Conversion of the mycotoxin patulin to the less toxic desoxypatulinic acid by the biocontrol yeast Rhodosporidium kratochvilovae strain LS11

The infection of stored apples by the fungus Penicillium expansum causes the contamination of fruits and fruit-derived products with the mycotoxin patulin, which is a major issue in food safety. Fungal attack can be prevented by beneficial microorganisms, so-called biocontrol agents. Previous time-course thin layer chromatography analyses showed that the aerobic incubation of patulin with the biocontrol yeast Rhodosporidium kratochvilovae strain LS11 leads to the disappearance of the mycotoxin spot and the parallel emergence of two new spots, one of which disappears over time. In this work, we analyzed the biodegradation of patulin effected by LS11 through HPLC. The more stable of the two compounds was purified and characterized by nuclear magnetic resonance as desoxypatulinic acid, whose formation was also quantitated in patulin degradation experiments. After R. kratochvilovae LS11 had been incubated in the presence of (13)C-labeled patulin, label was traced to desoxypatulinic acid, thus proving that this compound derives from the metabolization of patulin by the yeast. Desoxypatulinic acid was much less toxic than patulin to human lymphocytes and, in contrast to patulin, did not react in vitro with the thiol-bearing tripeptide glutathione. The lower toxicity of desoxypatulinic acid is proposed to be a consequence of the hydrolysis of the lactone ring and the loss of functional groups that react with thiol groups. The formation of desoxypatulinic acid from patulin represents a novel biodegradation pathway that is also a detoxification process.     

Factors affecting the synergy of thiabendazole, sodium bicarbonate, and heat to control postharvest green mold of citrus fruit

The efficacy of thiabendazole (TBZ) to control postharvest decay caused by Penicillium digitatum of citrus fruit can be enhanced by co-application with sodium bicarbonate (SBC) and/or heat treatment. The impact of these treatments was investigated in citrus fruit, as a function of TBZ and SBC concentration and temperature, and were related to the amount of TBZ residues in fruit (total residues), in fruit surface, in the cuticular wax, and in the inner fruit. The residue levels of TBZ were determined in 'Valencia' oranges following a 1 min dip in an aqueous mixture of SBC at 0.5, 1, or 2 wt %/vol and TBZ at 600 or 400 mg/L (active ingredient, a.i.) at 20 or 40 degrees C and after 0 and 20 days at 17 degrees C and 90% relative humidity. The influence of SBC and heat on the TBZ residue concentration on the fruit surface, in cuticular wax, and on the inner cuticle tissue was determined in 'Salustiana' oranges after a 1 or 3 min dip in TBZ alone at 600 mg/L and 20 or 50 degrees C or for 1 min in TBZ at 600 mg/L and SBC at 2% and 20 degrees C. The efficacy of heat treatments with water, SBC, and TBZ, applied separately or in combination, was investigated on artificially inoculated 'Nova' mandarins and 'Valencia' oranges for the control of postharvest green mold caused by a TBZ-sensitive (TBZ-s) or TBZ-resistant (TBZ-r) isolate of P. digitatum. The residue levels of TBZ in fruit, evaluated as total residues, were not affected by the co-application of SBC in most samples. While TBZ residues in the fruit surface were not significantly affected by the dip temperature or by co-application of SBC, the rates of diffusion and penetration of TBZ into cuticular wax markedly increased in the presence of SBC or when TBZ was applied in combination with heat. TBZ residues in the inner tissue of fruits treated at 20 degrees C were not dependent upon the dip time or by the presence of SBC and were similar to those found in fruit treated with TBZ at 50 degrees C for 1 min, whereas significantly higher values were recorded in samples treated with TBZ at 50 degrees C for 3 min. When TBZ at 600 mg/L and 20 degrees C was applied in the presence of SBC at concentrations of 1-2 or 0.5-2%, it effectively reduced decay caused by the TBZ-resistant isolate of green mold in 'Nova' mandarins and 'Valencia' oranges. This treatment was also significantly more effective than TBZ alone to control green mold caused by a TBZ-s isolate in 'Valencia' oranges. The combination with SBC and mild heat (40 degrees C) and TBZ at 400 mg/L generally improved the control of a TBZ-r isolate of green mold with respect to the combined treatment at 20 degrees C. TBZ efficacy was also improved when applied at reduced rates (200 mg/L) and 50 degrees C, significantly suppressing green mold caused by a TBZ-s isolate of P. digitatum and effectively controlling a TBZ-r isolate. The rate of weight loss of 'Valencia' oranges was significantly increased by SBC treatment and was positively dependent upon the concentration of SBC used in the treatment, while the temperature of the treatment solution had little influence on later weight loss.     

