Lipoxygenase activity in olive (Olea europaea) fruit

The present work was designed to characterize lipoxygenase activity in olive fruit pulp, in order to determine its significance in the biosynthesis of virgin olive oil aroma. Lipoxygenase activity has been detected in particulate fractions of enzyme extracts from olive pulp subjected to differential centrifugation. The activity in different membrane fractions showed similar properties, with optimal pH in the range of 5.0–5.5 and a clear specificity for linolenic acid, which was oxidized at a rate double that of linoleic acid under the same reaction conditions. The enzyme preparations displayed very low activity with dilinoleoyl phosphatidylcholine, suggesting that olive lipoxygenase acts on nonesterified fatty acids. The enzyme showed regiospecificity for the Δ-13-position of both linoleic and linolenic acid, yielding 75–90% of Δ-13-fatty acid hydroperoxides. Olives showed the highest lipoxygenase activity about 15 wk after anthesis, with a steady decrease during the developmental and ripening periods. Olive lipoxygenase displayed properties that support its involvement in the biogenesis of six-carbon volatile aldehydes, which are major constituents of the aroma of virgin olive oil, during the process of oil extraction.     

Change in the natural ratio between chlorophylls and carotenoids in olive fruit during processing for virgin olive oil

Different varieties of olives from different sources, harvested in a similar ripeness state, have been characterized by their chlorophyll and carotenoid pigment profile and content. Pigment richness is inherent to the variety and enables great differences or similarities to be established between them. In all the fruits, ripening involves pigment loss, with the disappearance of chlorophylls always being slightly greater than that of carotenoids. However, independently of variety and the different pitment content of the fruits, the ratio between chlorophylls and carotenoids tends to remain more or less constant, within a range of 2.5 to 3.7 mg total chlorophyll/mg total carotenoid. Pigment loss caused by the extraction process is more marked for the chlorophyll fraction than for the carotenoids, changing the ratio in the oil to around one unit, whatever the variety of soruce fruit.     

Rapid evaluation of olive oil quality by NIR reflectance spectroscopy

NIR spectroscopy calibrations have been developed for a range of quality parameters in olive oil, including FFA, PV, polyphenol content, induction time, chlorophyll, and the major FA. A set of 216 olive oil samples from throughout the Australian olive-growing areas were used to provide a representative range of quality. The variation in the oils tested virtually covered the range of the chemical standard limits described by the International Olive Oil Council. A FOSS NIRSystems^® 6500 spectrophotometer with a liquid cell holder was used. Multiple correlation coefficients squared (R ²) for minor components stearic acid (0.86), and linolenic acid (0.85) were relatively low because the concentration range is very narrow compared with the reproducibility of the reference method. However, the major FA, oleic (0.99) and linoleic (1.00), FFA (0.97), and chlorophyll (0.98) provided high levels of accuracy. All of the parameters measured were sufficiently accurate for routine screening of olive oil.     

Effects of pH and ferric ions on the antioxidant activity of olive polyphenols in oil-in-water emulsions

The effects of four phenolic compounds occurring in olives and virgin olive oil, namely, oleuropein, hydroxytyrosol, 3,4-dihydroxyphenylethanol-elenolic acid (3,4-DHPEA-EA) and 3,4-dihydroxyphenylethanol-elenolic acid dialdehyde (3,4-DHPEA-EDA), on the oxidative stability of stripped olive oil-in-water emulsions were studied at three pH values in the presence or absence of ferric chloride at 60°C. In the stability test, the addition of phenolic compounds in emulsions at pH 5.5 significantly extended the induction time of lipid oxidation, and the activities in decreasing order were 3,4-DHPEA-EA> 3,4-DHPEA-EDA>hydroxytyrosol>α-tocopherol∼oleuropein ≫control. The effect of concentration, iron, and pH on the antioxidant activity of the phenolic compounds in stripped olive oil-in-water emulsions was analyzed by response surface methodology. Oleuropein and hydroxytyrosol enhanced the prooxidant effect of ferric chloride at pH 3.5 and pH 5.5 but not at pH 7.4. The 3,4-DHPEA-EDA reduced the prooxidant effect of ferric chloride at pH 5.5 and pH 7.4, but at pH 3.5 prooxidant effects were evident at higher phenol concentration. The 3,4-DHPEA-EA reduced the prooxidant effect of ferric ions at all pH values tested. Differences in activity of the phenols may be explained by consideration of their free radical scavenging activity and ferric reducing capacity.     

