Purification and characterization of an N-acetylglucosamine-binding lectin from Koelreuteria paniculata seeds and its effect on the larval development of Callosobruchus maculatus (Coleoptera: Bruchidae) and Anagasta kuehniella (Lepidoptera: Pyralidae)

This study describes the purification of an N-acetylglucosamine-binding lectin from Koelreuteria paniculata seeds and its effects on the larval development of Callobruchus maculatus and Anagasta kuehniella. The lectin (KpLec) was characterized and isolated by gel filtration, affinity column, and reverse phase chromatography. SDS-PAGE indicated that this lectin is a dimer composed of subunits of 22 and 44 kDa. The N terminus exhibited 40% similarity with Urtiga dioica agglutinin. KpLec was tested for anti-insect activity against C. maculatus and A. kuehniella. With regard to C. maculatus, an artificial diet containing 0.7 and 1% KpLec produced LD(50) and ED(50) value, respectively. However, for A. kuenhiella, an artificial diet containing 0.65% KpLec produced an LD(50), whereas 0.2% KpLec produced an ED(50). The transformation of genes coding for this lectin could be useful in the development of insect resistance in important agricultural crops.     

Ochratoxin A on green coffee: influence of harvest and drying processing procedures

Ochratoxin A is a metabolite produced by Aspergillus and Penicillium species that is nephrotoxic and possibly carcinogenic to humans. The aim of this study was to evaluate ochratoxin A contamination in green coffee obtained by different harvesting and drying operations and from fruits of different ripening stages in order to identify hazards. The research was directed to coffees from the highland area of Rio de Janeiro state (Brazil), which is traded in the domestic market. Twenty-two out of 54 samples contained ochratoxin A at levels ranging from 0.3 to 160 microg/kg. Ochatoxin A contamination levels between different ripe stage fruits were not significant (P > 0.05). "Varri??o" coffee, consisting of fruits that fell from the tree spontaneously and stayed longer on the ground before being harvested, was the most contaminated. Eleven out of 14 samples of varri??o coffee were contaminated. Three out of 10 samples from the northwestern region of the state were positive for ochratoxin at levels ranging from 10.1 to 592 microg/kg. The contaminated samples had in common the fact that they were harvested directly from the soil.     

A chemometric approach to the comparison of different sample treatments for metals determination by atomic absorption spectroscopy in aceto Balsamico tradizionale di Modena

A comparison of different digestion procedures has been carried out for the analysis of metal concentration in samples of vinegars and Aceto Balsamico Tradizionale of Modena (ABTM) coming from an unique barrel set. In particular, classical wet, dry ashing, and closed vessel microwave digestion procedure have been utilized and compared for each investigated species. In a few cases, direct metal determination on ABTM (without treatment procedure) is proposed as possible alternative to sample manipulation. Flame atomic absorption spectrometry was used for the quantification of iron and zinc, while graphite furnace atomic absorption spectrometry was used for all the other elements (i.e., chromium, manganese, cobalt, nickel, copper, cadmium, and lead). The comparison among the different sample treatments was carried out by the use of statistical and chemometric tools. In particular, principal component analysis and ANOVA approaches were used to discriminate between the diverse analytical methods. Furthermore, for all the dissolving techniques, the analytical metal recovery was always evaluated by the application of the recovery function on the same sample matrix. In general, the recoveries were fairly good, ranging from 90 to 103%, except for Cd and Pb with dry ashing, which showed recovery values close to 55% and 67%, respectively. As regards the metals concentration of the investigated samples, the experimental data reveal for some species the presence of concentration slightly over the legal limit fixed for wine and wine vinegar.     

Effect of processing on the antioxidant vitamins and antioxidant capacity of Vigna sinensis Var. Carilla

Cowpea (Vigna sinensis L. var. Carilla) flours obtained by fermentation with inoculum Lactobacillus plantarum (PF) or with the natural microorganisms present in the flour (NF) and subsequent heat treatment in an autoclave were prepared to study the effect of fermentation on the antioxidant vitamin content and on the antioxidant capacity. Bacterial counts and pH values, vitamins C and E, carotenoids, glutathione (GSH), superoxide dismutase-like activity (SOD-like activity), peroxyl radical-trapping capacity (PRTC), lipid peroxidation in unilamillar liposomes, and Trolox equivalent antioxidant capacity (TEAC) were evaluated in raw and processed cowpea flours. gamma-Tocopherol and delta-tocopherol were found in raw cowpea, whereas vitamin C and carotenoids were not detected. An increase in the vitamin E activity was observed in PF, whereas vitamin C and carotenoids were not detected in fermented cowpea flours. Fermentation or heat treatment in an autoclave after fermentation produced processed cowpea flours with lower PRTC, glutathione content, and SOD-like activity than those of the raw seeds. However, those processes increased the capacity to inhibit the lipid peroxidation in unilamellar lipoposomes and TEAC. According to the results obtained in this study, the fermentation of cowpeas (naturally or with L. plantarum) and fermentation and subsequent heat treatment in an autoclave are good processes to obtain functional cowpea flours having higher antioxidant capacity than the raw legume.     

