Model study on the effect of 15 phenolic olive mill wastewater constituents on seed germination and Vibrio fischeri metabolism

Olive mill wastewaters (OMW) can be a severe problem when disposed of as untreated because of their high organic load, elevated concentration of polyphenols, and moderately low biodegradability. In the present study, the acute toxicity of 15 compounds with low molecular weight (<350 Da), catechol, four benzoic acids, three phenylacetic acids, three phenylethanols, and four cinnamic acids, already isolated from the reverse osmosis in the fractionation of OMW, was assessed on the marine bacterium Vibrio fischeri and on the seeds of two dicotyledonous species Cucumis sativus and Lepidium sativum, and on one monocotyledon Sorghum bicolor. Results of phytotoxicity showed that the most toxic compounds were catechol (EC50s ranging from 0.40 mmol/L for S. bicolor to 1.09 for C. sativus) and hydroxytyrosol, (EC50s ranging from 0.47 mmol/L for S. bicolor to 1.55 for C. sativus) while the toxic potential on bacteria was particularly elevated with EC50 values 1 or 2 orders less than phytotoxicity. These results suggested that the risk of OMW disposal may be more elevated for the water compartment than for the soil.     

Yield and Quality of Leek in Response to Compost Applied as a Mulch or Incorporated Into the Soil

Compost is widely used to increase soil fertility, usually practiced by incorporating the compost into the upper soil layer. This study questions the rationale behind this practice. Compost was applied as a mulch and compared with compost worked into the soil in a growth experiment with leek (Allium porrum L. Var. ‘Siegfried Frost’). The compost used was made of source separated organic waste from either gardens and parks, or households. Garden-park compost was applied in 2.5 times greater volumes than household compost to compensate for its lower content of nutrients. The soil was either sandy loam or clay loam. Each of the eight combinations of variables (application method, compost type, and soil type) was repeated three times with 20 leeks in each replicate. Significantly higher yields were obtained with compost applied as a mulch. Here, the yield averaged 78 g fresh weight per leek, compared to 59 g per leek from plots with compost incorporated. Compost mulching also resulted in a significantly higher quality leeks, including more first class leeks, longer and thicker shafts, and a generally better appearance. The advantage of placing the compost on the soil surface rather than thoroughly mixing it with the soil can be attributed to a higher availability of plant nutrients. No significant effect of compost type on leek yield was observed, indicating that the 2.5 times higher volumetric dose of garden-park compost provided the same amount of available nutrients as a single dose of household compost. The soil type did not significantly influence the yields either, which is attributable to both soils being well structured prior to compost amendment.     

Exotic vs. native plant dominance over 20 years of old-field succession on set-aside farmland in Argentina

Exotic plants are major constituents of species pools in modern landscapes. Managing succession for restoration of degraded ecosystems thus requires an understanding of novel trajectories unfolding in mixed, native/exotic plant assemblages. We examined trends in native and exotic species abundance over 20 years of old-field succession on set-aside farmland in the Inland Pampa, Argentina. Changes in plant cover and species richness were annually monitored on adjacent permanent plots established in different years (1978-1989). Both native and exotic species occurred in early, mid and late successional stages, exhibiting similar life-form replacement patterns, from annual forbs, through annual to perennial grasses. Exotic plant richness declined with plot age. Yet, four exotic grasses remained dominant through succession (50-70% cover), with plots initiated in later years showing increased exotic cover. While native perennial grasses occurred from the onset of succession, increasing from 5 to 12 spp/plot, they only showed transient peaks below 30% cover. Cluster analysis of 113 plot-year samples identified alternative community states for early, mid and late successional stages, which were connected by a complex network of interweaving dynamic pathways. Depending on the plot, vegetation dynamics comprised directional temporal trajectories as well as nondirectional pathways, and arrested community states dominated by exotic grasses. Our results illustrate the overwhelming role of exotic species in modern old-field succession, and their potential to hinder recovery of native communities on former agricultural land. Community states with novel, native/exotic plant mixtures could be managed to deliver specific ecosystem services (e.g. forage production, carbon sequestration). However, meeting conservation goals may require active restoration measures, including exotic plant removals and native grass seeding.     

