safe and sustainable alternatives of synthetically produced industrial raw materials. While EO are renewable resources their production is traced to land use, therefore employing nonrenewable resources. This fact is often neglected during market up-take, which is established on EO bioactivity efficacy. METHODS: Present study is aiming this knowledge gap through an innovative algorithm that employs spatial yield, bioactivity performance and fundamental experimentation details to calculate the land footprint. The proposed methodology is tested upon a concise pool of 54 EO, of which 9 originate from 8 culinary herbs, 27 from 3 juniper taxa, and 18 from 6 Citrus sp. crops. All 54 EO were subjected to repellent evaluation and 44 of them also to larvicidal, encompassing in the protocol both choice and no-choice bioassays. RESULTS: Based on these bioprospecting data the proposed protocol effectively calculated the land footprint for all EO and bioassays. The repellent land footprint indicated as more sustainable the EO from savory, oregano, tarhan, thyme, Greek sage, and juniper berries for which each application corresponds to 3.97, 4.74, 7.33, 7.66, 8.01 and 8.32 m2 respectively. The larvicidal assessment suggested as more sustainable the EOs from savory, oregano, fennel, thyme, tarhan, and rue with land footprints of 1.56, 1.79, 2.16, 2.89, 3.70 and 4.30 m2 respectively. CONCLUSIONS: The proposed protocol managed to calculate the land footprint for each EO and bioactivity and indicated the more sustainable EO per use based on widely available bioprospecting data.

Retrieval of High Added Value Natural Bioactive Coumarins from Mandarin Juice-Making Industrial Byproduct.

Cold pressed essential oil (CPEO) of mandarin (Citrus reticulata Blanco), a by-product of the juice-making industrial process known to contain large amounts of polymethoxyflavones, was exploited for its content in high added value natural coumarins. The study herein afforded a method referring to the evaporation of CPEO volatile fraction under mild conditions (reduced pressure and temperature below 35 °C) as azeotrope with isopropanol. This allowed the isolation of high added value coumarins from the non-volatile fragment using preparative High Performance Liquid Chromatography (HPLC). Pilot-scale application of this procedure afforded for each kg of CPEO processed the following natural bioactive coumarins in chemically pure forms: heraclenol (38-55 mg), 8-gerayloxypsoralen (35-51 mg), auraptene (22-33 mg), and bergamottin (14-19 mg). The structures of coumarins were verified by Nuclear Magnetic Resonance (NMR) spectroscopy and HPLC co-injection with authentic standards. Thus, the low market value mandarin CPEO with current value of 17 to 22 EUR/kg can be valorized through the production of four highly bioactive natural compounds worth 3479 to 5057 EUR/kg, indicating the great potentials of this methodology in the terms of the circular economy.

Evaluation of antimicrobial activities of plant aqueous extracts against Salmonella Typhimurium and their application to improve safety of pork meat.

Nine odorless laboratory-collected hydro-distilled aqueous extracts (basil, calendula, centrifuged oregano, corn silk, laurel, oregano, rosemary, spearmint, thyme) and one industrial steam-distilled oregano hydrolate acquired as by-products of essential oils purification were screened for their in vitro antimicrobial activity against three Salmonella Typhimurium strains (4/74, FS8, FS115) at 4 and 37 °C. Susceptibility to the extracts was mainly plant- and temperature-dependent, though strain dependent effects were also observed. Industrial oregano hydrolate eliminated strains immediately after inoculation, exhibiting the highest antimicrobial potential. Hydro-distilled extracts eliminated/reduced Salmonella levels during incubation at 4 °C. At 37 °C, oregano, centrifuged oregano, thyme, calendula and basil were bactericidal while spearmint, rosemary and corn silk bacteriostatic. A strain-dependent effect was observed for laurel. The individual or combined effect of marinades and edible coatings prepared of industrial hydrolate and hydro-distilled oregano extracts with or without oregano essential oil (OEO) was tested in pork meat at 4 °C inoculated with FS8 strain. Lower in situ activity was observed compared to in vitro assays. Marinades and edible coatings prepared of industrial oregano hydrolate + OEO were the most efficient in inhibiting pathogen. Marination in oregano extract and subsequent coating with either 50% oregano extract + OEO or water + OEO enhanced the performance of oregano extract. In conclusion, by-products of oregano essential oil purification may be promising alternative antimicrobials to pork meat stored under refrigeration when applied in the context of multiple hurdle approach.