Induction of de novo volatile terpene biosynthesis via cytosolic and plastidial pathways by methyl jasmonate in foliage of Vitis vinifera L

The terpene biosynthesis in leaves of Vitis vinifera L. cv. Morio Muskat was studied using methyl jasmonate to induce defensive responses in vivo. The experiments demonstrated the strong activation of the de novo biosynthesis of terpenoids via the octadecanoid-signaling cascade and release of the compounds to the gas phase. Feeding experiments with [5,5-(2)H(2)]-1-deoxy-d-xylulose and [5,5-(2)H(2)]mevalonic acid lactone allowed the investigation of the dynamic allocation of resources via the mevalonic acid and 1-deoxy-d-xylulose/2-C-methyl-d-erythritol 4-phosphate (DOXP/MEP) pathway under induced conditions and after treatment with the specific inhibitors mevastatin and fosmidomycin. The experiments reveal that monoterpenes are almost exclusively synthesized via the DOXP/MEP pathway, whereas sesquiterpenes are generated via both pathways at approximately equal rates. The biosynthesis of the homoterpene (E)-4,8-dimethyl-1,3,7-nonatriene was not affected by mevastatine or fosmidomycin.     

Simultaneous and sensitive detection of three foodborne pathogens by multiplex PCR, capillary gel electrophoresis, and laser-induced fluorescence

The simultaneous detection of Staphylococcus aureus, Listeria monocytogenes, and Salmonella spp. has been approached by a new multiplex PCR-based procedure followed by capillary gel electrophoresis with laser-induced fluorescence detection (multiplex-PCR-CGE-LIF). As compared to slab gel electrophoresis, the use of CGE-LIF improved from 10- to 1000-fold the sensitivity of the multiplex PCR analysis, allowing the detection of 2.6 x 10(3) cfu mL(-1) of S. aureus, 570 cfu mL(-1) of L. monocytogenes, and 790 cfu mL(-1) of Salmonella in artificially inoculated food, without enrichment. Following 6 h of enrichment, as low as 260, 79, and 57 cfu mL(-1) of S. aureus, L. monocytogenes, and Salmonella, respectively, were detected. The CGE-LIF method is shown to be reproducible, providing relative standard deviation (RSD) values lower than 0.8% for analysis time and lower than 5.8% for peak areas. The multiplex-PCR-CGE-LIF proved a powerful analytical tool to detect various food pathogens simultaneously in a fast, reproducible, and sensitive way.     

Vitamin E composition of walnuts (Juglans regia L.): a 3-year comparative study of different cultivars

The tocopherol and tocotrienol composition of walnuts (Juglans regia L.) was determined for nine cultivars (cvs. Arco, Franquette, Hartley, Lara, Marbot, Mayette, Mellanaise, Parisienne, and Rego). Walnuts were harvested over three consecutive years from two different geographical origins (Bragan?a and Coimbra, Portugal), for a total of 26 samples. The methodology employed was a normal-phase high-performance liquid chromatography coupled to a series arrangement of a diode array detector followed by a fluorescence detector, allowing the simultaneous analysis of all tocopherols and tocotrienols. The analyses showed that all samples presented a similar qualitative profile composed of five compounds: alpha-tocopherol, beta-tocopherol, gamma-tocopherol, delta-tocopherol, and gamma-tocotrienol. gamma-Tocopherol was the major compound in all samples, ranging from 172.6 to 262.0 mg/kg, followed by alpha- and delta-tocopherols, ranging from 8.7 to 16.6 mg/kg and from 8.2 to 16.9 mg/kg, respectively. Multivariate analysis of the data obtained showed the existence of significant differences in composition among cultivars. These differences were also significant when cultivars were grouped by year of production, showing that besides genetic factors, the vitamin E composition was influenced by environmental factors.     

Short-term survival and dispersal of translocated European wild rabbits. Improving the release protocol

Translocation of European wild rabbit Oryctolagus cuniculus L. is one of the most frequent management tools to increase rabbit density in Spain, both as prey of several predators that are threatened with extinction and for sport hunting. Nevertheless the elevated short-term mortality by predation makes translocations unsuccessful and increases their biological cost. Information on the factors affecting the short-term survival and dispersal of translocated rabbits is required to improve release management and increase performance of translocated rabbits, and to avoid the use of non-selective lethal methods for predator control. In this study we tested electric fencing and night-shooting as alternative to traditional release protocols, and the effects of vegetation cover and warren fencing on short-term survival and dispersal of rabbits. Night shooting performed during the first nights after release increased significantly the survival of rabbits, by hindering the activity of carnivores in the release area. The use of an electric fence enclosure also increased the performance of rabbits, but was not efficient to constraint rabbit dispersal. Rabbits released in areas with low vegetation cover showed higher mortality and dispersal distances than rabbits released in high cover areas. Warren fencing decreased both the dispersal of rabbits and the adverse impact of predation in low cover areas, but had no effect in high cover areas. Selection of high cover areas or warren fencing in low cover areas seem to be the most advantageous release conditions to decrease the short-term predation impact, reducing the biological cost of rabbit translocations and the risks for endangered predators derived from the use of traditional predator control practices during translocations.