Diversity of native rice (Oryza Poaceae:) species of Costa Rica

We found several populations of wild Oryza species in the lowlands of Costa Rica. The plants showed extensive morphological variation, suggesting the presence of several species. In the morphologic study, 33 traits were scored for plants of all the species. A principal component analysis revealed the significant morphological separation of the different species. The analyses indicated that there are three species, O. grandi­glumis, O. latifolia and O. glumaepatula. Two putative hybrid types were found, both significantly differing in their morphology from the known species and intermediate at several traits. O. grandi­glumis is a new record for Costa Rican flora. Its main population is located in Caño Negro Wildlife Refuge, Los Chiles, Alajuela. O. latifolia is distributed throughout the lowlands of the country and the plants of the Atlantic slope are significantly bigger in general habit than those of the Guanacaste area. During this study a population of O. glumaepatula of hundreds of thousands of plants was discovered in the Medio Queso River wetland, Los Chiles, Alajuela. This population is the most important source of genes for cultivar's improvement from the primary gene pool of rice in Costa Rica. The small ligule and the wide flag leaf characteristic of the two CCDD species separated them from the AA diploid O. glumaepatula. Seed size, ligule size, number of branches in the panicle, plant height and sterile lemma length are all bigger in O. grandi­glumis, and influenced the second factor that separated the CCDD species in two discrete clusters. The species found offer great possibilities for the improvement of rice cultivars and they should be thoroughly studied and appropriately protected.     

Clonal production and organization of inhibitory interneurons in the neocortex

The neocortex contains excitatory neurons and inhibitory interneurons. Clones of neocortical excitatory neurons originating from the same progenitor cell are spatially organized and contribute to the formation of functional microcircuits. In contrast, relatively little is known about the production and organization of neocortical inhibitory interneurons. We found that neocortical inhibitory interneurons were produced as spatially organized clonal units in the developing ventral telencephalon. Furthermore, clonally related interneurons did not randomly disperse but formed spatially isolated clusters in the neocortex. Individual clonal clusters consisting of interneurons expressing the same or distinct neurochemical markers exhibited clear vertical or horizontal organization. These results suggest that the lineage relationship plays a pivotal role in the organization of inhibitory interneurons in the neocortex.     

Behavioural response of a trophic specialist, the Iberian lynx, to supplementary food: Patterns of food use and implications for conservation

Prey scarcity compromises population survival, especially for specialist predators. Supplementary feeding is a management tool that can be applied to reverse the decline of food-limited populations. We analyse how a population of Iberian lynx, a threatened food specialist, initially reacted to, and subsequently used, supplementary food. Twenty-seven feeding stations (FS) with domestic rabbits were placed in the Doñana Biological Reserve, SW Spain, between 2002 and 2005. We recorded lynx tracks inside and around stations to analyse spatio-temporal patterns of use, as well as the performance of several station designs. Iberian lynx used 62% of the FS installed, and consumed most of the supplied food. All station designs were used and food provision apparently covered the energetic needs of the lynx inhabiting the reserve. There was spatial aggregation in the use of the FS. Fourteen weeks were needed on average for lynx to become familiar with feeding stations and making a regular use of the supplementary food. Seasonal variations in consumption frequency appeared to be modulated by fluctuations of wild rabbit numbers throughout its annual cycle as well as by variation in energy demand of breeding females. The Iberian lynx responded positively to our supplementary feeding programme. We show that this technique allows the persistence of lynx populations during long periods in areas where wild rabbits are extremely scarce. Supplementary food may be used to sustain lynx whenever rabbit populations need recovery, as well as in the context of lynx restocking or reintroduction programmes.     

