Antioxidant Effect of Natural Table Olives Phenolic Extract Against Oxidative Stress and Membrane Damage in Enterocyte-Like Cells.

The phenolic fraction of a naturally fermented cultivar of table olives, "Tonda di Cagliari," was investigated for the ability to protect Caco-2 cells against oxidative stress and membrane damage induced by tert-butyl hydroperoxyde (TBH). TBH exposure resulted in an alteration of cellular redox status, with an increase in reactive oxygen species (ROS) and a decrease in reduced glutathione (GSH) level. A loss of the epithelial integrity, as indicated by the decrease of the transepithelial electrical resistance value, was also observed over time, together with an intense lipid peroxidation process. The olives phenolic extract significantly counteracted ROS generation and subsequent alteration of monolayer integrity and membrane oxidative damage. The protective action of the extract is likely due to the scavenging ability of its main components, as hydroxytyrosol, oleuropein, and verbascoside among the secoiridoids and derivatives. Since olives phenolic compounds concentrate in the intestinal lumen, they may be a useful tool in the prevention of intestinal disorders related to oxidative damage.

Nematicidal activity of 2-thiophenecarboxaldehyde and methylisothiocyanate from caper (Capparis spinosa) against Meloidogyne incognita.

New pesticides based on plant extracts have recently gained interest in the development of nontoxic crop protection chemicals. Numerous research studies are focused on the isolation and identification of new active compounds derived from plants. In this manuscript we report about the use of the Mediterranean species Capparis spinosa as a potent natural nematicidal agent against the root knot nematodes Meloidogyne incognita. Leaves, stems, and caper buds of Capparis spinosa were used to obtain their methanol extracts (LME, SME, BME) that were successively in vitro tested against second stage nematode juveniles (J2). In terms of paralysis induction, the methanol extract of the stem part (SME) was found more effective against M. incognita and then the caper methanol buds and leaves extracts. The chemical composition analysis of the extracts carried out by GC/MS and LC/MS techniques showed that methylisothiocyanate was the main compound of SME. The EC50 for SME after 3 days of immersion was 215 ± 36 mg/L. The constituent components of SME such as 2-thiophenecarboxaldehyde and methylisothiocyanate were successively in vitro tested for their nematicidal activity against J2. Both compounds induced paralysis on root knot nematodes ranking first (EC50 = 7.9 ± 1.6, and 14.1 ± 1.9 mg/L respectively) for M. incognita. Moreover, 2-thiophenecarboxaldehyde showed a strong fumigant activity.

Nematicidal activity of (E,E)-2,4-decadienal and (E)-2-decenal from Ailanthus altissima against Meloidogyne javanica.

Methanol extracts of various plant parts of Ailanthus altissima were tested against the root knot nematode Meloidogyne javanica . Extracts of bark (ABE), wood (AWE), roots (ARE), and leaves (ALE) from A. altissima were investigated against freshly hatched second-stage juveniles (J(2)). AWE was the most active extract, with EC(50/3d) of 58.9 mg/L, while ALE, ARE, and ABE did not show nematicidal activity. The chemical composition of the extracts of A. altissima was determined by gas chromatography-mass spectrometry, and (E,E)-2,4-decadienal, (E)-2-undecenal, (E)-2-decenal, hexanal, nonanal, and furfural were the most prominent constituents. (E,E)-2,4-Decadienal, (E)-2-decenal, and furfural showed the highest nematicidal activity against M. javanica , with EC(50/1d) = 11.7, 20.43, and 21.79 mg/L, respectively, while the other compounds were inactive at the concentrations tested. The results obtained showed that AWE and its constituents (E,E)-2,4-decadienal and (E)-2-decenal could be considered as potent botanical nematicidal agents.

LC/DAD/ESI/MS method for the determination of imidacloprid, thiacloprid, and spinosad in olives and olive oil after field treatment.

