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This research explores the health-promoting properties of the pectin–polyphenol complex extracted from alperujo, a by-product of olive oil production. This study investigates the chemical composition and antioxidant activity of the extracts, revealing their high antioxidant activity in vitro. Cell viability assays conducted on colon carcinoma cells (Caco-2) demonstrate the inhibitory effect of the extracts on cell proliferation. However, the extracts do not affect the viability of differentiated Caco-2 cells, suggesting a selective antiproliferative action. Additionally, the extracts reduce intracellular reactive oxygen species (ROS) and nitrite (NO) production in LPS-stimulated murine peritoneal macrophages. Furthermore, the extracts exhibit anti-inflammatory effects by downregulating the secretion of pro-inflammatory cytokines TNF-α, IL-1β, and IL-6 in these macrophages. These findings highlight the potential of pectin–polyphenol complexes as functional ingredients with significant health benefits, demonstrating antioxidant, antiproliferative, and anti-inflammatory properties. [ABSTRACT FROM AUTHOR]

Olive oil extraction by two-phase systems generates a by-product called "alperujo" which presents several difficulties for its valorization. The present work evaluated an industrial approach, based on the application of thermal treatments to alperujo followed by solid/liquid separation using standard two-phase olive oil mill equipment. Treatments consisted of the thermo-malaxation of alperujo at 70 °C for 45 or 90 min, with or without acid addition, followed by solid/liquid separation in an industrial decanter. The solid was characterized concerning subsequent use for composting, while total and hydrophilic phenolics were analyzed in liquid for their recovery. Additionally, a laboratory-scale trial to compare phenolic purification by ethylic acetate extraction with chromatographic procedures was also included. The static respiration index showed that solid fractions presented higher susceptibility to biodegradation processes than raw alperujo. The phenolic content of treated liquid fractions was higher than in raw alperujo. Total phenolics were maximum at the longest exposure time without acid addition, while hydrophilic phenolics were highest at the shortest exposure time in acidified samples. The use of non-ionic resins seemed attractive for obtaining highly concentrated phenolic fractions. The proposed thermal treatments can be applied in olive oil industries, allowing in situ pomace valorization and the recovery of phenolic-enriched liquid fractions. [ABSTRACT FROM AUTHOR]

Sperm cryopreservation is a fundamental tool for the conservation of avian genetic resources; however, avian spermatozoa are susceptible to this process. To cope with the high production of reactive oxygen species (ROS), the addition of exogenous antioxidants is beneficial. Pectoliv30 is a substance derived from alperujo, and in this study, its effect was analyzed on seminal quality after its addition to the cryopreservation extender of roosters at different concentrations. For this purpose, 16 Utrerana breed roosters were used, and seminal collection was performed in six replicates, creating a pool for each working day with ejaculates of quality. After cryopreservation, one sample per treatment and replicate was thawed, and several seminal quality parameters were evaluated. Statistical analysis revealed numerous correlations between these variables, both positive and negative according to the correlation matrix obtained. Furthermore, the chi-squared automatic interaction detection (CHAID) decision tree (DT) reported significant differences in the hypo-osmotic swelling test (HOST) variable between groups. Moreover, results for this parameter were more desirable at high concentrations of Pectoliv30. The application of this substance extracted from the by-product alperujo as an antioxidant allows the improvement of the post-thawing seminal quality in roosters and facilitates optimization of the cryopreservation process as a way to improve the conservation programs of different endangered poultry breeds. [ABSTRACT FROM AUTHOR]

Anaerobic digestion processes for biogas generation using alperujo have been the subject of intensive studies suggesting that phenolic compounds act as microbial inhibitors in anaerobic digestion. Pretreatments are needed to reduce the effects of phenolic compounds and improve biogas production, especially for dry anaerobic digestion. However, industrial‐scale implementation of these pretreatments is challenging, and it is unclear whether the improvement in biogas production justifies the energy expended on pretreatment. This study examines the energy analysis of dry anaerobic digestion and three alternative pretreatments: alkaline, oxidative and thermal. Results indicate that thermal pretreatment at 80°C with added water reduces phenolic compounds in alperujo by 35.4%. Meanwhile, pretreated with hydrogen peroxide in alkaline medium had the highest methane productivity (205 mL CH4/gVS). Even so, thermal pretreatment was only one with an energy return on investment greater than 1, signifying the necessity for energy analysis to ensure the viability of pretreatment processes. Highlights: Three pretreatments were evaluated to improve biogas production.Thermal pretreatment offers top phenolic compounds reduction in alperujo digestion.Oxidative pretreatment increased methane productivity by 36%.Dry anaerobic digestion shows promise for alperujo biogas production.Energy return on investment index determines thermal pretreatment efficiency and viability. [ABSTRACT FROM AUTHOR]

This research explores the health-promoting properties of the pectin–polyphenol complex extracted from alperujo, a by-product of olive oil production. This study investigates the chemical composition and antioxidant activity of the extracts, revealing their high antioxidant activity in vitro. Cell viability assays conducted on colon carcinoma cells (Caco-2) demonstrate the inhibitory effect of the extracts on cell proliferation. However, the extracts do not affect the viability of differentiated Caco-2 cells, suggesting a selective antiproliferative action. Additionally, the extracts reduce intracellular reactive oxygen species (ROS) and nitrite (NO) production in LPS-stimulated murine peritoneal macrophages. Furthermore, the extracts exhibit anti-inflammatory effects by downregulating the secretion of pro-inflammatory cytokines TNF-α, IL-1β, and IL-6 in these macrophages. These findings highlight the potential of pectin–polyphenol complexes as functional ingredients with significant health benefits, demonstrating antioxidant, antiproliferative, and anti-inflammatory properties.