Chemiotropic behavior of female olive fly (Bactrocera oleae GMEL.) onOlea europaea L

An interpretation is given of a number of observations on the chemiotropic behavior ofBactrocera oleae in connection with olive maceration water and the fly's return to the olive groves after the first summer rains. To this end, the headspace of both maceration water and leaf leaching water, simulating rainfall, were examined. In both cases, the presence of ammonia, which is generally known to attract fruit flies (Diptera, Tephritidae), was detected and, for the first time, in addition to other compounds that are inert for the fly, the presence of styrene was also detected. This aromatic hydrocarbon was found to be a strong attractant. It is shown that both ammonia and styrene are products of the metabolism of microbial flora present on the olive and leaf surface.     

The presence of oxidizing enzyme activities in virgin olive oil

Samples of Greek virgin olive oils were examined for the presence of proteins and oxidative enzyme activities. All oil samples tested contained detectable amounts of protein, as well as lipoxygenase and polyphenol oxidase activities. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and size-exclusion chromatography of olive oil extracts revealed the presence of low-molecular-mass (10–40 kDa) silver-staining and ultraviolet-absorbing components, respectively. Both lipoxygenase and polyphenol oxidase catalytic activities were heat- and protease-sensitive, and they expressed Michaelis-Menten kinetics.     

The triacylglycerol structure of olive oil determined by silver ion high-performance liquid chromatography in combination with stereospecific analysis

The compositions of positionssn-1,sn-2 andsn-3 of triacylglycerols from “extra-virgin” olive oil (Olea europaea) were determined. The procedure involved preparation of diacyl-rac-glycerols by partial hydrolysis with ethyl magnesium bromide; 1,3-, 1,2- and 2,3-diacyl-sn-glycerols as (S)-(+)-1-(1-naphthyl)ethyl urethanes were isolated by highperformance liquid chromatography (HPLC) on silica, and their fatty acid compositions were determined. The same procedure was also carried out on the five main triacylglycerol fractions of olive oil after separation according to the degree of unsaturation by HPLC in the silver ion mode. Although stereospecific analysis of the intact triacyl-sn-glycerols indicated that the compositions of positionssn-1 andsn-3 were similar, the analyses of the molecular species demonstrated marked asymmetry. The data indicate that the “1-random, 2-random, 3-random” distribution theory is not always applicable to vegetable oils.     

Release of ethylene from pruned olive logs: Influence on attack by bark beetles (Coleoptera,Scolytidae)

In recently pruned olive logs, an increase in ethylene release has been observed between 48 and 72 hr after pruning. The values reached, as well as the duration of ethylene release, varied greatly from one log to another. PioneerPhloeotribus scarabaeoides females have shown a preference for logs in which ethylene emission was higher. In logs treated with ethrel, a significant increase in ethylene emission was observed, together with a greater period of release. Therefore, the use of logs treated with ethrel could be of great importance in the control of this pest of olive trees.     

Heat Treatment Improves Olive Oil Extraction

To determine the effect of hot water pre-treatment on olive oil extraction, six cultivars of olive fruit (Olea europaea L. cvs. “Arbequina”, “Hojiblanca”, “Lechín”, “Manzanilla”, “Picual”, and “Verdial”) were heated at 50, 55, and 60 °C prior to laboratory scale oil extraction. Heat treatment resulted in higher oil extraction than unheated control samples. Quality parameters of the oils were not significantly (P < 0.05) affected by these treatments; however, oil stability and bitterness intensity were reduced and pigment content was increased through pre-heat treatment. This process may be incorporated economically into olive oil processing.     

Use of vivianite (Fe3(PO4)2.8H2O) to prevent iron chlorosis in calcareous soils

For various reasons, iron phosphate might be effective in correcting Fe chlorosis in calcareous soils. To test this hypothesis, several pot experiments were conducted using an Fe chlorosis-sensitive chickpea (Cicer arietinum L.) cultivar cropped in soils to which partially oxidized vivianites (Fe₃(PO₄)₂.8H₂O) and Fe(III) phosphates with different characteristics had been added. Vivianites mixed with the soil at a rate of 1 g kg^–1 were as effective in preventing chlorosis as Fe chelate (FeEDDHA). However, the effectiveness of Fe(III) phosphates was less, suggesting that the presence of Fe(II) in the phosphates used was a key factor in their Fe-supplying value to plants. The effectiveness of vivianites, however, seemed to be largely independent of their Fe(II) content.The future of vivianite as a Fe amendment will depend not only on economic considerations (production and application costs) but also on its long-term capacity to release plant-available Fe in soil environments.     