The behavior in the field and the transfer from olives to olive oil during the technological process of imidacloprid, thiacloprid, and spinosad were studied. The extraction method used was effective in extracting the analytes of interest, and no interfering peaks were detected in the chromatogram. The residue levels found in olives after treatment were 0.14, 0.04, and 0.30 mg/kg for imidacloprid, thiacloprid, and spinosad, respectively, far below the maximum residue levels (MRLs) set for these insecticides in EU. At the preharvest interval (PHI), no residue was detected for imidacloprid and thiacloprid, while spinosad showed a residue level of 0.04 mg/kg. The study of the effect of the technological process on pesticide transfer in olive oil showed that these insecticides tend to remain in the olive cake. The LC/DAD/ESI/MS method showed good performance with adequate recoveries ranging from 80 to 119% and good method limits of quantitation (LOQs) and of determination (LODs). No matrix effect was detected.

Control of postharvest diseases of fruit by heat and fungicides: efficacy, residue levels, and residue persistence. A review.

Extensive research has been done in recent years to reduce the heavy dependence on chemical fungicides to control postharvest diseases and disorders of horticultural crops. Alternative strategies were based on improved cultural practices, biological control, plant-defense promoters, and physical treatments such as UV illumination, radiofrequency treatment, heat therapy, and storage technologies. Among these, postharvest heat treatments such as hot water dips, short hot water rinsing and brushing, and hot air conditioning have reduced rot development and enhanced fruit resistance to chilling injury in sensitive cultivars while retaining fruit quality during cold storage and shelf life. Additive or synergistic increases in effectiveness were observed by integrating heat therapy with various chemical compounds, thus leading to significant reductions in the application of active ingredients to protect produce from decay. This paper highlights the knowledge on this topic with emphasis on heat therapy effects and factors affecting the uptake, persistence, and performance of fungicide residues when they are applied in combination with hot water.

Chlorpyrifos residues levels in fruits and vegetables after field treatment.

Chlorpyrifos (O,O-diethyl O-3,5,6-trichloro-2-pyridyl phosphorothioate) was applied with three different formulations on oranges, peaches, tomatoes, wine and table grapes, and its behaviour was evaluated after field treatment. The formulations applied were emulsifiable concentrates (EC), microencapsulates (ME), and wettable granules (WG). The residues were similar in all crops studied in the EC and WG experiments, except peaches with WG treatment, the residue amount was lower than EC values. Tomatoes which were grown in greenhouse showed similar residues in all treatments just after treatment. Wine and table grapes showed different decline curves in the EC experiments ascribable to the different growing technology. Instrumental limit of determination (LOD) and limit of quantification (LOQ) for all matrices were 0.01, and 0.03 mg kg⁻¹, respectively. Repeated treatments showed that Chlorpyrifos can accumulate leading to residue levels at the preharvest interval (PHI) over the maximum residue level (MRL), especially on oranges and peaches. Among the formulates used ME showed the higher risk of residues over the MRL at harvest.

Chemical variability, antifungal and antioxidant activity of Eucalyptus camaldulensis essential oil from Sardinia.

Essential oil (EO) from aerial parts of Eucalyptus camaldulensis Dehnh., growing wild in different localities of Sardinia (Italy), was extracted by steam distillation and analyzed by gas chromatography (GC) FID and GC-ion trap mass spectrometry (ITMS). The yields of EO (v/dry wt) ranged between 0.2-0.5%. Samples were harvested between April and December to study the seasonal chemical variability of the EO. The chemical composition varied depending on the different origins and showed strong fluctuation during the vegetative stage. Thirty-seven compounds, accounting for at least 97.7% of the total EOs were identified, the major components being p-cymene (27.8-42.7%), 1,8-cineole (4.1-39.5%), beta-phellandrene (3.9-23.8%), spathulenol (2.1-15.5%) and cryptone (3.2-10.2%). The oils possessed moderate amounts (1.4-4.7%) of two uncommon aldehydes, cuminal and phellandral. The essential oils were screened for their antifungal activities against common phytopathogenic fungi using the paper disk diffusion method and they showed activity at low doses against the fungi tested. The antioxidant activity, assessed by DPPH-test and expressed as Trolox equivalent antioxidant capacity, showed values ranging between 0.5 and 5.8 mmol/L.

