Identification and quantification of phenolic compounds in grapes by HPLC-PDA-ESI-MS on a semimicro separation scale

Reversed phase high performance liquid chromatography (RP-HPLC) on a semimicro separation scale was employed to develop a straightforward method for the simultaneous separation, identification, and quantification of phenolic compounds occurring in whole berries of Vitis vinifera, which comprise phenolic acids, flavonols, catechins, stilbenes, and anthocyanins. A C-18 narrow bore column of 150 x 2.0 mm I.D. and a semimicro photodiode array detector (PDA) cell of 2.5 microL, in conjunction with a mass spectrometry detector equipped with an electrospray ionization source (ESI-MS) to confirm peak identification, were employed. The C-18 narrow bore column was eluted by a multisegment gradient of increasing concentration of acetonitrile in water-formic acid solution that was optimized on the basis of the results of a study carried out to evaluate the influence of mobile phase composition and gradient shape on separation performance and detection sensitivity by ESI-MS. The identification of individual phenolic compounds was performed on the basis of their retention times and both UV-visible and mass spectra, acquired by a mass spectrometer (MS) equipped with an electrospray ionization (ESI) source, employed in conjunction with the PDA detector. Libraries comprising retention times, UV-visible, and mass spectra for major phenolic compounds expected in grape berries were made by subjecting solutions of each phenolic standard to the optimized RP-HPLC method. Quantification of individual compounds was performed by the external standard method using a six point regression graph of the UV-visible absorption data collected at the wavelength of maximum absorbance of each analyte determined by the PDA spectra. The RP-HPLC method was validated in terms of linearity of calibration graphs, limits of detection, limits of quantification, repeatability, and accuracy, which was evaluated by a recovery study. The developed method was successfully applied to identify the phenolic compounds occurring in the whole berries of nine red and one white grape of different varieties of Vitis vinifera, comprising some autochthonous varieties of south Italy such as Aglianico, Malvasia Nera, Uva di Troia, Negroamaro, Primitivo, and Susumaniello. Large differences in the content of phenolic compounds was found in the investigated grape varieties. As expected, only glycosilated flavonols were quantified, and the total amount of these compounds was higher in the whole berries of red grapes than in the white Moscato, where the most abundant phenolic compound was quercetin 3-O-glucoside. In almost all samples, the most and least abundant anthocyanins were malvidin 3-O-glucoside and cyanidin 3-O-glucoside, respectively, with the exception of Uva di Troia where the least abundant anthocyanin was delphinidin 3-O-glucoside.     

The importance of demographic niches to tree diversity

Most ecological hypotheses about species coexistence hinge on species differences, but quantifying trait differences across species in diverse communities is often unfeasible. We examined the variation of demographic traits using a global tropical forest data set covering 4500 species in 10 large-scale tree inventories. With a hierarchical Bayesian approach, we quantified the distribution of mortality and growth rates of all tree species at each site. This allowed us to test the prediction that demographic differences facilitate species richness, as suggested by the theory that a tradeoff between high growth and high survival allows species to coexist. Contrary to the prediction, the most diverse forests had the least demographic variation. Although demographic differences may foster coexistence, they do not explain any of the 16-fold variation in tree species richness observed across the tropics.     

The 70-kDa Heat-shock Protein as a Potential Biomarker of Quality of the Parapenaeus longirostris Shrimp Flesh

The deep-water rose shrimp, Parapenaeus longirostris, is a target species of the Mediterranean fisheries mostly caught by trawlers offshore, processed, and frozen on board. The effects of thawing on shrimp muscle exudate collected at 0, 1, 2, 3 days after thawing were investigated. In total, 70-kDa heat-shock protein (Hsp70), alpha (α)-enolase, and manganese superoxide dismutase (Mn-SOD) were selected as metabolic and stress-related proteins and analyzed by immunoblotting on exudates. Data were compared for the amount of exudates collected and the pH values. Among the investigated proteins, only the Hsp70 levels showed a decrease related to the post-thawing period and correlated with both the significant increase of the exudate amount and the pH values. These data strongly suggest the potential use of Hsp70 as an early predictive biomarker for quality of the P. longirostris shrimp after thawing.     