Olive oil extraction by two-phase systems generates a by-product called “alperujo” which presents several difficulties for its valorization. The present work evaluated an industrial approach, based on the application of thermal treatments to alperujo followed by solid/liquid separation using standard two-phase olive oil mill equipment. Treatments consisted of the thermo-malaxation of alperujo at 70 °C for 45 or 90 min, with or without acid addition, followed by solid/liquid separation in an industrial decanter. The solid was characterized concerning subsequent use for composting, while total and hydrophilic phenolics were analyzed in liquid for their recovery. Additionally, a laboratory-scale trial to compare phenolic purification by ethylic acetate extraction with chromatographic procedures was also included. The static respiration index showed that solid fractions presented higher susceptibility to biodegradation processes than raw alperujo. The phenolic content of treated liquid fractions was higher than in raw alperujo. Total phenolics were maximum at the longest exposure time without acid addition, while hydrophilic phenolics were highest at the shortest exposure time in acidified samples. The use of non-ionic resins seemed attractive for obtaining highly concentrated phenolic fractions. The proposed thermal treatments can be applied in olive oil industries, allowing in situ pomace valorization and the recovery of phenolic-enriched liquid fractions.

Sperm cryopreservation is a fundamental tool for the conservation of avian genetic resources; however, avian spermatozoa are susceptible to this process. To cope with the high production of reactive oxygen species (ROS), the addition of exogenous antioxidants is beneficial. Pectoliv30 is a substance derived from alperujo, and in this study, its effect was analyzed on seminal quality after its addition to the cryopreservation extender of roosters at different concentrations. For this purpose, 16 Utrerana breed roosters were used, and seminal collection was performed in six replicates, creating a pool for each working day with ejaculates of quality. After cryopreservation, one sample per treatment and replicate was thawed, and several seminal quality parameters were evaluated. Statistical analysis revealed numerous correlations between these variables, both positive and negative according to the correlation matrix obtained. Furthermore, the chi-squared automatic interaction detection (CHAID) decision tree (DT) reported significant differences in the hypo-osmotic swelling test (HOST) variable between groups. Moreover, results for this parameter were more desirable at high concentrations of Pectoliv30. The application of this substance extracted from the by-product alperujo as an antioxidant allows the improvement of the post-thawing seminal quality in roosters and facilitates optimization of the cryopreservation process as a way to improve the conservation programs of different endangered poultry breeds.

In Spain and other Mediterranean countries, significant quantities of semi-solid olive mill waste are generated, which should be preferentially applied to agricultural soils to close nutrient cycles. However, two-phase olive mill waste (termed alperujo in Spanish) is known to pose risks to soil quality and plant production when applied to soil in large quantities. Alperujo has high contents of polyphenol substances, which can inhibit microbial growth and are also phytotoxic in nature. However, when applied in appropriate quantities and following specific methods, it is possible that the practice may not pose any risks, and this requires evaluation. As a waste management option, direct application of alperujo can supply plant nutrients and organic matter to degradation-prone Mediterranean soils. In order to validate this circular economy fertilization and soil protection strategy, an 18-month field experiment was undertaken, applying moderate quantities of alperujo on permanent crop groves throughout the Spanish region of Valencia. Eleven experimental parcels with permanent crops managed by farmers were identified to test two scenarios: a single application of 10 t/ha, and a second application of 10 t/ha after 10 months. Soil chemical parameters were assessed at 0, 6, 10, and 18 months. Soil organic carbon, total Kjeldahl nitrogen, pH, electrical conductivity, nitrates, and polyphenol contents were modified by alperujo application, but these effects were highly transient in nature, with generally no lasting effects after 4–6 months for either application scenario. Also, qualitative evaluations carried out by farmers revealed few effects, although some reductions in erosive processes and improvements in plant vigor were noted. As such, based on the measured parameters, it is concluded that the direct soil application of alperujo olive mill wastes at low application rates did not lead to any lasting detrimental effects on soil quality or compromise the productivity of permanent crops in this Mediterranean region. [ABSTRACT FROM AUTHOR]

Simple Summary: Olive oil production is the main economic interest in olive orchards, but this industrial process generates large amounts of a by-product called 'alperujo', which is very negative to the environment. When composted, it generates a product useful for fertilizing many crops, and the objective of the work was to evaluate its impact on the ant community in two diverse types of olive crop management: a superintensive grove (no-tillage, with a cover crop, irrigated) and a traditional grove (tillage, no cover crop, rain-fed). The main conclusion was that the addition of compost did not alter the presence of ants and the composition of the species in each grove, but the type of management could have influenced the abundance of ants (much more abundant in the superintensive than in the traditional grove) and, secondarily, the composition of the species. Industrial production of olive oil generates large amounts of solid waste called 'alperujo'. Its compost can fertilize many crops, especially olives. Furthermore, superintensive orchards are increasing their surface globally due to higher production and savings in different costs. Ants are considered an important part of the arthropod community in olive orchards and could even play a significant role in pest control. The tree canopy and ground were sampled to compare the ant assemblage in plots fertilized with compost and mineral products in two groves with different types of crop management (superintensive and traditional) over two years. The numbers of ants in both types of fertilization in each grove were not statistically different (p > 0.05), indicating that the type of fertilization did not have a significant impact on its populations in the ground or in the canopy, but the number of individuals was significantly higher in the superintensive grove than in the traditional grove (both in the ground and in the canopy, p < 0.01). The most frequent species in the ground were Pheidole pallidula, Plagiolepis smitzii and Aphaenogaster senilis (superintensive grove) and Pheidole pallidula, Tetramorium gr semilaeve, Plagiolepis pygmaea, and Tapinoma nigerrimum (traditional grove). In the canopy, the most frequent species were Plagiolepis spp. in both groves. Differences in ant densities and species between the groves could be due to the different management, especially of the soil, but it must be confirmed using more replicas and longer periods of study. [ABSTRACT FROM AUTHOR]

The objective of this study is to assess the inhibitory effects of an aqueous extract from olive oil mill waste (alperujo) on the growth of a lactic acid bacteria (LAB) cocktail consisting of various strains of Lactiplantibacillus pentosus and Lactiplantibacillus plantarum species. For this purpose, response surface methodology was employed using two independent variables (pH levels 3.5–5.55; hydroxytyrosol concentration ranging from 0.93–2990 ppm). The response variable was the average inhibition per treatment on the LAB cocktail (expressed as a percentage). The developed model identified significant terms, including the linear effect of hydroxytyrosol and pH, their interaction, and the quadratic effect of pH. Maximum inhibition of the LAB cocktail was observed at progressively higher concentrations of hydroxytyrosol and lower pH values. Therefore, complete inhibition of LAB in the synthetic culture medium could only be achieved for concentrations of 2984 ppm hydroxytyrosol at a pH of 3.95. These findings suggest that extracts derived from "alperujo" could be utilized as a natural preservative in acidified foods with a bitter flavor and antioxidant requirements. [ABSTRACT FROM AUTHOR]