Effect of fruit storage conditions on olive oil quality

"Koroneiki" olive fruit from trees grown in Crete were stored under five different conditions (0°C, air; 5°C, air; 5°C, 2% O₂+5% CO₂; 7.5°C, air; 7.5°C, 2% O₂+5% CO₂). Oil was obtained from fruit immediately after harvest and after fruit storage for 30 and 60 d. Olive oil quality was evaluated by determining acidity, peroxide value, absorption coefficients (K₂₃₂, K₂₇₀), phenol and chlorophyll content, fatty acid composition, and the resistance to oxidation by oven test. Olives stored at 7.5°C, even for 30 d, deteriorated from fungus development, and the obtained oil was of inferior quality with high acidity, peroxide value, and absorption coefficients. The same oil had high chlorophyll and phenol content, resulting in good oil resistance to oxidation. Olive oil from fruit stored at 0 or 5°C for 30 d had acceptable acidity, peroxide value, and absorption coefficients, but showed low resistance to oxidation, which was attributed to low chlorophyll and phenol content. This condition is further attributed to chilling injury caused by low storage temperatures. During storage, all treatments resulted in an increase of oleic acid, partly as a result of linoleic acid oxidation.     

Phenolic composition of virgin olive oils from cross breeding segregating populations

The evaluation and characterization of segregating populations is a critical step in olive breeding programs. In this work, phenolic profiles of virgin olive oils (VOOs) from segregating populations obtained by cross breeding in Cordoba (Spain) have been evaluated. Genotypes obtained from open pollination of the cultivar Manzanilla de Sevilla, and from crosses between the cultivars Arbequina × Arbosana, Picual × Koroneiki and Sikitita × Arbosana were tested. The phenolic composition was determined after liquid–liquid extraction with 60:40 v/v methanol–water and subsequent chromatographic analysis with ultraviolet (UV) detection of both absorption and fluorescence in a sequential configuration. Results for all studied compounds showed high degree of variability between genotypes, with a higher range of variation than the observed for the genitors. Most of the observed variability was attributable to differences in genotypes within crosses rather than among crosses. Some issues related to breeding strategies are discussed. Practical applications: Phenolic compounds are considered to be of paramount importance for the assessment of virgin olive oil quality due to their contribution to the nutraceutical and sensory profile of this natural food. This study focuses on the evaluation of the content of phenolic compounds in olive oils originated from cross breeding in an olive breeding program (Cordoba, Spain). This step is crucial to determine the range of variation of phenolic compounds and the selection of interesting genotypes with higher composition in total phenols or in an individual phenol targeted at a breeding program.     

Variations of iridoid oleuropein in Italian olive varieties during growth and maturation

This research paper reports on the concentrations of oleuropein in fruits of seven Italian olive cultivars. These oleuropein levels, during fruit growth and maturation, were monitored by taking olive samples from the same olive trees grown in Middle Italy. A novel high‐resolution gas chromatography (HRGC) oleuropein quantification method and a new high‐performance liquid chromatography (HPLC) method were developed. The olive fruits showed high contents of oleuropein, which were greatly influenced by the olive cultivar (genetic store) factor. The stage of fruit growth and maturation was another factor strongly affecting the contents of iridoid oleuropein in olive drupes, whatever the olive cultivar considered. Green olives had much higher concentrations of this biofunctional glucoside, which decreased regularly during fruit growth and ripeness. No differences were observed between the two developed oleuropein quantification methods, which gave, for all the olive cultivars, average values of fruit oleuropein contents that were not statistically different. These two reliable repetitive methods, noticeably the HPLC one (which also showed a lower running time), could be used for routine determination of oleuropein in olive drupes. We observed that, in general, olive cultivars having high oleuropein concentrations in the fruits were also characterized by marked oleuropein contents in the leaves.     

Olea europaea Volatiles attractive and repellent to the olive fruit fly (Dacus oleae,Gmelin)

Comparative study of the headspace (HS) composition from olive leaves, olives of varying degrees of ripeness, and crushed olives revealed significant quantitative and qualitative differences. Three different methods were employed to test the effect of the substances identified on the behavior ofDacus oleae gravid females. Attractive/repellent features were tested by presenting the insects with olfactory choices of a water solution of the test chemicals examined, water (blank), and a highly attractive (control) solution of olive maceration water (MW). Toluene and ethylbenzene, present in leaf and half-ripe olive HIS proved attractive, while (E)-2-hexenal emitted by the crushed olives was found to be decidedly repellent. An oviposition stimulation/deterrence test was carried out on olives placed in proximity to test chemicals under examination in water solution, with olives from the same crop serving as control. In the oviposition test-pinene, mostly emitted by the leaves and half-ripe olives, emerged clearly as an activant,p-xylene, myrcenone, ethylbenzene,n-octane ando-xylene as weak activants. (E)-2-hexenal and hexanal, both emitted by the crushed olives, displayed an oviposition deterrent effect. The highly repellent property of (E)-2-hexenal was again confirmed in this test.     