Evaluation of leaf rust resistance genes Lr1 , Lr9 , Lr24 , Lr47 and their introgression into common wheat cultivars by marker-assisted selection

Epidemiological field controls in different Italian locations and seedling evaluations of the ‘Thatcher’ near-isogenic lines (NILs) carrying the leaf rust resistance genes Lr1, Lr9, Lr24 and Lr47 were conducted during 5 years of testing. These genes confirmed their effectiveness in both field and greenhouse conditions. Moreover a backcross program was carried out by using as recurrent parents the susceptible high-quality common wheat cvs ‘Bolero’, ‘Colfiorito’, ‘Serio’ and ‘Spada’ and the ‘Thatcher’ NILs carrying the above mentioned genes as donor parents. The progenies of different cross combinations were selected by both resistance tests and marker assisted selection using molecular markers (STS, SCAR, CAPS) closely linked to Lr genes: a complete cosegregation was observed between the resistance genes used and the corresponding molecular markers.     

Kinematic evidence for an old stellar halo in the Large Magellanic Cloud

The oldest and most metal-poor Milky Way stars form a kinematically hot halo, which motivates the two major formation scenarios for our galaxy: extended hierarchical accretion and rapid collapse. RR Lyrae stars are excellent tracers of old and metal-poor populations. We measured the kinematics of 43 RR Lyrae stars in the inner regions of the nearby Large Magellanic Cloud (LMC) galaxy. The velocity dispersion equals 53 +/- 10 kilometers per second, which indicates that a kinematically hot metal-poor old halo also exists in the LMC. This result suggests that our galaxy and smaller late-type galaxies such as the LMC have similar early formation histories.     

Identification and sequence characterisation of molecular markers polymorphic between male kiwifruit (Actinidiachinensis var. deliciosa (A. Chev.) A. Chev.) accessions exhibiting different flowering time

Fruit set in kiwifruit is strongly dependent on pollination, which is limited by the lack of efficient male pollen donors, among other factors. We searched for molecular markers that could be polymorphic in relation to flowering time in order to classify male kiwifruit plants to discard those that are not likely to perform as efficient pollen donors. Random amplified polymorphic DNA (RAPD) and modified amplified fragment length polymorphism (AFLP) markers were generated using 41 male kiwifruit plants in two flowering groups, early- and late-flowering males (with respect to the female cultivar ‘Hayward’). One RAPD and nine modified-AFLP markers polymorphic between male plants exhibiting different flowering time were identified, sequenced and analysed in databases. Unweighted pair group method with arithmetic average (UPGMA) clustering and multidimensional scaling showed that these markers could be used to classify the male plants into flowering groups. Analysis of molecular variance (AMOVA) agreed with this classification, showing that most of the genetic variation is found between flowering groups. Sequence analysis based on a database search revealed that the polymorphism PolM contains a 7-nucleotide long element involved in the repression of the phytochrome A gene, that Pol4 is a partial sequence of a phytochrome B gene, and that sequences Pol3, Pol5, Pol7, and Pol9 show high identity with ESTs from kiwifruit buds treated with hydrogen cyanamide. Clustering analysis supported the previous classification of males into flowering groups, making it feasible to predict male plants’ flowering times with respect to the cultivar ‘Hayward’ based upon these molecular markers.     

Spatial heterogeneity provides organic matter refuges for soil microbial activity in the Patagonian steppe, Argentina

In arid and semi-arid ecosystems that are frequently classified as water limited, it is unclear how spatial and temporal variability of vegetation and climate could affect microbially-mediated soil processes. Our objective was to determine how aboveground spatial heterogeneity creates characteristic soil conditions that modulate microbial growth and activity in a semi-arid Patagonian steppe. In particular, we explored how micro-environmental and biogeochemical soil characteristics generated by the native vegetation could control soil β-glucosidase activity. Both life-form (shrubs, grasses, mosses and bare soil) and season exerted strong controls on all measured abiotic (soil temperature and gravimetric soil water content, inorganic nitrogen, pH and total C and N) and biotic (microbial biomass C and β-glucosidase activity) soil characteristics. Partial correlation between β-glucosidase activity and extracellular organic C (EOC) was high across seasons (r = 0.5; P < 0.001) while soil water content did not correlate with soil enzymatic activity (r = 0.09; P > 0.05). We postulate that labile soil carbon rather than water availability functions as a principal limitation of microbial activity in this semi-arid ecosystem, and the distribution of this carbon is, in large part, determined by the patchy distribution of vegetation.     

