Effect of different olive oil-derived antioxidants (hydroxytyrosol and 3,4-dihydroxyphenylglycol) on the quality of frozen-thawed ram sperm

The aim of the present study was to evaluate the effect of the addition of different concentrations of two olive oil-derived antioxidants, hydroxytyrosol (3,4-dihydroxyphenylethanol, HT) and 3,4-dihydroxyphenylglycol (DHPG), on ovine semen during the freezing-thawing process. Sperm was collected, pooled and diluted with commercial extenders and then divided into aliquots supplemented with different concentrations (10 μg/ml, 30 μg/ml, 50 μg/ml and 70 μg/ml) of HT, DHPG and a mixture (MIX) of both antioxidants. A control group, without antioxidant, was also prepared. Sperm motility, viability, acrosome integrity, mitochondrial membrane potential and lipid peroxidation (LPO) were assessed. The results showed that frozen-thawed ram spermatozoa exhibited lower values for motility, membrane integrity, acrosome and mitochondrial membrane potential than fresh samples (P ≤ 0.01). However, when antioxidants were added, thawed spermatozoa exhibited relatively low LPO, recording values similar to fresh spermatozoa; by contrast, the control group of frozen-thawed spermatozoa without antioxidants exhibited significantly higher LPO (P ≤ 0.01). The addition of a HT+DHPG mixture (MIX) had a negative impact on sperm membrane and acrosome integrity, suggesting that a pure antioxidant supplementation has the potential to offer superior results. In conclusion, HT and DHPG exhibited a positive effect on the frozen-thawed spermatozoa inasmuch as they reduced the LPO. These olive oil-derived antioxidants have the potential to improve frozen-thawed sperm quality, although further studies should be carried out to analyse the antioxidant effect at different times after thawing.     

The combined effects of black oxidising table olive process and ripening on the cell wall polysaccharides of olive pulp

Olive fruits harvested at cherry and black stages of ripening were processed according the table olive black oxidising processing and sampled after the three main steps: storage in brine, lye treatment and thermal treatment (final product). The results show that the storage in brine contributed positively to the stabilisation of cell wall polysaccharides of olive pulp as the amounts of main polysaccharides practically were maintained in both stages of ripening. The lye treatment introduced degradation of cell walls due to the generalised loss of pectic and hemicellulosic polysaccharides and cellulose, caused by the breakage of ester and hydrogen bonds. On the other hand, the lye treatment introduced shifts in the solubilisation of polysaccharides rendering them more difficult to extract by alkali solutions and enabling their retention in the cellulosic residue, which should contribute positively to cell wall firmness. The thermal treatment introduced degradation of cellulose and increased the solubilisation of polysaccharides with a higher extent in the black olives. This work showed that the differences on the cell wall polysaccharides between stages of ripening are magnified after processing and allowed to conclude that the stage of ripening of olive fruits is determinant for obtaining a final product with adequate texture properties for table olive consumption.     

Characterization of olive seed storage proteins

At present little is known about olive seed storage proteins (SSPs). A better understanding of olive SSPs will be important for future biotechnology efforts. In the present study, we first developed a protocol relied on chloroform for preparing protein samples free of lipids from lipid-rich olive seeds. Then, we characterized olive SSPs by SDS-PAGE, N-terminal sequencing and immunoblot. Two smaller subunits (20 and 21.5 kD) of SSPs were purified to homogeneity and used for antibody production or N-terminal sequencing. N-terminal sequencing confirmed that major olive SSPs are 11S globulins. Moreover, the components and size distribution of SSPs are identical among several olive cultivars examined, suggesting that their synthesis is highly conserved in this species. Olive SSPs are soluble in aqueous alcohol, with limited solubility in water and dilute salt. Thus, despite their homology with globulins, olive SSPs are similar in solubility to prolamins and different from globulins in other dicot plants. Finally, the accumulation of olive SSPs during fruit maturation was examined. Our results revealed that the accumulation of SSPs is time-dependent and tissue-specific, and only 105 days after pollination (DAP), did individual components of SSPs synthesize substantially, and accumulate rapidly in large quantities over a short period of time. Our results suggest that a 36 kD protein is the precursor of olive SSPs, and 90–105 DAP seems to be a crucial transition period (from a precursor to mature subunits) for the accumulation of SSPs.     