Hydrophilic phenols are the main bioactive compounds in alperujo. Among them, 3,4-Dihydroxyphenylglycol (DHPG), Hydroxytyrosol (HT) and Tyrosol (Ty), are the most relevant and deeply studied. These compounds exhibit high antioxidant capacity and a wide range of health benefits as well as technologically promising properties. Given that, their recovery represents an attractive opportunity to valorize this by-product. In this work low thermal treatments were applied to alperujo in order to obtain phenol-enriched liquid fractions. Optimization assays combining different levels of temperature (30 to 90 ºC), time (60 to 180 min) and water content (70 to 90%), followed by response surface methodologies were performed. The results indicated that by applying optimal conditions, is possible to obtain theoretical yields of Total phenols, DHPG, HT and Ty of 2.4, 957.8, 3.4 and 6.4 times greater, respectively, than raw dry alperujo. Interestingly, all the evaluated conditions can be reproduced with low investment in a standard olive oil industry.

Hydrophilic phenols are the main bioactive compounds in alperujo. Among them, 3,4-Dihydroxyphenylglycol (DHPG), Hydroxytyrosol (HT) and Tyrosol (Ty), are the most relevant and deeply studied. These compounds exhibit high antioxidant capacity and a wide range of health benefits as well as technologically promising properties. Given that, their recovery represents an attractive opportunity to valorize this by-product. In this work low thermal treatments were applied to alperujo in order to obtain phenol-enriched liquid fractions. Optimization assays combining different levels of temperature (30 to 90 ºC), time (60 to 180 min) and water content (70 to 90%), followed by response surface methodologies were performed. The results indicated that by applying optimal conditions, is possible to obtain theoretical yields of Total phenols, DHPG, HT and Ty of 2.4, 957.8, 3.4 and 6.4 times greater, respectively, than raw dry alperujo. Interestingly, all the evaluated conditions can be reproduced with low investment in a standard olive oil industry. [ABSTRACT FROM AUTHOR]

In Spain and other Mediterranean countries, significant quantities of semi-solid olive mill waste are generated, which should be preferentially applied to agricultural soils to close nutrient cycles. However, two-phase olive mill waste (termed alperujo in Spanish) is known to pose risks to soil quality and plant production when applied to soil in large quantities. Alperujo has high contents of polyphenol substances, which can inhibit microbial growth and are also phytotoxic in nature. However, when applied in appropriate quantities and following specific methods, it is possible that the practice may not pose any risks, and this requires evaluation. As a waste management option, direct application of alperujo can supply plant nutrients and organic matter to degradation-prone Mediterranean soils. In order to validate this circular economy fertilization and soil protection strategy, an 18-month field experiment was undertaken, applying moderate quantities of alperujo on permanent crop groves throughout the Spanish region of Valencia. Eleven experimental parcels with permanent crops managed by farmers were identified to test two scenarios: a single application of 10 t/ha, and a second application of 10 t/ha after 10 months. Soil chemical parameters were assessed at 0, 6, 10, and 18 months. Soil organic carbon, total Kjeldahl nitrogen, pH, electrical conductivity, nitrates, and polyphenol contents were modified by alperujo application, but these effects were highly transient in nature, with generally no lasting effects after 4–6 months for either application scenario. Also, qualitative evaluations carried out by farmers revealed few effects, although some reductions in erosive processes and improvements in plant vigor were noted. As such, based on the measured parameters, it is concluded that the direct soil application of alperujo olive mill wastes at low application rates did not lead to any lasting detrimental effects on soil quality or compromise the productivity of permanent crops in this Mediterranean region.

Industrial production of olive oil generates large amounts of solid waste called ‘alperujo’. Its compost can fertilize many crops, especially olives. Furthermore, superintensive orchards are increasing their surface globally due to higher production and savings in different costs. Ants are considered an important part of the arthropod community in olive orchards and could even play a significant role in pest control. The tree canopy and ground were sampled to compare the ant assemblage in plots fertilized with compost and mineral products in two groves with different types of crop management (superintensive and traditional) over two years. The numbers of ants in both types of fertilization in each grove were not statistically different (p > 0.05), indicating that the type of fertilization did not have a significant impact on its populations in the ground or in the canopy, but the number of individuals was significantly higher in the superintensive grove than in the traditional grove (both in the ground and in the canopy, p < 0.01). The most frequent species in the ground were Pheidole pallidula, Plagiolepis smitzii and Aphaenogaster senilis (superintensive grove) and Pheidole pallidula, Tetramorium gr semilaeve, Plagiolepis pygmaea, and Tapinoma nigerrimum (traditional grove). In the canopy, the most frequent species were Plagiolepis spp. in both groves. Differences in ant densities and species between the groves could be due to the different management, especially of the soil, but it must be confirmed using more replicas and longer periods of study.