One- and two-dimensional electrophoretic identification of IgE-binding polypeptides of Lupinus albus and other legume seeds

The prevalence of food allergies in the world population requires integrated approaches to identify new potential allergens, especially those of plant origin. The aim of this work was the allergen in vitro analysis of Lupinus albus seed proteome, a promising food protein source, and the assessment of IgE cross-reactivities with other more diffused legume species. A combination of one- and two-dimensional gel electrophoresis and immunoblotting analyses with specific IgGs for band identification and lupin-sensitized patients' circulating IgEs for allergenicity studies has been used. Two lupin proteins, namely, conglutin gamma and 11S globulin basic subunits, strongly reacted with all patients' sera. Also, cross-reactivities with the homologous polypeptides of other legume species were observed. Otherwise, no reaction at all was detected with a 2S-type lupin protein. This global electrophoretic approach has allowed the identification of a new potential lupin allergen and confirmed the cross-reactivity among the legume 11S globulin basic subunits.     

Residue levels and effectiveness of pyrimethanil vs imazalil when using heated postharvest dip treatments for control of Penicillium decay on citrus fruit

The influence of fungicide concentration and treatment temperature on residue levels of pyrimethanil (PYR) in comparison with the commonly used fungicide imazalil (IMZ) was investigated in orange fruits following postharvest dip treatments. The dissipation rate of PYR residues was recorded as a function of storage conditions. The fungicide efficacy against green and blue molds caused by Penicillium digitatum and Penicillium italicum, respectively, was evaluated on different citrus varieties following the fungicide application at 20 or 50 degrees C. Residue levels of PYR in Salustiana oranges were significantly correlated with the fungicide dosage, but residue concentrations were notably higher (ca. 13-19-fold) after treatment at 50 degrees C as compared to treatments at 20 degrees C. After treatment at temperatures ranging from 20 to 60 degrees C, PYR and IMZ residues in Salustiana oranges were significantly correlated with dip temperatures. Dissipation rates of PYR during storage were negligible in both Salustiana and Tarocco oranges. Results obtained on wounded, noninoculated Miho satsumas revealed that when treatments were performed at 50 degrees C, PYR or IMZ concentrations needed to achieve the complete control of decay were 8- and 16-fold less than by treatment at 20 degrees C. When fruits were inoculated with either P. digitatum or P. italicum, the application of 400 mg L(-1) PYR at 20 degrees C or 100 mg L(-1) PYR at 50 degrees C similarly reduced green and blue mold development. These results were corroborated by storage trials on Marsh grapefruits and Tarocco oranges. The lowest concentration of PYR required to achieve almost total protection of the fruit against decay accounted for 100 mg L(-1) at 50 degrees C and 400 mg L(-1) at 20 degrees C, respectively. Treatments did not affect fruit external appearance, flavor, and taste. It is concluded that postharvest PYR treatment represents an effective option to control green and blue mold in citrus fruit and that integration of fungicide applications and hot water dips may reduce the possibility of selecting fungicide-resistant populations of the pathogen, as a consequence of increased effectiveness of the treatment.     

Determination of free and bound phenolic compounds in buckwheat spaghetti by RP-HPLC-ESI-TOF-MS: effect of thermal processing from farm to fork

Nowadays there is considerable interest in the consumption of alternative crops as potential recipes for gluten-free products production. Therefore, the use of buckwheat for the production of gluten-free pasta has been investigated in the present study. RP-HPLC-ESI-TOF-MS has been applied for the separation and characterization of free and bound phenolic compounds in buckwheat flour and buckwheat spaghetti. Thus, 32 free and 24 bound phenolic compounds in buckwheat flour and spaghetti have been characterized and quantified. To the authors' knowledge, protochatechuic-4-O-glucoside acid and procyanidin A have been detected in buckwheat for the first time. The results have demonstrated a decrease of total free phenolic compounds from farm to fork (from flour to cooked spaghetti) of about 74.5%, with a range between 55.3 and 100%, for individual compounds. The decrease in bound phenols was 80.9%, with a range between 46.2 and 100%. The spaghetti-making process and the cooking caused losses of 46.1 and 49.4% of total phenolic compounds, respectively. Of the total phenolic compounds present in dried spaghetti, 11.6% were dissolved in water after cooking.     

A giant European dinosaur and a new sauropod clade

Fossils of a giant sauropod dinosaur, Turiasaurus riodevensis, have been recovered from terrestrial deposits of the Villar del Arzobispo Formation (Jurassic-Cretaceous boundary) of Riodeva (Teruel Province, Spain). Its humerus length (1790 millimeters) and estimated mass (40 to 48 metric tons) indicate that it may have been the most massive terrestrial animal in Europe and one of the largest in the world. Phylogenetic analysis indicates that the fossil represents a member of a hitherto unrecognized group of primitive European eusauropods that evolved in the Jurassic.     