Assessment of the genotoxicity of olive mill waste water (OMWW) with the Vicia faba micronucleus test

The present study concerns the genotoxicity of olive mill waste water (OMWW) generated in mills producing olive oil in Morocco. The Vicia faba micronucleus test was used to evaluate the genotoxicity of OMWW and the six major phenolic compounds identified by HPLC in this effluent. Five dilutions of OMWW were tested: 0.1, 1, 5, 10 and 20%. Maleic hydrazide was used as a positive control. The results showed that OMWW was genotoxic at 10% dilution. In order to investigate the components involved in this genotoxicity, the six major phenols present in this effluent, oleuropein, gallic acid, 4-hydroxyphenyl acetic acid, caffeic acid, paracoumaric acid and veratric acid, were studied at concentrations corresponding to the genotoxic concentration of the OMWW itself. Two phenols, gallic acid and oleuropein induced a significant increase in micronucleus frequency in Vicia faba; the four other phenols had no significant genotoxic effect. These results suggest that under the experimental conditions of our assay, OMWW genotoxicity was associated with gallic acid and oleuropein.     

Chloroplast-DNA variation in cultivated and wild olive (Olea europaea L.)

Polymorphism in the lengths of restriction fragments of the whole cpDNA molecule was studied in cultivated olive and in oleaster (wild olive) over the whole Mediterranean Basin. Seventy two olive cultivars, 89 very old trees cultivated locally, and 101 oleasters were scored for ten endonucleases. Moreover, maternal inheritance of cpDNA in olive was shown by analysing the progeny of a controlled cross between two parents which differed in their cpDNA haplotypes. In the whole species, three site- and three length-mutations were observed, corresponding to five distinct chlorotypes. The same chlorotype (I) was predominant in both oleasters and cultivated olive trees, confirming that these are closely related maternally. Three other chlorotypes (II, III and IV) were observed exclusively in oleaster material and were restricted either to isolated forest populations or to a few individuals growing in mixture with olive trees possessing the majority chlorotype. An additional chlorotype (V) was characterised by three mutations located in distinct parts the cpDNA molecule but which were never observed to occur separately. This chlorotype, more widely distributed than the other three, in both cultivated and wild olive, and occurring even in distant populations, was observed exclusively in male-sterile trees showing the same specific pollen anomaly. However, in the present study, no evidence was provided for a direct relationship between the occurrence of the cpDNA mutations and male sterility. It is suggested that the large geographic distribution of chlorotype V may be related to the high fruit production usually observed on male-sterile trees. These may be very attractive for birds which are fond of olive fruit and spread the stones efficiently. Probably for the same reason, people preserved male-sterile oleasters for long periods and, in several places, used male-sterile cultivars over large areas. Received: 25 November 1998 / Accepted: 19 December 1998     

Changes in water relations,photosynthetic activity and proline accumulation in one-year-old olive trees (<Emphasis Type="Italic">Olea europaea</Emphasis> L. cv. Chemlali) in response to NaCl salinity

The comparative responses of young olive trees (Olea europaea L. cv “Chemlali”) to different NaCl salinity levels were investigated over 11 months. One-year-old own rooted plants were grown in 10-L pots containing sand and perlite mixture (1:3 v/v). Trees were subjected to three irrigation treatments: CP (control plants that were irrigated with fresh water); SS1 (salt stressed plants irrigated with water containing 100 mM NaCl) and SS2 plants (salt stressed plants irrigated with water containing 200 mM NaCl). Shoot elongation rate, relative water content, leaf water potential and net carbon dioxide exchange rates decreased significantly with increased NaCl salinity level. Under stressed conditions, the increase of Na⁺ and Cl⁻ ions in both leaves and roots was accompanied with that of proline and soluble sugars. The above results show that the accumulation of proline and sugars under stressed conditions could play a role in salt tolerance. The absence of toxicity symptoms under both stress treatments and the superior photosynthetic activity recorded in SS1-treated plants suggest that cv Chemlali is better able to acclimatize to 100 mM NaCl than at 200 mM NaCl. Our findings indicate that saline water containing 100 mM NaCl, the most available water in arid region in Tunisia, can be recommended for the irrigation of cv Chemlali in the arid south of Tunisia.     

