Oxygen as a Possible Technological Adjuvant during the Crushing or the Malaxation Steps, or Both, for the Modulation of the Characteristics of Extra Virgin Olive Oil.

In commercial terms, Extra Virgin Olive Oil (EVOO) is considered an exceptional food with excellent sensory and nutritional quality due to its taste, odor, and bioactive compounds; as such, it is of great health interest. This quality can be affected by the oxidative degradation, both chemical and enzymatic (the activity of oxidative, endogenous enzymes from the polyphenol oxidase and peroxidase olive fruit type), of essential components during the extraction and conservation of EVOO. In the bibliography, oxygen reduction during the malaxation process and oil storage has been studied in different ways. However, research concerning oxygen reduction in the crushing of the olive fruit or the malaxation of the paste, or both, in the "real extraction condition" is scarce. Oxygen reduction has been compared to control conditions (the concentration of atmospheric oxygen (21%)). Batches of 200 kg of the olive fruit, 'Picual' cultivar, were used and the following treatments were applied: Control (21% O2 Mill-21% O2 Mixer), "IC-NM": Inerted crushing -Normal malaxation (6.25% O2 Mill-21% O2 Mixer), "NC-IM": Normal crushing-Inerted malaxation (21% O2 Mill-4.39% O2 Mixer) and "IC-IM": Inerted crushing -Inerted malaxation (5.5% O2 Mill-10.5% O2 Mixer). The parameters of commercial quality covered by regulation (free acidity, peroxide value and absorbency in ultra-violet (K232 and K270)) did not suffer any change concerning the control, and so the oils belong to the commercial category of "Extra Virgin Olive Oil". The phenolic compounds of the olives involved in the distinctive bitter and pungent taste, health properties, and oxidative stability are increased with the downsizing amounts of oxygen in the IC-NM, NC-IM, and IC-IM treatments with an average of 4, 10, and 20%, respectively. In contrast, the total amount of volatile compounds decreases by 10-20% in all oxygen reduction treatments. The volatile compounds arising from the lipoxygenase pathway, which are responsible for the green and fruity notes of EVOO, also decreased in concentration with the treatments by 15-20%. The results show how oxygen reduction in the milling and malaxation stages of olive fruit can modulate the content of phenols, volatile compounds, carotenoids, and chlorophyll pigments in the EVOO to avoid the degradation of the compound with sensorial and nutritional interest.

Endophytic Colonization by the Entomopathogenic Fungus Beauveria Bassiana Affects Plant Volatile Emissions in the Presence or Absence of Chewing and Sap-Sucking Insects.

Entomopathogenic fungi are gaining acceptance in Integrated Pest Management (IPM) systems as effective and environmental safety biological control agents to protect a great variety of crops against pest insects. Many of these insect-pathogenic fungi can establish themselves as endophytes and thereby may induce the plant immune system. The activation of plant defenses by the fungal endophytic colonization can have a direct impact on herbivores and plant pathogens. An integral component of many plant defense responses is also the release of volatile organic compounds, which may serve as an indirect defense by attracting the natural enemies of herbivores. Here we investigated the effect of endophytic colonization by the entomopathogenic fungus Beauveria bassiana on the volatile emission by melon and cotton plants, either unharmed or after being damaged by sap-sucking aphids or leaf chewing caterpillars. We found that when the plants are colonized by B. bassiana they emit a different blend of volatile compounds compared to uncolonized control plants. Some of the emitted compounds have been reported previously to be released in response to herbivory and have been implicated in natural enemy attraction. Several of the compounds are also known to have antimicrobial properties. Therefore, endophytic colonization by B. bassiana might help to not only direct control insect pests but also increase the resistance of plants against agronomically important pests and phytopathogens.

The ultrasound application does not affect to the thermal properties and chemical composition of virgin olive oils.