Study of conifer nitrogen nutrition using hydroponic cultures: Application to fertigation systems

Nitrogen (N) optimization of a nutrient solution for ornamental conifers was obtained with two different experiments in hydroponic cultures: First, by using three different NO₃ ^‐/NH₄ ⁺ ratios (60/40, 55/45, and 40/60 in percentages of the total N supplied); and, secondly by testing three total N levels (3.7; 4.7 and 5.5 mmol L^‐1). Best growth was obtained with a NO₃ ^‐/NH₄ ⁺ ratio of 55/ 45 and a total N level of 3.7 mmol L^‐1. With these experiments, reference levels of foliar concentrations of the macronutrients N, phosphorus (P), potassium (K), calcium (Ca), magnesium (Mg), and the biochemical indices, such as chlorophyll and starch levels, were obtained with the treatments corresponding to the plants with the higher growth. In the course of a growth cycle, a substrate assay with two different pot mixes (moss peat plus perlite and black peat plus sepiolite 60/40 ‐% v/v‐) was carried out by using the best N ratios and doses in nutrient solutions as obtained in hydroponics. The results indicated that the same N fertilization in fertigation systems changes depending on the different physicochemical properties of the substrates used; in this case, depending on the different physical properties of the two substrates. By applying DRIS to perform nutritional diagnosis, it is possible to find nutritional limitations to plant growth, but not additional factors, such as water‐air relationships in the growth media.     

Application of a Balanced Scorecard System for Supporting Decision-Making in Contaminated Sites Remediation

This work introduces the development and application of a multi-goal evaluation procedure for assessing sets of remedial options that have previously been found technically suitable for managing a certain contaminated site. The construction of this decision support tool, in the form of a balanced scorecard, has followed a multi-attribute conception of the contaminated sites problem, identifying the most critical aspects considered by all stakeholders involved in contaminated sites management decisions. The developed tool has been checked in its formulation and application against an export benchmark of an Austrian remediation project. The performed evaluation allowed to identify and suggest a remedial option that best fitted the expectations of all stakeholders involved in the project. Furthermore, through the accomplishment of a sensitivity analysis it has been demonstrated that the system is stable and robust in terms of the chosen elicitation schemes, and that the preferences of the different stakeholders are influencing factors on the final result. The results and conclusions of this work present the benefits of implementing the developed balanced scorecard as a tool for supporting decision making in contaminated sites remediation projects.     

Simple and inexpensive flow L-glutamate determination using pumpkin tissue

This work refers to a very easy to implementate flow injection system with potentiometric detection for l-glutamate determination in food samples. The proposed procedure is based on measurement of carbon dioxide produced by decarboxylation of l-glutamate catalyzed by l-glutamate decarboxylase (E.C. 4.1.1.1.5) from Cucurbita maxima (pumpkin). The FI potentiometric system includes an enzymatic reactor with a length of 8 cm and thickness of 5 mm packed with 200 mg of a C. maxima outer layer cut in to small pieces. The proposed procedure allowed l-glutamate determinations in the concentration interval of 10-100 mmol L(-1) for an injected sample volume of 50 microL. A phosphate buffer (0.1 mol L(-1), pH 5.5) solution flowing at 1.4 mL min(-1) was used as the carrier solution in the system. The results obtained in the analysis of food samples revealed a relative error lower than 5% when compared with those provided by the spectrophometric reference procedure. The immobilized reactor retained its initial activity for 21 days. It was possible to measure 40 samples/h with the flow system proposed.     

Rotenone and rotenoids in cubè resins, formulations, and residues on olives

Rotenone and rotenoids (deguelin, beta-rotenolone (12a beta-hydroxyrotenone), tephrosin (12a beta-hydroxydeguelin), 12a alpha-hydroxyrotenone, and dehydrorotenone) were determined in cubè resins and formulations. Cubè resins from Lonchocarpus contain large quantities of deguelin (ca. 21.2%) and smaller quantities of tephrosin (ca. 3.5%) and beta-rotenolone (ca. 3.0%). The composition of commercial formulations may present very different rotenoid contents depending on the extracts used to prepare them. Because these rotenoids also present insecticide activity, the efficacy of these formulations may be very different. The storage stability and photodegradation of some rotenone formulations were studied. Rotenone and rotenoids are very sensitive to solar radiation, which degrades them rapidly, with half-lives in the order of a few tens of minutes. Some formulations show greater disappearance rates than that of cubè resin, indicating that not much attention has been paid to protecting the active ingredients from photodegradation in the formulation. A study on the residues on olives was also carried out to assess not only the rotenone content, but also that of the main rotenoids. At harvest, the residues of deguelin, tephrosin, and beta-rotenolone were 0.10, 0.06, and 0.10 mg/kg, respectively, very similar to rotenone (0.08 mg/kg), and though a few data indicate similar acute toxicity values for deguelin, only rotenone is taken into consideration in the legal determination of the residue.