Cytoprotection by neutral fraction of tannat red wine against oxidative stress-induced cell death

Some of the beneficial effects of the Mediterranean diet on human pathologies have been attributed to red wine polyphenols. It has been postulated that the antioxidant activity of the latter would be also responsible for the cytoprotective capacity of red wine that has been reported in a few papers. Nevertheless, red wine shows a complex composition, and the active fraction is not known yet. In this context, the protective capacity of total lyophilized extracts of red wine and anthocyanin, neutral, or acidic fractions, was explored in PC12 cells in culture after a hydrogen peroxide insult. Although all fractions showed high antioxidant activity, only the neutral fraction was cytoprotective. The analysis of this active fraction showed that it was rich in the aglycons quercetin and myricetin as well as the glycosides of kaempferol, isorhamnetin, epicatechin, and catechin, some of which are known to be cytoprotective. This is the first paper to reveal the active fraction of total wine responsible of its cytoprotection.     

Predation of artificial nests in a fragmented landscape in the tropical region of Los Tuxtlas, Mexico

Predation rates of artificial nests were investigated in a fragmented landscape in the lowlands of Los Tuxtlas in southern Mexico. Hen and plasticine eggs were used to assess predation pressure in four habitats: the interior of forest fragments, the forest-pasture edge, corridors of residual forest vegetation and linear strips of live fences across pastures. Three sites per habitat were used in three experimental trials. Hen and plasticine ground nests with three eggs each were alternated every 50 m along transects at each site. Predation rates on each type of nest were monitored for 9 days. Survey of potential avian and mammalian potential nest predators were conducted at each site prior to the experimental trails. Readings of amount of light illuminating the ground were taken by each nest at each site to assess exposure of nests. In general, average predation rates were significantly higher for both hen and plasticine nests in the forest-pasture edge and in the corridors than in the interior of the forest fragments. While birds and mammals were the principal predators on hen eggs in the forests, mammals were responsible for the majority (?70%) of eggs damaged at the other habitats. Surveys of potential nest predators showed that avian and mammalian potential nest predators were significantly more common at the forest-pasture edges and at the other habitats than in the forests. Readings of light reaching the ground suggest that concealment of nests by the vegetation may play an important role in predation risk. Our results are consistent with reports from other Neotropical rainforests indicating an increase of artificial nest predation pressures from forest interior to open habitats. Restoration of forest fragments, allowing the vegetation to grow along the forest-pasture edge and the planting of arboreal crops at the forest-pasture edges may be measures that could increase cover and nest protection.     

Countercurrent supercritical fluid extraction and fractionation of high-added-value compounds from a hexane extract of olive leaves

Countercurrent supercritical fluid extraction (CC-SFE) at a pilot scale plant was used for fractionation of high-added-value products from a raw extract of olive leaves in hexane. Compounds found in the raw extract were waxes, hydrocarbons, squalene, beta-carotene, triglycerides, alpha-tocopherol, beta-sitosterol, and alcohols. The CC-SFE extraction process was investigated according to a 2(3) full factorial experimental design using the following variables and ranges: extraction pressure, 75-200 bar; extraction temperature, 35-50 degrees C; and ethanol as modifier, 0-10%. Data were analyzed in terms of extraction yield, enrichment, recovery, and selectivity. Higher extraction yields were attained at 200 bar. For most of the compounds analyzed enrichment was attained at the same conditions, that is, 75 bar, 35 degrees C, and 10% ethanol. Hydrocarbons were usually recovered in the separators, whereas waxes and alpha-tocopherol remain in the raffinate. Selectivity data reveal that alpha-tocopherol is the most easily separable compound. The influence of the experimental factors on the recovery of all the compounds was studied by means of regression models. The best fitted model was attained for beta-sitosterol, with R2 = 99.25%.     