In this work, the effects of high power ultrasound treatment (40 kHz) on virgin olive oil (VOO) for different times (0, 15, 30 min) were studied, in order to verify if extent modifications in their chemical composition and thermal properties. The effects of the different ultrasound treatments on VOOs were determined considering the following parameters: quality index (free acidity, K232 and K270), lipid profile (fatty acids and triglycerides composition) minor components (phenols, tocopherols, pigments and volatiles) and thermal properties (crystallization and melting) by Differential Scanning Calorimetry (DSC). During the ultrasound treatments, bubbles growth was present in the VOO due to the phenomenon of cavitation and a slight increase of the temperature was observed. In general, the ultrasound treatments did not cause alterations on VOO parameters evaluated (oxidation state, lipid profile, minor components and thermal profiles). However, a slight decrease was observed in some volatile compounds.

How olive washing and storage affect fruit ethanol and virgin olive oil ethanol, ethyl esters and composition.

BACKGROUND: Olives are stored for a short time after harvesting pending processing in the oil mills. Furthermore, olives are often washed prior to fruit storage. In this work we study how washing and storage affect fruit ethanol content and the effect on virgin olive oil ethanol content and quality. RESULTS: Olive storage produced an increase in the fruit ethanol content, achieving values six times higher when storage was in silos. Washing the olives resulted in an increase in fruit ethanol content, although when washed olives were processed immediately no difference was found. The increase in fruit ethanol content during storage was reflected in higher oil ethanol concentration. Similarly, olive washing resulted in oils with higher ethanol concentration. Industrial conditions gave more important increases in oil ethanol content than that from olives processed by hand. For quality parameters all the olive oils were classified as 'extra virgin'. In general, oils showed a slight decrease in some sensory attributes. At industrial scale after 24 h storage oils were classified as 'virgin' because sensory defects were found. CONCLUSION: Olive storage should be avoided or reduced to less than 12 h; if possible, olives should not be washed before storage since this practice favors losses in sensory characteristics and the synthesis of ethanol, a precursor of ethyl esters. © 2020 Society of Chemical Industry.

Dietary Squalene Induces Cytochromes Cyp2b10 and Cyp2c55 Independently of Sex, Dose, and Diet in Several Mouse Models.

SCOPE: To investigate the effects of squalene, the main hydrocarbon present in extra virgin olive oil, on liver transcriptome in different animal models and to test the influence of sex on this action and its relationship with hepatic lipids. METHODS AND RESULTS: To this purpose, male C57BL/6J Apoe-deficient mice are fed a purified Western diet with or without squalene during 11 weeks and hepatic squalene content is assessed, so are hepatic lipids and lipid droplets. Hepatic transcriptomic changes are studied and confirmed by RT-qPCR. Dietary characteristics and influence of squalene doses are tested in Apoe-deficient on purified chow diets with or without squalene. These diets are also given to Apoa1 and wild-type mice on C57BL/6J background and to C57BL/6J xOla129 Apoe-deficient mice. Squalene supplementation increases its hepatic content without differences among sexes and hormonal status. The Cyp2b10 and Cyp2c55 gene expressions are significantly up-regulated by the squalene intake in all models, with independence of sex, sexual hormones, dietary fat content, genetic background and dose, and in Apoe-deficient mice consuming extra-virgin olive oil. CONCLUSION: Hepatic squalene increases the expression of these cytochromes and their changes in virgin olive oil diets may be due to their squalene content.

Effects of Endophytic Entomopathogenic Ascomycetes on the Life-History Traits of Aphis gossypii Glover and Its Interactions with Melon Plants.