Olive oil production using the two-phase extraction system generates large amounts of biomass named alperujo , which has high percentage of water, slightly acidic pH, and high concentration of salts, organic matter, and phenolic compounds. Due to the presence of these components, alperujo is a significant source of environmental pollution, hindering the disposal of this waste. Nevertheless, some phenolic compounds such as tyrosol and hydroxytyrosol, have high nutraceutical and antioxidant power and could have potential applications in the food, pharmaceutical and cosmetic industries. This work aimed to evaluate the ability of different alperujo-derived media, with different levels of phenolic compounds, to sustain the growth and carotenoid production by yeasts from Rhodotorula genera (R. mucilaginosa LPSC 638 , R. glutinis Y54, and R. graminis Y52), as well as to prospect the biotechnological production of carotenoids in these media. The yeasts grew and produced carotenoids in all media, although growth was better in dephenolized media. The highest amount of carotenoids (43.0 ± 3.1 mg/g dry yeast) was obtained with R. glutinis Y54 grown in a thermally treated and dephenolized medium. Our study represents a novel approach to valorize alperujo with a dual benefit, the recovery of valuable compounds, i. e. phenolic compounds, and the bioproduction of new ones, i.e. carotenoids. [Display omitted] • Alperujo-derived mediums are suitable for bioprocess. • Dephenolization improves Rhodotorula growth and carotenoid production. • Rhodothorula growth reduces alperujo-derived mediums phytotoxicity. • Using Rhodotorula strains isolated from olive brines improves the whole process. • Dephenolization and use of endogenous strains gave higher carotenoid production. [ABSTRACT FROM AUTHOR]

Abstract The “alperujo” is a waste from the olive oil industry with great potential for valorization. It has a high organic load, with the presence of valuable compounds such as biophenols and sugars. The use of this waste can be thought of as a biorefinery from which different compounds of high added value can be obtained, whether they are present in the “alperujo” such as biophenols or can be generated from the “alperujo”. Therefore, the production of carotenoids by Rhodotorula mucilaginosa was evaluated using the liquid fraction of ‘alperujo’ (Alperujo Water, AW) or an aqueous extract (AE) of “alperujo” at different concentrations (5, 10, 20 and 30% w/V) as substrates. The AEs had an acidic pH, a total sugar concentration ranging from 1.6 to 7.6 g/L, a polyphenols content from 0.4 to 2.9 g/L and a significant amount of proteins (0.5–3 g/L). AW is similar in composition as 30% AE, but with a higher amount of total sugars. Rh. mucilaginosa was able to grow at the different mediums with consumption of glucose and fructose, a reduction in protein content and alkalinization of the medium. Maximum total carotenoid production (7.3 ± 0.6 mg/L) was achieved at AW, while the specific production was higher when the yeast grew at AW or at 30% AE (0.78 ± 0.06 and 0.73 ± 0.10 mg/g of biomass, respectively). Torulene and torularhodin were the main carotenoids produced. Polyphenol content did not change; thus, it is still possible to recover these compounds after producing carotenoids. These results demonstrate the feasibility of using alperujo-based mediums as cheap substrates to produce torularhodin and torulene and to include this bioprocess as a step in an integral approach for alperujo valorization.

The liquid fraction from the two-phase extraction process in the olive industry (alperujo), is a waste that contains lignocellulosic organic matter and phenolic compounds, difficult to treat by conventional biological methods. Lignocellulosic enzymes from white-rot fungi can be an interesting solution to break down these recalcitrant compounds and advance the treatment of that waste. In the present work the ability of Phanerochaete chrysosporium to degrade the abovementioned liquid waste (AL) was studied. Experiments were carried out at 26 °C within the optimal pH range 4–6 for 10 days and with and without the addition of glucose, measuring the evolution of COD, BOD5, biodegradability index, reducing sugars, total phenolic compounds, and colour. The results obtained in this study revealed the interest of Phanerochaete chrysosporium for an economical and eco-friendly treatment of alperujo, achieving COD and colour removals around 60%, and 32% of total phenolic compounds degradation, regardless of glucose addition. [ABSTRACT FROM AUTHOR]

Bladder cancer (BC) is one of the most common human cancers. There is an interest in controlling and treating BC and other types of cancer via the use of natural substances and/or combination chemotherapy. Modified forms of pectin have been reported to possess anticancer bioactivity related to the interaction of galactosyl, a main component of pectin, with galectin-3, a carbohydrate-binding protein that is overexpressed on many types of cancer cells. In this study, the antiproliferative effect on BC of novel modified pectins extracted from olives was evaluated. Pectoliv extracts, with high polyphenol content associated to polysaccharides rich in pectin, exhibited an important antiproliferative capacity in vitro against four human BC cells lines, RT112, T24, J82, and SCaBER. Pectoliv treatment reduced the expression of galectin-1 and galectin-3 and significantly inhibited the agglutination of erythrocytes. Thus, Pectoliv may have the potential for development as a novel galectin-3 inhibitor. [ABSTRACT FROM AUTHOR]

Thermal treatments are the latest developments in the olive oil biorefinery industry to extract bioactive compounds from its by-products, mainly the alperujo. To reach these goals and reduce energy consumption the utilization of deep eutectic solvents has been studied. An initial screening led to define a eutectic mix of choline chloride, glycolic and oxalic acid (DES9) as one of the adequate solvents to increase the solubilization of phenols and sugars, with a temperature reduction from 180 °C to 120 °C. DES9 increased the concentration of acid sugars by six times and the concentration of hydroxytyrosol by 30 times, to up to 85.81 mg/g of dry matter. The activity of DES9 is not due to the activity of each component separately or to the mixture of the two acids, but to the eutectic mixture of all of them. In the future, these solvents could improve the extraction, stability and bioavailability of bioactive compounds and the biorefinery of alperujo. [ABSTRACT FROM AUTHOR]

The aim of this research was to investigate the interaction of phenols and pectic polysaccharides during the olive oil extraction process. For this, pectin was extracted from fresh olive fruits and compared to the pectin isolated from the paste resulting from the extraction of the olive oil after milling with malaxation at 30 °C/30 min and subsequent centrifugation of the olive paste from the same lot of olive fruits in a system called ABENCOR (AB). The results indicate that these interactions were enhanced during the olive oil extraction process. In addition, the resulting AB extracts exhibited high antioxidant activity (ORAC) and strong antiproliferative activity in vitro against colon carcinoma Caco-2 cell lines compared to olive fruit extracts. The polyphenols associated mainly with the acidic pectin substance, with a higher content in AB extracts, seem to be responsible for these activities, and appear to maintain their activities in part after complexation. However, even in olive fruit extracts with smaller amounts of phenols in their compositions, pectic polysaccharides may also be involved in antioxidant and antiproliferative activities.