Influence of post-harvest application rates of cyprodinil, treatment time and temperature on residue levels and efficacy in controlling green mould on 'Valencia' oranges.

The effectiveness of heat treatments with water and cyprodinil in controlling post-harvest green mould caused by Penicillium digitatum was investigated on artificially inoculated 'Valencia' oranges. Residue levels of cyprodinil were determined in the oranges as a function of active ingredient concentration, temperature and treatment time. Cyprodinil residues were significantly dependent on treatment time when applied at 600 mg l(-1) and 20 degrees C, but not when fruit were treated at 150-300 mg l(-1). The application of cyprodinil at 50 or 100 mg l(-1) at 55 degrees C for 30 s produced similar residue levels, while residues increased when the application rate was 150 mg l(-1). Cyprodinil at 100 mg l(-1) and 60 degrees C produced a significant increase in residues compared to treatment at 50 mg l(-1); no significant increase in residues was found when the application rate was raised from 100 to 150 mg l(-1). In comparison to treatments performed at 20 degrees C, the application of a heated cyprodinil mixture resulted in significantly higher residues in fruit. All treatments with cyprodinil at 20 degrees C similarly reduced green mould after 7 days of storage at 20 degrees C. After 18 days, treatment with cyprodinil at 600 mg l(-1) for 30 s was more effective than at 150-300 mg l(-1). When dip time was extended to 90 or 180 s, treatment efficacy was positively related to fungicide concentration. Treatments with water at 55 degrees C for 30 s were as effective as cyprodinil at 50-100 mg l(-1), but less effective than cyprodinil at 150 mg l(-1). After 7 days, treatment with water or cyprodinil at 50-150 mg l(-1) and 60 degrees C were equally effective in controlling green mould; while, after 18 days, treatment with cyprodinil at 150 mg l(-1) was consistently more effective than at 50-100 mg l(-1) or hot water alone.

Fate of azadirachtin A and related azadirachtoids on tomatoes after greenhouse treatment.

The degradation of the main azadirachtoids on tomatoes was studied after greenhouse treatment. These experiments were carried out at 1 and 5x the concentration recommended by the manufacturer. In all experiments the deposition of azadirachtin A (AZA-A) was below the maximum residue level (MRL). Even if at the highest dose, AZA-A half-life time calculated as pseudo first order kinetic was 1.2 days in agreement with the recommended preharvest interval (PHI) of 3 days. Experiments with a model system showed that sunlight photodegradation is the main factor influencing the rate of disappearance of AZA-A after greenhouse treatment while tomato epicuticular waxes doubled the photodegradation rate of AZA-A in a commercial formulation.

Influence of postharvest hot water treatment on nutritional and functional properties of kumquat (Fortunella japonica Lour. Swingle Cv. Ovale) fruit.

The present study investigated the influence of a hot water dip (HWD) for 2 min at 50 degrees C, a standard and effective treatment for postharvest decay control of citrus fruit, on the nutritional and health-related properties of kumquats. The results show that most of the parameters examined, including titratable acidity, soluble solids content, maturity index, glucose, fructose, sucrose, ascorbic acid, dehydroascorbic acid, alpha- and gamma-tocopherols, beta-carotene, zeaxantin, rhoifolin, and antioxidant activity, were not significantly affected by treatment. The levels of beta-cryptoxanthin, narirutin, and total flavonoids increased after HWD, whereas lutein and total phenols decreased. The concentration of the essential oil and the relative percentage of the individual components of the essential oil were not affected by HWD except for the minor compound p-menta-1,5-dien-1-ol, which increased after HWD. After storage, lower levels of glucose, total sugars, beta-carotene, beta-cryptoxanthin and lutein were recorded in HWD fruit. A decrease in antioxidant activity and increases in alpha-tocopherol and total vitamin E were found both in control and HWD fruit. The influence of HWD at 50 degrees C for 2 min on individual nutraceuticals and health-related properties was thus generally low and may depend on storage conditions.

Antimicrobial activity of Tunisian quince (Cydonia oblonga Miller) pulp and peel polyphenolic extracts.