Entomopathogenic fungi are sprayed commercially for aphid control in greenhouses. Recently, their ability to grow endophytically within plants was discovered, offering the opportunity for systemic biological control. Endophytic colonization of host plants could also influence life-table parameters and behavior of herbivores. We investigated lethal and pre-mortality effects of Beauveria bassiana and Metarhizium brunneum on Aphis gossypii; aphids either received inoculum while feeding on recently sprayed leaves (surface inoculum and endophytically-colonized) or while feeding on unsprayed but endophytically-colonized leaves. We used choice assays to identify any preferences for endophytically-colonized or control plants. Volatile emissions from endophytically-colonized plants and control plants were also compared. Aphid mortality rates ranged between 48.2 and 56.9 % on sprayed leaves, and between 37.7 and 50.0 on endophytically-colonized leaves. There was a significant effect of endophytic colonization on the rate of nymph production, but this did not result in an overall increase in the aphid population. Endophytic colonization did not influence host-plant selection even though there were qualitative and quantitative differences in the blend of volatiles released by endophytically-colonized and control plants. Although endophytic colonization did not change herbivore behavior, plants still benefit via indirect defense, resistance to plant pathogens or abiotic stress tolerance.

"Biosynthesis of volatile compounds by hydroperoxide lyase enzymatic activity during virgin olive oil extraction process".

The approach of this research was to describe the biochemical transformations of minor components of virgin olive oil with high impact on its sensory quality, specifically "volatile compounds" during its extraction process. For this purpose `Picual´, `Arbequina´ and `Hojiblanca´ cultivars were processed at three different harvesting times. Their volatile profiles and levels of enzymatic activity of "hydroperoxide lyase", responsible for the biosynthesis of C6 aldehydes related to "green and fruity notes", were monitored and identified in mesocarp, paste before kneading and paste after kneading based on the catalytic action of crude enzyme extracts. Both of them, volatiles and HPL, were analyzed by incubating with their corresponding substrates and reaction products formed were analyzed by Solid Phase Microextraction-Gas Chromatography and Mass Spectrometry. The results obtained in the present research can be useful in order to develop new markers biochemical whereby improved aroma quality in olive breeding programs or in the design of new protocol of VOO extraction.

Changes in Virgin Olive Oil Volatiles Caused by in Vitro Digestion.

Volatile compounds are responsible for some sensory characteristics of virgin olive oil (VOO); however, they have not been studied from a nutritional point view. In this work, the effect of the simulated digestion on VOO volatile compounds responsible for green flavor was studied, analyzing their changes through the three steps of an in vitro digestion model (mouth, stomach, and small intestine). Index of recovery and bioaccessibility were determined for the main volatiles of "Picual" VOO. At end of the duodenal step, higher recoveries of ethanol, pent-1-en-3-ol, ß-ocimene, and nonanal were observed. From the 10 volatile compounds analyzed, only six compounds were bioaccessible. The compounds with the highest bioaccessibility were pent-1-en-3-ol, nonanal, ß-ocimene, and ethanol. The results showed for the first time the recovery and bioaccessibility of several volatile compounds present in VOO.

Volatile Compound Profiling by HS-SPME/GC-MS-FID of a Core Olive Cultivar Collection as a Tool for Aroma Improvement of Virgin Olive Oil.

Virgin olive oil (VOO) is the only food product requiring official sensory analysis to be classified in commercial categories, in which the evaluation of the aroma plays a very important role. The selection of parents, with the aim of obtaining new cultivars with improved oil aroma, is of paramount importance in olive breeding programs. We have assessed the volatile fraction by headspace-solid-phase microextraction/gas chromatography-mass spectrometry-flame ionization detection (HS-SPME/GC-MS-FID) and the deduced aroma properties of VOO from a core set of olive cultivars (Core-36) which possesses most of the genetic diversity found in the World Olive Germplasm Collection (IFAPA Alameda del Obispo) located in Cordoba, Spain. The VOO volatile fractions of Core-36 cultivars display a high level of variability. It is mostly made of compounds produced from polyunsaturated fatty acids through the lipoxygenase pathway, which confirms to be a general characteristic of the olive species (Olea europaea L.). The main group of volatile compounds in the oils was six straight-chain carbon compounds derived from linolenic acid, some of them being the main contributors to the aroma of the olive oils according to their odor activity values (OAV). The high level of variability found for the volatile fraction of the oils from Core-36 and, therefore, for the aroma odor notes, suggest that this core set may be a very useful tool for the choice of optimal parents in olive breeding programs in order to raise new cultivars with improved VOO aroma.