The liquid fraction from the two-phase extraction process in the olive industry (alperujo), is a waste that contains lignocellulosic organic matter and phenolic compounds, difficult to treat by conventional biological methods. Lignocellulosic enzymes from white-rot fungi can be an interesting solution to break down these recalcitrant compounds and advance the treatment of that waste. In the present work the ability of Phanerochaete chrysosporium to degrade the abovementioned liquid waste (AL) was studied. Experiments were carried out at 26 °C within the optimal pH range 4–6 for 10 days and with and without the addition of glucose, measuring the evolution of COD, BOD5, biodegradability index, reducing sugars, total phenolic compounds, and colour. The results obtained in this study revealed the interest of Phanerochaete chrysosporium for an economical and eco-friendly treatment of alperujo, achieving COD and colour removals around 60%, and 32% of total phenolic compounds degradation, regardless of glucose addition.

Extracts rich in pectic polysaccharides associated to polyphenols were obtained from alperujo, the by-product of olive oil extraction, by different thermal treatments and subsequent chemical treatments. The resulting “Pectoliv” extracts exhibited strong antioxidant activities and a high antiproliferative capacities in vitro against colon carcinoma Caco-2 and leukemia monocytic THP-1 cell lines. Bleaching of the extracts with sodium chlorite removed the phenolic compounds and significantly reduced antiproliferative and antioxidant activities, confirming that these were due mainly to the associated polyphenols present in Pectoliv. However, even after bleaching, these activities remained very similar to that of a commercial citrus modified pectin obtained by chemical treatment from citrus pectin, which has been implicated in cancer treatment and prevention. Thus Pectolivs have a high potential as antiproliferative or antioxidant agents.

"Alperujos" of Arbequina olives harvested in 2010 and 2011 and of Nevadillo olives in 2012 were evaluated in order to reduce their pollutant load by recovering the phenols with antioxidant properties. First, three successive extractions with water (4 °C, 120 min) were carried out to evaluate by spectrophotometry the total phenol contents, which significantly varied with cultivar and harvest year. The combined aqueous extracts of "alperujo" (combined-AEA) showed high pollutant loads with volatile solids to total solids ratios close to 1 suggesting high organic material contents and very low total phenols to chemical oxygen demand ratios. Second, phenols were recovered from the combined-AEA using ethyl acetate. By experimental design, the following conditions were found to maximize phenol recovery: temperature = 20 °C, time = 120 min, pH of aqueous extract = 2.00 and solvent/extract ratio (v/v) = 3:1. Up to 41.6% of the phenols were recovered from the combined-AEA after three successive extractions using recycled solvent. [ABSTRACT FROM AUTHOR]

The development of thermal treatments is helping to optimize the management of waste generated by the two-phase olive oil extraction system. A new treatment reactor, based on the steam explosion system, has been scaled up to the industrial level and offers several physical and chemical advantages. The reactor has been adapted to work into the pomace oil extractors. Chemical advantages include the efficient solubilization of simple phenols, like hydroxytyrosol, and sugars in the liquid fraction, and a significant reduction of humidity and toxicity of the residual solid fraction, in which cellulose and oil are concentrated, making the solid residual waste more suitable for subsequent bioprocess applications. All of these parameters will lead to the widespread use of steam treatment in the near future to improve the management and obtain the maximum exploitation of this byproduct. • Thermal treatment for alperujo is helping to optimize it industrial management. • A new treatment reactor has been scaled up to the industrial level. • The reactor offers several physical and chemical advantages. • The advantages will lead to the widespread use of the treatment in the near future. [ABSTRACT FROM AUTHOR]

The current management of alperujo as the main solid by-product from the two-phase olive oil extraction system has led to the appearance of a new liquid effluent that until now was treated together with the alperujo itself. The composition and antioxidant properties of its bioactive components at different depths of the pond were studied using colorimetric and HPLC with UV and MS detectors, DPPH, reducing power and rancimat. The concentration of suspended solids varied between 1.71 and 8.49 g/L, total fat was between 0.74 and 1.47 g/L, and total phenols were found between 3.74 and 4.11 g/L, which included hydroxytyrosol, 3,4-dihydroxyphenylglycol and tyrosol as the main phenols. Two types of extracts were obtained through two industrial systems with ethyl acetate and by chromotography, with an average content in total sugars of 2.1% and 3.16%, total phenols of 17.9% and 28.6% and hydroxytyrosol of 51.5 and 79.0 mg/g of extract, respectively. The activity presented by the chromatographic extract was higher in terms of free radical sequestering capacity, reducing power and the inhibition of lipid oxidation. Obtaining bioactive extracts would improve the formulation of food with natural components and at the same time would be the first step in a biorefinery to improve the management of the new effluent.

Pomace is an oil mill byproduct and the raw material for extracting olive pomace oil. The pomace is stored several months in ponds until it is processed. Tests have been carried out in stored olive pastes, malaxated with and without different coadjuvants. These pastes have been equated to the oil mill pomace. Coadjuvants increase the water pH in 0.8 units. 89.5% of the initial oil is recovered from pastes without coadjuvant; 91.1% from pastes with calcium carbonate and 80.5% from pastes with talc. The free acidity is 1.9%, 2.6% and 2.1% respectively. The maximum amount of refined pomace oil is obtained when calcium carbonate is used and the minimum when talc is used. Moisten the adjuvants affects talc and calcium carbonate differently, although both of them easily releases the original micro-powder. Moisture does not affect the technological action of the coadjuvants or modify their physical characteristics. [ABSTRACT FROM AUTHOR]

The two‐phase centrifugation system of olive oil extraction produces huge amounts of olive pomace called alperujo that could be a good source of bioactive substances. However, alperujo is currently stored in open air ponds before it is dried and submitted for extraction of the residual oil. The aim of this work is to study the evolution of phenolic and triterpenic acids in alperujo during its storage and extraction processes on an industrial scale. Alperujo is stored in large open air ponds, dried and the residual oil is extracted with hexane over a course of a year for two consecutive olive seasons.The storage of alperujo did not cause great changes in the content of these substances in the paste although the pomace olive oil is enriched in triterpenic compounds, mainly oleanolic acid. By contrast, the drying and extraction steps gives rise to a great reduction in the concentration of phenolic compounds. Likewise, triterpenic acids are very stable during the extraction process, and an enrichment of them is detected in the extracted alperujo that can reach up to 7–8 g kg−1 in these substances, particularly maslinic acid. Practical Applications: There are many studies, methods, and patents to extract bioactive substances from olive by‐products, particularly from the fresh olive pomace called alperujo. However, this study is the first to explore the stored and extracted alperujo as a good source of phenolic and triterpenic substances. Specifically, triterpenic acids were very stable during the storage and drying steps and extracted alperujo was highly enriched in them. This research will contribute to the valorization of olive by‐products. Extracted alperujo is a very good source of triterpenic acids. [ABSTRACT FROM AUTHOR]