Quince (Cydonia oblonga Miller) fruit aqueous acetone extracts were evaluated. High-performance liquid chromatography-diode array detection and electrospray ionization-mass spectrometry were used for the identification and quantification of the phenolic compounds. The total phenolic content of the pulp and peel parts ranged from 37 to 47 and 105 to 157 mg/100 g of fresh weight, respectively. Chlorogenic acid (5-O-caffeoylquinic acid) was the most abundant phenolic compound in the pulp (37%), whereas rutin (quercetin 3-O-rutinoside) was the main one in the peel (36%). The radical scavenging potential of the extracts was determined and compared with that of synthetic antioxidants. The stronger properties corresponded to those obtained from peel material with a 70-80% inhibitory effect on DPPH radicals. The antimicrobial activity of the extracts against different microorganism strains was also investigated. Quince peel extract was the most active for inhibiting bacteria growth with minimum inhibitory and bactericide concentrations in the range of 102-5 x 103 microg polyphenol/mL. It seems that chlorogenic acid acts in synergism with other components of the extracts to exhibit their total antimicrobial activities.

Residues and persistence of neem formulations on strawberry after field treatment.

Azadirachtoids were determined by liquid chromatography/mass spectrometry (LC/MS) in five methanolic seed extracts of the neem tree and in a commercial formulation. On average, seed extracts contain azadirachtin A (10.9%), azadirachtin B (3.5%), nimbin (10.4%), and large quantities of salannin (19.0%). The composition of the commercial formulations may present different azadirachtoids contents depending on the natural extracts used in the preparation. Because these compounds may also show insecticide activity, the efficacy on field of these formulations may be very different. Photodegradation of pure azadirachtoids was also studied. Azadirachtins and related compounds are very sensitive to sunlight, degrading rapidly, with half-lives of the order of 11.3 h for azadirachtin A and 5.5 h for azadirachtin B and few minutes for the other limonoids compounds studied. The residues of azadirachtins and the main constituents, e.g., salannin, nimbin, deacetylnimbin, and deacetylsalannin, of the neem seed extract were determined on strawberries after field treatment using two different formulations. This residue study on strawberry was carried out to assess not only the azadirachtin content but also the main azadirachtoids contents. Three days after field application at five times the dose recommended by the manufacturer, residues of azadirachtin A and B were 0.03 and 0.01 mg/kg, respectively, while residues of salannin (LOQ 0.01 mg/kg) and nimbin (LOQ 0.5 mg/kg) were not detectable.

Chemical composition, seasonal variability, and antifungal activity of Lavandula stoechas L. ssp. stoechas essential oils from stem/leaves and flowers.

Essential oils from the stems/leaves (L) and flowers (F) of Lavandula stoechas L. ssp. stoechas growing wild in southern Sardinia (Italy) were extracted by hydrodistillation and analyzed by gas chromatography coupled with flame ionization detector and ion trap mass spectrometry. The major compound was fenchone, accounting for, on average, 52.60% in L and 66.20% in F, followed by camphor (13.13% versus 27.08%, in L and F, respectively). F essential oil yields (volume per dry weight) decreased from the beginning to the end of the flowering stage, whereas L yields remained constant during the year. The nine main compounds derived from two different subpathways, A and B. The compounds that belong to the same subpathway showed a similar behavior during the year. The essential oils were tested for their antifungal activity using the paper disk diffusion method. The essential oils tested were effective on the inactivation of Rhizoctonia solani and Fusarium oxysporum and less effective against Aspergillus flavus. Among the single compounds tested, fenchone, limonene, and myrtenal appeared to be the more effective on the inhibition of R. solani growth.

Chemical composition of volatiles in Sardinian myrtle (Myrtus communis L.) alcoholic extracts and essential oils.