Identification of candidate genes for paratuberculosis resistance in the native Italian Garfagnina goat breed

Paratuberculosis disease is a chronic bacterial disease infection of ruminants of global relevance, caused by MAP (Mycobacterium avium subsp. paratuberculosis). The present study was conducted on the Garfagnina goat breed that is an Italian native goat population registered on the Tuscan regional repertory of genetic resources at risk of extinction. Forty-eight adult goats (27 serologically positive to MAP-positive and 21 serologically negative to MAP-negative) belonging to a single flock that had experienced annual mortalities due to MAP infection were identified and genotyped with the Illumina GoatSNP60 BeadChip. Diagnosis was achieved by serological tests, as well as post-mortem examination of affected animals. A genome-wide scan was then performed on the individual marker genotypes, in an attempt to identify genomic regions associated with MAP infection disease. Nine significant markers were highlighted and they were located within, or nearby, annotated genes. Two genes found in this study encode are linked to protein kinases that are among the most important enzymes involved in the immune response to Johne’s disease, and four genes are involved in the functions of the Golgi complex.     

Rhyzopertha dominica (F., 1792) (Coleoptera: Bostrichidae): a stored grain pest on olive trees in Sicily

In October 2014 samples were collected from olive trees growing in South‐Eastern Sicily (Italy) that were showing wilting symptoms. In the field, clear signs of flagging appeared on young twigs and lateral shoots, and many leaves had fallen to the ground. Initial laboratory examinations of attacked twigs revealed the presence of a large number of adults of Rhyzopertha dominica (F., 1792). Sampling was repeated during the following 2 years, and while an outbreak of the pest did not occur, continuous damage was observed. In 2016, exclusively using pheromone‐baited traps in olive trees, a large number of adults were captured. The main local climatic parameters were monitored from 2014 to 2016 and compared with the population levels. The incidence of damage and survival of R. dominica in an environment different from its usual one (stored grain) deserves attention, especially considering that climate change could greatly affect pest demographics and put olive trees in this area and further afield at risk of increased damage.     

<Emphasis Type="Italic">Urochloa ruziziensis</Emphasis> cover crop increases the cycling of soil inositol phosphates

Ruzigrass (Urochloa ruziziensis) is a cover crop that is commonly used in Brazil and exudes high concentrations of organic acids from its roots, and is therefore expected to mobilize soil organic P such as inositol phosphates. However, it is not known if this can occur only under P deficient conditions. Specifically, we aimed to test the hypothesis that the degradation of inositol phosphates is increased by growing ruzigrass at two different P levels. To investigate this, we studied soil organic P in a 9-year-old field experiment, with treatments consisting of ruzigrass or fallow during the soybean (Glycine max) off-season, with or without P addition. Organic P was extracted in NaOH-EDTA, followed by colorimetric quantification of organic P hydrolysable by phytase, and myo-inositol hexakisphosphate by hypobromite oxidation and HPLC separation. Ruzigrass dry matter yield increased by about 80% with P application. Ruzigrass reduced the concentration of phytase labile P and myo-inositol hexakisphosphate, but only in soil receiving P. A corresponding increase in unidentified inositol phosphates, presumably representing lower-order esters, was also observed after ruzigrass in soil with P application. We deduce that the degradation of inositol phosphates under ruzigrass with P application is due to greater ruzigrass productivity in the more fertile treatment, increasing the release of root exudates that solubilize inositol phosphates and promote their decomposition by phytase. We conclude that ruzigrass cover cropping can promote the cycling of recalcitrant soil organic P, but only when fertility is raised to a sufficient level to ensure a productive crop.