Response of lowland rice and common bean grown in rotation to soil fertility levels on a Varzea soil

Brazil has approximately 30 million hectares of lowland areas, known locally as Varzea, but very little is known about their fertility and crop production potential. A field experiment was conducted for three consecutive years to evaluate response of lowland rice (Oryza sativa L.) grown in rotation with common bean (Phaseolus vulgaris L.) on a Varzea (low, Humic Gley) soil. Rice was grown at low (no fertilizer), medium (100 kg N ha^–1, 44 kg P ha^–1, 50 kg K ha^–1, 40 kg FTE-BR 12 ha^–1), and high (200 kg N ha^–1, 88 kg P ha^–1, 100 kg K ha^–1, 80 kg FTE-BR 12 ha^–1 fritted trace element-Brazil 12 as a source of micronutrients) soil fertility levels. Green manure with medium fertility was also included as an additional treatment. Average dry matter and grain yields of rice and common bean were significantly (P < 0.01) increased with increasing fertilization. Across the three years, rice yield was 4327 kg ha^–1 at low fertility, 5523 kg ha^–1 at medium fertility, 5465 kg ha^–1 at high fertility, and 6332 kg ha^–1 at medium fertility with green manure treatment. Similarly, average common bean yield was 294 kg ha^–1 at low soil fertility, 663 kg ha^–1 at medium soil fertility, 851 kg ha^–1 at high fertility, and 823 kg ha^–1 at medium fertility with green manure treatment. Significant differences in nutrient uptake in bean were observed for fertility, year, and their interactions; however, these factors were invariably nonsignificant in rice.     

Nitrogen fixation by trees in relation to soil nitrogen economy

The N₂-fixing potential (NFP) (i.e. the amount of fixed N₂ in a constraint-free environment) of N₂-fixing trees (NFTs) varies with the genotype. The NFP can be higher than 30-50 g N₂ fixed tree^–1 year^–1 in the most active species, be they leguminous trees such asAlbizia lebbeck, Gliricidia sepium andLeucaena leucocephala, or actinorhizal trees such asCasuarina equisetifolia. The actual amount of nitrogen fixed (ANF) (i.e. the amount of N₂ fixed in the field) is lower than the NFP or even nil because of various constraints, especially drought, nutrient deficiencies, excess of available N and pathogenic nematodes. As tree litters are mineralized, the amount of available N in the soil increases with time, this process leading to the cessation of N₂ fixation in aging plantations. When the mineralization rate is slowed down or inhibited, N₂ fixation can continue. NFTs improve the N status of soils, but the transfer of fixed N to associated plants is not always ensured. Three main approaches are appropriate to increase N₂ fixation: clonal selection of trees combined with vegetative propagation, inoculation with effective rhizobium orFrankia strains, and proper fertilization (especially P). In the absence of major environmental constraints, a positive response to inoculation is expected only when specific (non-promiscuous) NFTs are grown in sites where the density of compatible rhizobia is low or nil. The potentialities of NFTs are far from being fully exploited. Further investigations are proposed and the economics of NFT management is briefly discussed.     

Effect of extraction systems on the quality of virgin olive oil

Research has been carried out to ascertai the effects of different processing systems on olive oil quality. Tests were performed in industrial oil mills that were equipped with both pressure and centrifugation systems. Results show that oils extracted from good-quality olives do not differ in free fatty acids, peroxide value, ultraviolet absorption and organoleptic properties. Polyphenols ando-diphenols contents and induction times are higher in oils obtained from good-quality olives by the pressure system because it does not require addition of water to the olive paste. The centrifugation system requires the addition of warm water to the olive paste and helps to obtain oils with a lower content of natural antioxidants. Oils obtained from poorquality or from ripe olives in continuous centrifugal plants are lower in free fatty acids than those obtained by the pressure system. Dr. Mario Solinas is deceased—May 23, 1993.     

Regional and cultivar comparison of Italian single cultivar olive oils according to flavor profiling

Geographical origin, cultivar, and olive ripening stage are important factors which affect the typical flavor profile of extra virgin olive oils. Aromatic compounds and sensorial profiles of “Casaliva”, “Frantoio”, and “Leccino” olive oils from three different Italian production regions Abruzzo (eastern‐central Italy), Lombardy (northern Italy), and Tuscany (western‐central Italy) were assessed in two cropping years and at three olive ripening stages (green, veraison, and ripe). The chemical aromatic compounds were more effective in discriminating the oils than their sensorial attributes. Oils showed peculiar aromatic and sensorial profiles according to their region of origin. Genetically similar “Casaliva” and “Frantoio” showed also analogous profiles, but distinct from “Leccino”. The interaction between regions of cultivation and cultivars significantly affected the flavor profiling. Practical applications: This research provides a sound proof that geographical origin, cultivar, and fruit ripening stage, could play an important interactive role in shaping the flavor profiling of extra virgin olive oils. This represents a scientific basis of the “typicality” concept, which should address the marketing strategies for exploiting of the PDO products.