The chemical composition of the volatile fraction of myrtle (Myrtus communis L.) alcoholic extracts and essential oils from leaves and berries collected in different places in Sardinia (Italy) was studied. A simple and rapid liquid-liquid extraction method was used to isolate volatile compounds from myrtle alcoholic extracts followed by GC and GC-MS analysis allowing the detection of 24 compounds. The volatile fraction was characterized by the terpenes fraction corresponding to that of the essential oils and by a fatty acid ethyl esters fraction. The variation during extraction of the volatile fraction in alcoholic extracts of berries and leaves was evaluated. Essential oils were obtained by hydrodistillation, and the yields were on average 0.52 +/- 0.03% (v/w dried weight) and 0.02 +/- 0.00% for leaves and berries, respectively. The essential oils were analyzed by GC and GC-MS, and a total of 27 components were detected, accounting for 90.6-98.7% of the total essential oil composition. Strong chemical variability depending on the origin of the samples was observed. The major compounds in the essential oils were alpha-pinene (30.0 and 28.5%), 1,8-cineole (28.8 and 15.3%), and limonene (17.5 and 24.1%) in leaves and berries, respectively, and were characterized by the lack of myrtenyl acetate.

Chemical composition, plant genetic differences, and antifungal activity of the essential oil of Helichrysum italicum G. Don ssp. microphyllum (Willd) Nym.

The chemical composition of the essential oil of the Sardinian dwarf curry plant [Helichrysum italicum G. Don ssp. microphyllum (Willd) Nym] was studied. Genetic analysis suggested the presence of two chemotypes; morphological and chemical differences confirmed the presence of two chemotypes (A and B). The maximum yields were 0.18 and 0.04% (v/w) for flowering tops and stems, respectively. The concentrations of nerol and its esters (acetate and propionate), limonene, and linalool reach their highest values during the flowering stage both in flowers and in stems. Besides the essential oil, type B showed an interesting antifungal activity.

Persistence of azadirachtin residues on olives after field treatment.

In this work an HPLC method for the determination of azadirachtin residues on olives was developed, and the field degradation kinetics of the pesticide was studied. In field trials the active ingredient (a.i.) decay had a half-life time of 0.8 days, which was too short to show a good efficacy of treatment. The mechanism of disappearance of the pesticide studied with model systems showed that it was unrelated to evaporation, thermodegradation, and co-distillation, but it was related to photodegradation. The high photodegradation rate of commercial formulations calls for the need to test different formulates in order to increase the persistence of the residue and thus the pesticide's efficacy.

Residue uptake and storage responses of Tarocco blood oranges after preharvest thiabendazole spray and postharvest heat treatment.

Tarocco blood oranges (Citrus sinensis Linn. Obsek) were subjected to a single preharvest spray with thiabendazole (TBZ) at a concentration of 1% active ingredient (ai) in water and harvested 2 weeks after fungicide application or heated at 37 degrees C for 48 h under saturated humidity after harvest. The two treatments were also combined before cold quarantine (3 weeks at 2 degrees C), subsequent storage (3 weeks at 8 degrees C), and simulated marketing period (SMP) (1 week at 20 degrees C). Fruit not treated with TBZ and unheated were used as controls. The residue levels of TBZ (active ingredient, whole fruit basis) after spray were approximately 6.3 and 5.4 mg x kg(-1) before fruit storage respectively, a level close to the tolerance limit set by the European Community. TBZ showed a high persistence during quarantine, storage, and SMP. TBZ spray significantly reduced the incidence and severity of chilling injury (CI) and decay during the postquarantine period and SMP. Heat treatment (HT) produced beneficial effects in controlling CI, especially during SMP, when applied in combination with TBZ. However, HT remarkably promoted the development of secondary fungal infections such as Phytophthora rots and adversely affected fruit flavor and taste. The occurrence of off-flavor and off-taste was found to be perceptible after heating.

Rotenone residues on olives and in olive oil.

The disappearance of rotenone on olives under field conditions was studied. The field data showed that rotenone residues on olives decreased with a half-life (t(1/2)) of 4.0 days. After pre-harvest time (10 days) the residues were higher than the maximum residue level fixed in Italy (0.04 mg/kg). Experiments with model systems showed that the mechanism of disappearance of rotenone is not related to evaporation, thermodegradation, or co-distillation, but only to photodegradation. When the olives were processed for oil, the residues in the oil were higher than the residues on the olives by a factor of 2.4-4.8.