Phenolic Compounds in Extra Virgin Olive Oil Stimulate Human Osteoblastic Cell Proliferation.

In this study, we aimed to clarify the effects of phenolic compounds and extracts from different extra virgin olive oil (EVOO) varieties obtained from fruits of different ripening stages on osteoblast cells (MG-63) proliferation. Cell proliferation was increased by hydroxytyrosol, luteolin, apigenin, p-coumaric, caffeic, and ferulic acids by approximately 11-16%, as compared with controls that were treated with one vehicle alone, while (+)-pinoresinol, oleuropein, sinapic, vanillic acid and derivative (vanillin) did not affect cell proliferation. All phenolic extracts stimulated MG-63 cell growth, and they induced higher cell proliferation rates than individual compounds. The most effective EVOO phenolic extracts were those obtained from the Picual variety, as they significantly increased cell proliferation by 18-22%. Conversely, Arbequina phenolic extracts increased cell proliferation by 9-13%. A decline in osteoblast proliferation was observed in oils obtained from olive fruits collected at the end of the harvest period, as their total phenolic content decreases at this late stage. Further research on the signaling pathways of olive oil phenolic compounds involved in the processes and their metabolism should be carried out to develop new interventions and adjuvant therapies using EVOO for bone health (i.e.osteoporosis) in adulthood and the elderly.

How 'ground-picked' olive fruits affect virgin olive oil ethanol content, ethyl esters and quality.

BACKGROUND: Olives dropped on the ground naturally sometimes are not separated from those fresh and healthy collected from the tree for harvest and processing. In this work we compared the quality, ethanol content and bioactive components of virgin olive oils from ground-picked olives, tree-picked fruits and their mixture. RESULTS: Ground-picked olives produced 'Lampante' virgin olive oils; these are of a lower quality category, because of important alterations in chemical and sensory characteristics. Ethyl esters showed the highest values, although under the regulated limit. The mixture of ground and tree-picked olives gave oils classified as 'virgin' because of sensory defects, although the quality parameters did not exceed the limits for the 'extra' category. Ethanol content showed a significant increase in the oils from ground- picked olives and their mixture with respect to those from tree-picked fruits. Furthermore, bioactive compounds showed a significant decrease as fruit quality was poorer. CONCLUSION: Ground-picked olives must be harvested and processed separately since they produce low-quality virgin olive oils with sensory defects and lower concentrations of bioactive compounds. The higher acidity and ethanol concentration observed in oils from ground-picked fruits or their mixture may help ethyl ester synthesis during storage. © 2015 Society of Chemical Industry.

Assessment of volatile compound profiles and the deduced sensory significance of virgin olive oils from the progeny of Picual×Arbequina cultivars.

Volatile compounds are responsible for most of the sensory qualities of virgin olive oil and they are synthesized when enzymes and substrates come together as olive fruit is crushed during the industrial process to obtain the oil. Here we have studied the variability among the major volatile compounds in virgin olive oil prepared from the progeny of a cross of Picual and Arbequina olive cultivars (Olea europaea L.). The volatile compounds were isolated by SPME, and analyzed by HRGC-MS and HRGC-FID. Most of the volatile compounds found in the progeny's oil are produced by the enzymes in the so-called lipoxygenase pathway, and they may be clustered into different groups according to their chain length and polyunsaturated fatty acid origin (linoleic and linolenic acids). In addition, a group of compounds derived from amino acid metabolism and two terpenes also contributed significantly to the volatile fraction, some of which had significant odor values in most of the genotypes evaluated. The volatile compound content of the progeny was very varied, widely transgressing the progenitor levels, suggesting that in breeding programs it might be more effective to consider a larger number of individuals within the same cross than using different crosses with fewer individuals. Multivariate analysis allowed genotypes with particularly interesting volatile compositions to be identified and their flavor quality deduced.

The Fatty Acid Composition of Virgin Olive Oil from Different Cultivars Is Determinant for Foam Cell Formation by Macrophages.