The fate of olive oil polyphenols in the gastrointestinal tract: Implications of gastric and colonic microflora-dependent biotransformation

We have conducted a detailed investigation into the absorption, metabolism and microflora-dependent transformation of hydroxytyrosol (HT), tyrosol (TYR) and their conjugated forms, such as oleuropein (OL). Conjugated forms underwent rapid hydrolysis under gastric conditions, resulting in significant increases in the amount of free HT and TYR entering the small intestine. Both HT and TYR transferred across human Caco-2 cell monolayers and rat segments of jejunum and ileum and were subject to classic phase I/II biotransformation. The major metabolites identified were an O-methylated derivative of HT, glucuronides of HT and TYR and a novel glutathionylated conjugate of HT. In contrast, there was no absorption of OL in either model. However, OL was rapidly degraded by the colonic microflora resulting in the formation of HT. Our study provides additional information regarding the breakdown of complex olive oil polyphenols in the GI tract, in particular the stomach and the large intestine.     

Quantitative forecasting of olive yield in Northern Portugal using a bioclimatic model

In this work the objective was to develop a bioclimatic model to forecast olive yield based on airborne pollen, soil water content, and favourable conditions for phytopathological attacks. Olive airborne pollen was sampled from 1998 to 2006 using Cour traps installed in the Trás-os-Montes e Alto Douro region, in the provinces of Valença do Douro and Vila Nova de Foz-Côa. Meteorological data from a meteorological station located in Pinhão, near the pollen samplers, was used to calculate other independent variables. According to the bioclimatic model, at the flowering stage 63% of regional olive production can be predicted from the regional pollen index, with an average deviation between observed and predicted production of 10%. The variable soil water content enabled an increase in forecasting accuracy of about 30%, and a reduction in the average deviation between observed and predicted production of 6%. The final regression with all three variables tested showed that the bioclimatic model was able to predict the annual variability of regional olive fruit production with an accuracy of 97%, the average deviation between observed and predicted production being 3% for internal validation and 6% for external validation.     

Physiological effects of the induction of resistance by compost or Trichoderma asperellum strain T34 against Botrytis cinerea in tomato

Certain types of compost used as growth media can induce resistance to foliar pathogens in above-ground parts of a plant. The induction of resistance can sometimes be associated with growth impairment and yield reduction. The objective of this study was to establish whether plants grown in olive marc compost had enhanced resistance against Botrytis cinerea at the cost of growth or physiological performance.Tomato plants grown in mature olive marc compost had approximately 60% less disease severity than plants grown in perlite. As a reference, plants grown in perlite enriched with the known inducer of resistance Trichoderma asperellum strain T34 (T34) had 35% less disease severity than plants grown in perlite. The salicylic acid (SA) pathway/abscisic acid (ABA) is involved in compost induced systemic resistance. Instead, perlite enriched with T34 is not linked to SA pathway/ABA. Physiological measures of water status, root/shoot ratio, stable isotopes of C and chlorophyll fluorescence showed that the plants grown in compost were close to a stress situation. However, growth measured as biomass and plant height of plants grown in compost was higher than in plants grown in perlite suggesting that plants in compost were not grown in a stress situation, but in a eustress. Tomato plants grown in perlite enriched with T34 had better growth, measured as total leaf area, biomass, height and nutrient uptake, than plants grown in perlite. Physiological measures showed that plants grown either in perlite or perlite enriched with T34 did not show any abiotic stress situation.     

Oleo‐Signs and Quali‐Signs: The Qualities of Olive Oil

This article considers the way that a single substance, olive oil, bundles together a set ofqualisigns including luminosity, liquidity, spreadibility, durability, capacity to cleanse, capacity to seal or preserve, capacity to insulate, and notably, a lack of miscibility in water. These qualisigns allow olive oil to operate in several discrete religious contexts in the Mediterranean although the interpretations and the specific qualisigns made salient by particular semiotic metadiscourses are always changing, even though the material substrate remains the same. In contemporary Western discourses of distinction, the positive qualities of olive oil are not those primarily registered by the ordinary senses of taste, touch, and smell, but rather those that are established discursively by expert opinions of various kinds, which attest to olive oil's ‘healthful’ qualities.