Olive mill by-products are effluents generated during olive oil production process. The two-phase centrifugation system produces a semi-solid olive pomace called "alperujo." This by-product is a combination of liquid and solid wastes derived from the three-phase manufacturing process. A direct and fast analytical method by ultra-high-pressure liquid chromatography-DAD coupled with ESI/MS-MS has been developed for the profiling of phenolic compounds. Thirtyfive metabolites belonging to phenyl alcohols, secoiridoids, flavonoids, and iridoids were identified as the main constituents of alperujo in 12 min, including p-coumaroyl aldarate and a verbascoside derivative found for the first time in alperujo and a new ligstroside derivative. Six quantitatively significant components were determined at concentrations ranging from 17.7 mg/L for p-coumaric acid to 370.7 mg/L for hydroxytyrosol. Our data confirm that alperujo is an interesting source of phenolic compounds that could be extracted for use as nutraceuticals. [ABSTRACT FROM AUTHOR]

Onion is an allogamous species with protandry, as anthers release pollen before the stigma is receptive. Thus, insect pollination is needed to produce seeds. When male sterile lines are used, an efficient pollinator is important to ensure pollen transfer to the stigma. Therefore, increasing seed yield depends on increasing bee activity. Seed yield is variable in hybrid lines and lower than open-pollinated varieties, with a decrease of up to 60%. Bee attractants sometimes are used to influence worker bees towards flowers for pollination. The objective of this work is to use a food industry waste, such as olive pomace, as bee attractant upon application of the water extract on onion umbels. The effectiveness of this by-product in enhancing pollination efficiency was evaluated and compared to untreated controls. Male-sterile lines, as well as a fertile line, were used in commercial fields and environmental variables were registered. Pollination effectiveness was measured by seed yield. Olive pomace water extracts (OPWE), rich in bioactive compounds, from local industries were evaluated. Some compounds present in the OPWE were analyzed, identified and quantified. The OPWEs were applied during flowering, at different times and correlated with seed yield and the frequency of bee visits. After 3 years of study, we can assert that the application of OPWE enhances honeybee pollination efficiency, and as consequence improves onion seed yield. [Display omitted] • The application of olive pomace water extract enhances onion seed yield. • Revaluation of olive pomace to improve onion seed yield. • A waste from food industry, such as olive pomace, was used as bee attractant. • Olive pomace water extract is rich in bioactive compounds. • Male as well male sterile lines showed an improvement in seed yield. [ABSTRACT FROM AUTHOR]

The application of a novel industrial process based on the hydrothermal treatment of olive oil waste (alperujo) led to a final liquid phase that contained a high concentration of simple phenolic compounds. In this study the effect of pH on phenol extraction with ethyl acetate from the aqueous phase of hydrothermally treated alperujo at 160 °C/60 min (without modification, pH 4.5, and adjusted to pH 2.5) was evaluated, beside the increase of hydroxytyrosol during the storage. The variation of the concentration of phenolic compounds in each extract was analyzed by HPLC. The phenolic extract obtained at pH 4.5 presented a higher proportion of total and individual phenols and better antioxidant capacity in vitro than the extract obtained at pH 2.5. The use of lower pH values enhances the concentration of hydroxytyrosol in the liquid diminishing the storage times. [ABSTRACT FROM AUTHOR]

The effect of a microwave (MW) pre-treatment on two-phase olive mill solid residue (OMSR) or alperujo with a view to enhancing its anaerobic digestibility was studied. The MW pre-treatment was carried out at a power of 800 W and at a targeted temperature of 50 °C using different heating rates and holding times. The following specific energies were applied: 4377 kJ·kg TS-1 (MW1), 4830 kJ·kg TS-1 (MW2), 7170 kJ·kg TS-1 (MW3) and 7660 kJ·kg TS-1 (MW4). The maximum methane yield, 395±1 mL CH4·g VSadded-1, was obtained for MW4. The effect of the pre-treatment on the kinetics of the process was also studied. The methane production curves generated during the batch tests showed a first exponential stage and a second sigmoidal stage for all the cases studied. In the first stage, the kinetic constant for the pre-treatment MW1 was 54.8% higher than that obtained for untreated OMSR.

El objetivo de este trabajo ha sido estu­diar el efecto de la aplicación de alperujo (residuo de almazara) sobre la persisten­cia y el movimiento del herbicida diuron [3-(3,4-diclorofenil)-1,1-dimetilurea] en un olivar en Mengíbar (Jaén, España). La parcela se dividió en 2 subparcelas, un en la que no se aplicó enmienda al suelo y otra donde se aplicó alperujo (18000 kg ha-1). Tras la aplicación del herbicida a una dosis de 2 kg/ha se tomaron muestras de suelo por triplicado en cada subparce­la, a distintas profundidades y distintos días desde la aplicación de diuron. Des­pués de secar, limpiar las muestras de sue-lo y tamizarlas, se procedió a la extrac­ción de diuron con metanol y se analizó por HPLC. Se observó una mayor canti­dad de diurón en el suelo enmendado que en el suelo no enmendado en todas las muestras. Sin embargo, la movilidad de diuron no aumentó con el tiempo en el suelo enmendado.The aim of this work was to study the ef­fect of "alperujo" (olive mill waste) on the persistence and the mobility of the herbicide diuron [3-(3,4-dichlorophenyl)-1,1-dime­thylurea] in an olive grove from Mengíbar (Jaén, Spain). The plot was divided in two subplots, one where no amendement was applied and another one where "alperujo" was applied to the soil (18.000 kg ha-1). Af­ter herbicide application at a rate of 2 kg/ha, three replicates of soil were sampled at each subplot, at different depths and different days after diuron application. After drying, cleaning and sieving samples diuron was extracted with methanol and the extracts were analyzed by HPLC. Higher amounts of diuron were recovered from amended soil than from non amended soil in every sam­ple. However, diuron mobility in amen-ded soil did not increase with time.