Cytotoxic and antioxidant activity of 4-methylthio-3-butenyl isothiocyanate from Raphanus sativus L. (Kaiware Daikon) sprouts

There is high current interest in the chemopreventive potential of Brassica vegetables (cruciferae), particularly due to their content in glucosinolates (GL), which upon myrosinase hydrolysis release the corresponding isythiocyanates (ITC). Some ITCs, such as sulforaphane (SFN) from broccoli ( Brassica oleacea italica), have been found to possess anticancer activity through induction of apoptosis in selected cell lines, as well as indirect antioxidant activity through induction of phase II detoxifying enzymes. Japanese daikon ( Raphanus sativus L.) is possibly the vegetable with the highest per capita consumption within the Brassicaceae family. Thanks to a recently improved gram scale production process, it was possible to prepare sufficient amounts of the GL glucoraphasatin (GRH) as well as the corresponding ITC 4-methylthio-3-butenyl isothiocyanate (GRH-ITC) from its sprouts. This paper reports a study on the cytotoxic and apoptotic activities of GRH-ITC compared with the oxidized counterpart 4-methylsulfinyl-3-butenyl isothiocyanate (GRE-ITC) on three human colon carcinoma cell lines (LoVo, HCT-116, and HT-29) together with a detailed kinetic investigation of the direct antioxidant/radical scavenging ability of GRH and GRH-ITC. Both GRH-ITC and GRE-ITC reduced cell proliferation in a dose-dependent manner and induced apoptosis in the three cancer cell lines. The compounds significantly ( p < 0.05) increased Bax and decreased Bcl2 protein expression, as well as producing caspase-9 and PARP-1 cleavage after 3 days of exposure in the three cancer cell lines. GRH-ITC treatment was shown to have no toxicity with regard to normal human lymphocytes (-15 +/- 5%) in comparison with SFN (complete growth inhibition). GRH and GRH-ITC were able to quench the 2,2-diphenyl-1-picrylhydrazyl radical, with second-order rate constants of 14.0 +/- 2.8 and 43.1 +/- 9.5 M(-1) s(-1), respectively (at 298 K in methanol), whereas the corresponding value measured here for the reference antioxidant alpha-tocopherol was 425 +/- 40 M (-1) s (-1). GRH reacted with H2O2 and tert-butyl hydroperoxide in water (pH 7.4) at 37 degrees C, with rate constants of 1.9 +/- 0.3 x 10(-2) and 9.5 +/- 0.3 x 10(-4) M(-1) s (-1) (paralleling recently developed synthetic antioxidants) being quantitatively (>97%) converted to GRE. It is demonstrated that GRH-ITC has interesting antioxidant/radical scavenging properties, associated with a selective cytotoxic/apoptotic activity toward three human colon carcinoma cell lines, and very limited toxicity on normal human T-lymphocytes.     

Determination of nonprotein amino acids and betaines in vegetable oils by flow injection triple-quadrupole tandem mass spectrometry: a screening method for the detection of adulterations of olive oils

A novel screening method using an automated flow injection electrospray ionization tandem mass spectrometry system is proposed for the simultaneous determination of five nonprotein amino acids (β-alanine, alloisoleucine, ornithine, citrulline, pyroglutamic acid) and three betaines (glycine betaine, trigonelline, proline betaine) after derivatization with butanolic HCl. MS/MS experiments were carried out in a triple-quadrupole instrument using multiple reaction monitoring mode in <2 min. The proposed method provided high fingerprinting power to identify the presence of five of the studied compounds in different types of vegetable oils (soybean, sunflower, corn, olive) with LODs at parts per billion levels. The method was validated, and different mixtures of extra virgin olive oil with seed oils were analyzed, achieving the typification for the detection of adulterations in extra virgin olive oils up to 2% w/w. The nonprotein amino acid ornithine was confirmed as a marker for adulteration in the olive oils analyzed.     

Employment of Marine Polysaccharides to Manufacture Functional Biocomposites for Aquaculture Feeding Applications

In this study, polysaccharides of marine origin (agar, alginate and κ-carrageenan) were used to embed nutrients to fabricate biocomposites to be employed in animal feeding. The consistency of biocomposites in water has been evaluated up to 14 days, by several methods: swelling, nutrient release and granulometric analysis. Biocomposites were produced with varying percentages of nutrients (5%–25%) and polysaccharides (1%–2%–3%). All possible biopolymer combinations were tested in order to select those with the best network strength. The best performing biocomposites were those manufactured with agar 2% and nutrients 10%, showing the lowest percentage of water absorption and nutrient release. Biocomposites made of agar 2% and nutrients 10% were the most stable in water and were therefore used to analyze their behavior in water with respect to the release of quercetin, a phenolic compound with demonstrated high antibacterial and antioxidant activities. The leaching of such molecules in water was therefore employed as a further indicator of biocomposite water stability. Altogether, our results confirm the suitability of agar as a binder for biocomposites and provide a positive contribution to aquaculture.