Although the beneficial role of Extra Virgin Olive Oil (EVOO) in the Mediterranean diet is well-known, its effects on health cannot be attributed solely to oleic acid. In addition to minor components, the presence of other fatty acids (FA), which depend largely on the cultivar among other factors, needs to be considered. The present study examined the effect of chylomicron remnant-like particles (CRLP) enriched in fatty acids of EVOO from 'Chetoui', 'Buidiego', 'Galega', 'Blanqueta', and 'Picual' cultivars on the foam cell formation by THP-1 macrophages. THP-1 cells were incubated with EVOO-CRLP for 24 h. Lipid accumulation in cells was measured by determining intracellular total triacylglycerol (TAG) concentration and FA composition. Intracellular TAG concentrations were higher in cells incubated with 'Chetoui' and 'Blanqueta' CRLP (0.33 ± 0.05 and 0.38 ± 0.07 µmol/mg of protein, respectively) than with 'Buidiego' and 'Picual' CRLP (0.20 ± 0.05 and 0.24 ± 0.06 µmol/mg of protein, respectively). In conclusion, linoleic acid-rich EVOO induced higher TAG incorporation into THP-1 macrophages compared to oleic acid-rich EVOO, the 18:1/18:2 ratio being consistently correlated with intracellular TAG accumulation. The results of this study demonstrated that the differences in EVOO-FA composition may have an important role in foam cell formation.

Ethanol in Olive Fruit. Changes during Ripening.

Ethanol is one of the precursors of ethyl esters, the virgin olive oil quality parameter for the "extra" category recently adopted by the European Union and International Olive Oil Council. Although ethyl ester content has great importance for virgin olive oil classification, the origin of ethanol is not clear. A possible source of ethanol may be the olive fruit itself while it remains on the tree. Variation of fruit ethanol content during ripening was studied for three different olive cultivars: 'Picual', 'Hojiblanca', and 'Arbequina'. Ethanol was measured in fruit homogenates by HS-SPME-GC-FID. The ethanol content varied between 0.56 and 58 mg/kg. 'Hojiblanca' fruits showed the highest ethanol concentration. For all of the cultivars, ethanol content of fruit increased during the ripening process, although a clear cultivar-dependent effect was observed because 'Hojiblanca' fruits showed the most significant raise. Therefore, results indicated that ethanol can be accumulated during fruit maturation on the olive tree.

First comprehensive characterization of volatile profile of north Moroccan olive oils: A geographic discriminant approach.

Herewith we have evaluated the variability of the composition in terms of volatile compounds of monovarietal "Picholine marocaine" olive oils and checked the possible influence of their geographical origin. For this purpose, 92 olive samples were collected during the harvesting period 2012/2013 from 7 north Moroccan regions, and the analysis of the volatile profiles of the obtained oils was performed by using headspace solid-phase microextraction coupled to gas chromatography with flame ionization and mass spectrometry detectors (HS-SPME/GC-FID-MS). A total of 40 volatile compounds belonging to different chemical classes were identified and quantified. Significant differences in the concentration levels of volatile constituents from oils of different geographical origins were found. Furthermore, for testing the ability of the identified volatile compounds for the geographical origin discrimination of the investigated oils, a stepwise linear discriminant analysis (s-LDA) was applied. Results revealed a very satisfactory classification of the studied oils according their geographic origin.

Variability of virgin olive oil phenolic compounds in a segregating progeny from a single cross in Olea europaea L. and sensory and nutritional quality implications.