The effect of a microwave (MW) pre-treatment on two-phase olive mill solid residue (OMSR) or alperujo with a view to enhancing its anaerobic digestibility was studied. The MW pre-treatment was carried out at a power of 800 W and at a targeted temperature of 50 °C using different heating rates and holding times. The following specific energies were applied: 4377 kJ·kg TS−1 (MW1), 4830 kJ·kg TS−1 (MW2), 7170 kJ·kg TS−1 (MW3) and 7660 kJ·kg TS−1 (MW4). The maximum methane yield, 395±1 mL CH4·g VSadded−1, was obtained for MW4. The effect of the pre-treatment on the kinetics of the process was also studied. The methane production curves generated during the batch tests showed a first exponential stage and a second sigmoidal stage for all the cases studied. In the first stage, the kinetic constant for the pre-treatment MW1 was 54.8% higher than that obtained for untreated OMSR. [ABSTRACT FROM AUTHOR]

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Environmentally acceptable disposal of olive cultivation residues (e.g., olive prunings; olive pruning residues (OLPR)) and olive mill wastes is of paramount importance since they are generated in huge quantities within a short time. Moreover, olive mill wastewater (OMW) or sludge-like effluents ('alperujo'; two-phase olive mill waste (TPOMW)) are highly biotoxic. Hericium erinaceus is a white-rot fungus which produces choice edible mushrooms on substrates rich in lignocellulosics, and its suitability for the treatment of olive by-products was examined for the first time. Fungal growth resulted in a notable reduction of OMW's pollution parameters (i.e., 65 % decolorization, 47 % total phenolic reduction, and 52 % phytotoxicity decrease) and correlated with laccase and manganese peroxidase activities. Solid-state fermentation of various mixtures of OLPR, TPOMW, and beech sawdust (control) by H. erinaceus qualified OLPR in subsequent cultivation experiments, where it exhibited high mushroom yields and biological efficiency (31 %). Analyses of proximate composition and bioactive compound content revealed that mushrooms deriving from OLPR substrates showed significantly higher crude fat, total glucan, β-glucan, total phenolics, and ferric-reducing antioxidant potential values than the control. H. erinaceus demonstrated the potential to detoxify OMW and bioconvert OLPR into high-quality biomass, and hence, this fungus could be successfully exploited for the treatment of such by-products. [ABSTRACT FROM AUTHOR]

This study was carried out to determinate phenolic, flavonoid content, and antioxidant capacity in methanolic extract from three Alperujo varieties. Alperujo Barnea showed the highest concentration of phenols and flavonoid. The greater hydroxytyrosol content was obtained in the same extract (4.93 ± 0.37 µg/mg extract), whereas the greater tyrosol content (0.23 ± 0.012 µg/mg extract) was found in Arbequina extract. These results were correlated with the greatest radical scavenging and the highest inhibition of lipoperoxidation process observed in Barnea extract (IC50of 27.9 ± 1.04 µg/mL; IC5022.8 ± 3.5 µg/mL, respectively). In spite of differences, alperujo extracts exhibited notable antioxidant capacities. [ABSTRACT FROM AUTHOR]

In the olive fruits there is a large amount of bioactive compounds and substances of high interest. Many of them are known by owing health beneficial properties that contribute to protective effect of the virgin olive oil. During olive oil processing, most of them remain in the olive oil wastes. Although, olive-mill wastewater (OMWW) or “alpechin”, olive oil cake (OOC), and the new by-product, known as “alperujo” in Spain and generated by the two-phase extraction process, represent a major disposal and potentially severe pollution problem for the industry, they are also promising source of substances of high value. This review summarises the last knowledge on the utilisation of residual products, with more than 90 references including articles and patents, which are promising with regard to future application. All these investigations have been classified into two options, the recovery of valuable natural constituents and the bioconversion into useful products.

Olive mill waste (OMW) creates a major environmental problem due to the difficulty of further waste processing. In this work we present an approach to give OMW added value by using it for the production of biosurfactants. Two bacterial species, Pseudomonas aeruginosa and Bacillus subtilis, were grown with OMW as the sole carbon source. Glycerol and waste frying oil were used as comparative carbon sources. B. subtilis produced surfactin (a lipopeptide) at a maximum concentration of 3.12 mg/L with 2% w/v of OMW in the medium, dropping to 0.57 mg/L with 10% w/v of OMW. In contrast, P. aeruginosa produced 8.78 mg/L of rhamnolipid with 2% w/v OMW increasing to 191.46 mg/L with 10% w/v OMW. The use of solvent-extracted OMW reduced the biosurfactant production by 70.8% and 88.3% for B. subtilis and P. aeruginosa respectively. These results confirm that OMW is a potential substrate for biosurfactant production. [ABSTRACT FROM AUTHOR]

Purpose: This study explored the in vitro antioxidant and anti-platelet activities of hydroxytyrosol, hydroxytyrosol acetate, 3,4-dihydroxyphenylglycol and two phenolic olive extracts. These compounds and extracts were obtained from a new industrial process to hydrothermally treat the alperujo (160 °C/60 min), a by-product of olive oil extraction. Methods: The extracts and the purified compounds were obtained chromatographically using both ionic and adsorbent resins. The antioxidant activity was determined by measuring inhibition of human platelet aggregation and inhibition of lipid peroxidation in liver microsomes of vitamin E-deficient rats. Results: The positive effect of the extracts on the inhibition of platelet aggregation is showed, being higher in the case of hydroxytyrosol acetate up to 38 %, and for the first time, its synergist effect with hydroxytyrosol has been proved, obtaining more than double of inhibition. The phenolic extracts and the isolated phenols showed good results for inhibiting the lipid oxidation, up to 62 and 25 %, respectively. A synergistic effect occurred when the hydroxytyrosol acetate and the 3,4-dihydroxyphenylglycol were supplemented by hydroxytyrosol. Conclusion: These results suggest the extract and these compounds obtained from a novel industrial process could be natural alternatives for the prevention of diseases related to cardiovascular disorder or oxidative damage. [ABSTRACT FROM AUTHOR]