Silvopastoral systems improve carbon stocks at livestock ranches in Tabasco,Mexico

Silvopastoral systems have great potential for storing carbon because of carbon assimilation in tree woody biomass, carbon input through litterfall and below-ground carbon turnover. In this study, we quantified and compared the carbon stocks at livestock ranches in Tabasco, Mexico, containing either scattered trees in grazing pastures (STP) or grass monocultures. Sampling plots were randomly established at each ranch where the above- and below-ground carbon stocks, carbon input from litterfall, grass production and arboreal biomass growth were measured. We found that silvopastoral systems stored an average of 257.45 Mg ha⁻¹ of soil organic carbon (SOC) compared to 119.17 Mg SOC ha⁻¹ at grass monoculture ranches (to 30 cm depth); silvopastoral systems also stored 44.64 Mg C ha⁻¹ in wood biomass; and, grass monocultures had greater cumulative grass biomass production. Overall, it is concluded that livestock ranches in Tabasco, Mexico, with scattered trees in grazing pastures stored 58.8% more carbon than those grass monocultures, with carbon stocks of 327.01 Mg C ha⁻¹and 134.47 Mg C ha⁻¹, respectively. The results are useful for land management decision making for sustainable livestock systems framed in the Sustainable Development Goals (SDGs).     

Ultrasensitive detection of genetically modified maize DNA by capillary gel electrophoresis with laser-induced fluorescence using different fluorescent intercalating dyes

In this work, four different fluorescent intercalating dyes are compared for the ultrasensitive CGE-LIF detection of DNA from transgenic maize in flours. The fluorescent intercalating dyes compared are YOPRO-1, SYBR-Green-I, Ethidium bromide (EthBr), and EnhanCE. For all the four dyes optimum concentrations are established, and efficient separations of DNA fragments ranging in size from 80 to 1000 bp are obtained. The comparative study demonstrates that SYBR-Green-I and YOPRO-1 provide better limits of detection (LODs) than EnhanCE or EthBr (i.e., LODs are, respectively, 700, 1000, 11300, and 97400 zmol, calculated for a 200-bp DNA fragment). Separations using YOPRO-1 are faster than those using SYBR-Green-I (30 min vs 47 min for the analysis of the 80-1000 bp DNA fragments). Also, separations using YOPRO-1 are more efficient than those using SYBR-Green-I (e.g., 2.4 x 10(6) plates/m vs 1.6 x 10(6) plates/m, respectively, calculated for the 200-bp fragment). Also, buffer depletion and cost per analysis are worse with SYBR-Green-I than with YOPRO-1. Therefore, YOPRO-1 was selected as the preferred intercalating dye. Using this fluorescent compound, analysis time reproducibility for the CGE-LIF separation of the DNA fragments is determined to be better than 1.7% (% RSD, n = 10) within the same day, and better than 1.9% (% RSD, n = 30) for three different days. Moreover, the fluorescence signal obtained using this dye is shown to vary linearly with the DNA concentration in the range studied, i.e., 1-500 ng/microL. It is demonstrated that by using this method 0.01% of transgenic maize can be detected in flour by direct injection of the PCR-amplified sample.     

Characterization of protein fractions from Bt-transgenic and non-transgenic maize varieties using perfusion and monolithic RP-HPLC. Maize differentiation by multivariate analysis

Protein fractions from transgenic Bt and non-transgenic maize varieties, extracted by the Osborne solvent fraction procedure, were characterized for the first time by perfusion and monolithic RP-HPLC in very short analysis times. Albumins and globulins from different transgenic Bt maizes as well as from their non-transgenic isogenic varieties were eluted in four peaks using perfusion RP-HPLC, whereas prolamins and glutelins were separated in seven peaks. Monolithic RP-HPLC enabled the separation of maize proteins in a large number of peaks showing 6 and 10 main peaks for albumins and globulins, respectively. Prolamins migrated at retention times higher than 5 min as seven peaks, whereas glutelins were separated in three main peaks appearing at retention times higher than 6.0 min. Moreover, chromatograms of the whole protein extracts showed 8 and 11 components for perfusion and monolithic RP-HPLC, respectively. A comparison of the chromatograms of the whole protein extracts relative to transgenic and non-transgenic varieties evidenced quantitative differences on the percentages of area, mainly for peaks 2 and 3 by perfusion RP-HPLC and for peaks 3 and 7 by monolithic RP-HPLC. A discriminant analysis based on these proteic profiles was carried out to classify and predict transgenic Bt maize lines, achieving 100% correct classification using perfusion RP-HPLC.