Virgin olive oil phenolic compounds are responsible for its nutritional and sensory quality. The synthesis of phenolic compounds occurs when enzymes and substrates meet as olive fruit is crushed during the industrial process to obtain the oil. The genetic variability of the major phenolic compounds of virgin olive oil was studied in a progeny of the cross of Picual x Arbequina olive cultivars (Olea europaea L.). They belong to four different groups: compounds that included tyrosol or hydroxytyrosol in their molecules, lignans, flavonoids, and phenolic acids. Data of phenolics in the oils showed that the progeny displayed a large degree of variability, widely transgressing the genitor levels. This high variability can be of interest on breeding programs. Thus, multivariate analysis allowed to identify genotypes within the progeny particularly interesting in terms of phenolic composition and deduced organoleptic and nutritional quality. The present study has demonstrated that it is possible to obtain enough degree of variability with a single cross of olive cultivars for compounds related to the nutritional and organoleptic properties of virgin olive oil.

Factors limiting the synthesis of virgin olive oil volatile esters.

The aim of the present work was to establish the limiting factors affecting the biosynthesis of volatile esters present in virgin olive oil (VOO). Oil volatile fractions of the main Spanish olive cultivars, Arbequina and Picual, were analyzed. It was observed that acetate esters were the most abundant class of volatile esters in the oils, in concordance with the high content of acetyl-CoA found in olive fruit, and that the content of C6 alcohols is limited for the synthesis of volatile esters during the production of VOO. Thus, the increase of C6 alcohol availability during VOO production produced a significant increase of the corresponding ester in the oils in both cultivars at two different maturity stages. However, the increase of acetyl-CoA availability had no effect on the VOO volatile fraction. The low synthesis of these C6 alcohols seems not to be due to a shortage of precursors or cofactors for alcohol dehydrogenase (ADH) activity because their increase during VOO production had no effect on the C6 alcohol levels. The experimental findings are compatible with a deactivation of ADH activity during olive oil production in the cultivars under study. In this sense, a strong inhibition of olive ADH activity by compounds present in the different tissues of olive fruit has been observed.

Synthesis of volatile compounds of virgin olive oil is limited by the lipoxygenase activity load during the oil extraction process.

The aim of this work was to determine whether the lipoxygenase (LOX) activity is a limiting factor for the biosynthesis of virgin olive oil (VOO) volatile compounds during the oil extraction process. For this purpose, LOX activity load was modified during this process using exogenous LOX activity and specific LOX inhibitors on olive cultivars producing oils with different volatile profiles (Arbequina and Picual). Experimental data suggest that LOX activity is a limiting factor for the synthesis of the oil volatile fraction, this limitation being significantly higher in Picual cultivar than in Arbequina, in line with the lowest content of volatile compounds in the oils obtained from the former. Moreover, there is evidence that this limitation of LOX activity takes place mostly during the milling step in the process of olive oil extraction.

Oxygen concentration affects volatile compound biosynthesis during virgin olive oil production.

The effect of O 2 concentration on oil volatile compounds synthesized during the process to obtain virgin olive oil (VOO) was established. The study was carried out either on the whole process or within the main steps (milling and malaxation) of this process with two olive cultivars, Picual and Arbequina, at two ripening stages. Data show that O 2 control during milling has a negative impact on VOO volatile synthesis. This effect seems to depend on cultivar and on the ripening stage in cultivar Picual. Because most VOO volatiles are synthesized during olive fruit crushing at the milling step, O 2 control during malaxation seems to affect just slightly the volatile synthesis. The highest effect was observed when control of O 2 concentration was performed over the whole process. In this case, the content of volatile compounds of oils obtained from both cultivars and ripening stages showed quite similar trends.

Cultivar differences on nonesterified polyunsaturated fatty acid as a limiting factor for the biogenesis of virgin olive oil aroma.

The relationship between the content of nonesterified polyunsaturated fatty acids and the contents of oil aroma compounds that arise during the process to obtain virgin olive oil (VOO) was studied in two olive cultivars, Picual and Arbequina, producing oils with distinct aroma profiles and fatty acid compositions. Results suggest that the biosynthesis of VOO aroma compounds depends mainly on the availability of nonesterified polyunsaturated fatty acids, especially linolenic acid, during the process and then on the enzymatic activity load of the lipoxygenase/hydroperoxide lyase system. Both availability of substrates and enzymatic activity load seem to be cultivar-dependent.