BACKGROUND Alperujo is the paste generated from the two-phase extraction system of olive oil. This wet pomace must be stored for several months and, during this period, the formation of 4-ethyphenol provokes a strong off-odour. The aim of this work was to identify the microorganisms able to produce this volatile phenol. RESULTS Yeast and bacterial strains were isolated from stored alperujo and tested for their ability to metabolize p-coumaric acid and form 4-ethylphenol. Among them, Lactobacillus pentosus was the microorganism that both in synthetic medium and alperujo gave rise to 4-ethylphenol formation. This microorganism did not grow in alperujo acidified to pH 2, thereby confirming that acidification as the best method to control odour emissions during alperujo storage. CONCLUSION Lactic acid bacteria, particularly Lactobacillus pentosus, can be responsible for the formation of the off-odour caused by 4-ethylphenol during the storage of alperujo. This odour can be prevented by acidifying the alperujo. © 2014 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

The reuse of agricultural waste is essential to mitigate environmental pollution problems. Consequently, this work focused on the valorization of the most extensive olive processing residue, alperujo , in terms of its ability to interfere with the virulence processes of Pseudomonas aeruginosa and Staphylococcus aureus contaminating food and reference strains. The activity of alperujo extracts of different polarities on virulence factors producing bacteria, and its relationship with antioxidant capacity and polyphenolic content was comparatively evaluated. The results indicated that the polar extracts significantly reduced the biofilm formed by both bacteria. They were also able to act against a previously formed biofilm by reducing its biomass and the cells' metabolism. Ethanolic extracts, rich in polyphenolic compounds, exhibited antioxidant capacity and Quorum sensing (QS) inhibitory activity against Chromobacterium violaceum. Additionally, they limited the spread of P. aeruginosa by restricting swarming and swimming movements. They also interfere in other QS controlled phenotypes like elastase and pyocyanin. The antipathogenic ability of the polar extracts could be mainly due to the synergistic effect of several phenolic compounds identified by LC–MS/MS analysis, such as luteolin, vanillic acid hexoside, oleuropein and derivates, quinic acid, apigenin, among others. As a result, this study contributes to the developing of sustainable strategies to revalorize a polluting waste into a value-added byproduct that could be used in the food and sanitary industries. • Alperujo extracts attenuate the biofilm formed by P. aeruginosa and S. aureus. • They disrupt a preformed biofilm and decrease its metabolic activity. • They limit the spread of P. aeruginosa by restricting swarming and swimming movements. • Alperujo polar extracts inhibit Quorum sensing , pyocyanin and elastase activity. • Ethanolic extracts rich in polyphenolic compounds exhibit antioxidant capacity. [ABSTRACT FROM AUTHOR]

The pectin fraction released from steam-treated alperujo at 160 °C for 30, 45, and 60 min was purified and characterized. Differences were observed in the composition, and physical and biological activities of the pectin extracts, depending on the length of the thermal treatment. The extracts presented a low molecular weight in the range of 2–40 KDa, a high content of neutral sugars, and a high percentage of acetylation. In comparison with commercial pectins, the extracts had low water and high oil holding capacities, a normal emulsifying activity and a similar emulsion stability to that of apple pectin. For the three pectin extracts studied, the in vitro analyses showed considerable bile-acid binding activity and a glucose retardation index similar to the values obtained for citrus pectins. Thus, the pectins isolated from olive oil waste have suitable physical and biological properties for commercial use. Pectin extraction from alperujo, a sizeable by-product of the olive oil manufacturing process, would facilitate the use and subsequent reduction of this environmentally damaging organic waste. [ABSTRACT FROM AUTHOR]

The use of poultry manure or goat/sheep manure in the co-composting of the two-phase olive-mill cake “alperujo” (ALP) with olive leaf (OL) is compared by studying organic-matter mineralization and humification processes during composting and the characteristics of the end products. For this, two different piles (P1 and P2) were prepared using ALP with OL mixed with poultry manure (PM) and goat/sheep manure (GSM), respectively, and composted by the turned windrow composting system. Throughout the composting process, a number of parameters were monitored, such as temperature, pH, electrical conductivity (EC), organic matter (OM), OM losses, total organic carbon (Corg), total nitrogen (Nt), Corg/Ntratio, and the germination index (GI). In both piles, the temperature exceeded 55 °C for more than 2 weeks, which ensured maximum pathogen reduction. Organic-matter losses followed a first-order kinetic equation in both piles. The final composts presented a stabilized OM and absence of phytotoxins, as observed in the evolution and final values of the Corg/Ntratio (Corg/Nt < 20) and the germination index (GI > 50 percent). Therefore, composting can be considered as an efficient treatment to recycle this type of waste, obtaining composts with suitable properties that can be safely used in agriculture. [ABSTRACT FROM PUBLISHER]

Olive oil is an integral ingredient of the “Mediterranean diet”. The olive oil industry generates large quantities of a by-product called “alperujo” (AL) during the two-phase centrifugation system developed in the early nineties. AL could be a potent exploitable source of natural phenolic antioxidants. Our results showed that AL and its distinctive phenols hydroxytyrosol, tyrosol and verbascoside were not genotoxic in the Somatic Mutation and Recombination Test (SMART) of Drosophila melanogaster and exerted antigenotoxic activity against DNA oxidative damage generated by hydrogen peroxide (H 2 O 2 ). Alperujo and hydroxytyrosol also exhibited notable antiproliferative and caspase 3-dependent proapoptotic effects toward the human tumoral cell line HL60. AL can provide a cheap and efficient source of chemopreventive phenolic compounds with strong antioxidant properties, becoming a promising and potent therapeutic drug in the future. [ABSTRACT FROM AUTHOR]