Pathogenic diversity of Phytophthora sojae pathotypes from Brazil

Phytophthora root and stem rot has developed in commercial soybean fields since 2006 in Brazil, and cultivars with resistance to this disease have not been targeted for this region. Thus, the Phytophthora sojae pathotypes are expected to have virulence to few if any of the Rps genes. The objectives of this study were to characterize the pathotype diversity of P. sojae in Brazil, determine the distribution of the pathogen and predict which Rps genes will be effective and should be used in breeding programs. Isolates were collected in six states (Rio Grande do Sul, Santa Catarina, Paraná, Mato Grosso do Sul, Minas Gerais, and Goiás). The virulence formulae were based on the response of a differential set with 14 Rps genes (1a, 1b, 1c, 1d, 1k, 2, 3a, 3b, 3c, 4, 5, 6, 7, and 8). None of the 17 pathotypes found was reported previously. The most common virulence formulas were: 1d, 2, 3c, 4, 5, 6, 7 (octal code 05471, representing 24 % of the occurrences); 1d, 2, 3b, 3c, 4, 5, 6, 7 (05671, 13 %); 1b, 1d, 2, 3a, 3c, 4, 5, 6, 7 (25571, 8 %); and 1d, 3a, 5, 7, 8 (01123, 8 %). Percentages of isolates with a susceptible interaction with each Rps gene was Rps1a (3 %), Rps1b (11 %), Rps1c (3 %), Rps1d (100 %), Rps1k (3 %), Rps2 (86 %), Rps3a (32 %), Rps3b (19 %), Rps3c (73 %), Rps4 (70 %), Rps5 (89 %), Rps6 (59 %), Rps7 (100 %), and Rps8 (22 %). There was apparently no relationship between pathotypes and origin. Stacking resistance genes Rps1a, Rps1b, Rps1c, and Rps1k with Rps3b or Rps8 would be highly effective for soybean cultivars targeted for Brazil.     

Urolithins Are the Main Urinary Microbial-Derived Phenolic Metabolites Discriminating a Moderate Consumption of Nuts in Free-Living Subjects with Diagnosed Metabolic Syndrome

Walnuts ( Juglans regia L.), hazelnuts ( Corylus avellana L.), and almonds ( Prunus dulcis Mill.) are rich sources of ellagitannins and proanthocyanidins. Gut microbiota plays a crucial role in modulating the bioavailability of these high molecular weight polyphenols. However, to date there are no studies evaluating the capacity to produce nut phenolic metabolites in subjects with metabolic syndrome (MetS), a pathology associated with an altered gut bacterial diversity. This study applied a LC-MS targeted approach to analyze the urinary excretion of nut phenolic metabolites in MetS subjects following 12 weeks of nut consumption, compared to sex- and age-matched individuals given a nut-free control diet. Metabolites were targeted in both hydrolyzed and nonhydrolyzed urine by LC-PDA-QqQ-MS/MS analysis, and identification of metabolites lacking available standards was confirmed by LC-ESI-ITD-FT-MS. Ellagitannin-derived urolithins A and B significantly increased after the nut-enriched-diet, urolithins C and D were also detected, and a complex combination of urolithin-conjugated forms was observed in nonhydrolyzed urine, confirming an extensive phase II metabolism after absorption. In contrast, no significant increases in proanthocyanidin microbial metabolites were observed in urine following nut consumption. Because the intestinal microbiota of the subjects in this study could catabolize ellagitannins into a wide range of urolithins, further research is strongly warranted on the in vivo potential of these microbial metabolites in reducing cardiometabolic risk.     

Polyphenols and health: current state and progress

During the 5th International Conference on Polyphenols and Health that was held in Sitges (Spain) in October 2011, the latest advances in this area of active research were presented. Sessions on polyphenol effects on cardiovascular disease, polyphenols as ingredients of functional foods, the role of polyphenols in preventing obesity and diabetes, the interaction of polyphenols with gut microbiota, bioavailability and metabolism of polyphenols in humans, the mechanisms of action of these metabolites in different models, new methodologies for the study of the role of polyphenols in health, polyphenols and cancer, recent developments in phenolic compounds and neuroscience, and polyphenols in epidemiology and public health were organized. This highlight issue presents a selection of papers from invited speakers, oral presentations, and poster prize winners. The perspectives for this exciting area of very active research were also discussed at the meeting and are summarized in this introductory paper.     

Isoelectric focusing of β-glucosidase humic-bound activity in semi-arid Mediterranean soils under management practices

Changes in β-glucosidase enzyme–humic complexes and conventional parameters (pH, total organic C, total N, water-soluble C, and bulk density) were studied in an almond-cropped soil prone to erosion under a rehabilitation practice. The experimental plan included three soil slopes (0%, 2%, and 6%) and two type of fertilization (organic and mineral), with sampling of rhizosphere and inter-row soils. The enzyme humic complexes were extracted by pyrophosphate, purified by ultrafiltration of the organic extracts on molecular mass exclusion membranes (mol wt > 10⁴) and fractionated by isoelectric focusing technique (IEF). The IEF on polyacrylamide rod gels with a restricted pH gradient ranging between 6.0 and 4.0 gave five humic bands on the basis of the little differences of their electric charges (pI). Under both organic and mineral fertilization, β-glucosidase activity bound to the fractionated humic substances, especially in the pH range 4.5–4.2 of the rhizosphere soil, was higher than that of the inter-row soil. This also occurred in 6% slope where the enzyme activity was lower than in soil with lower slopes. The higher number of the β-glucosidase active humic bands in rhizosphere than inter-row soil, particularly for the 0% slope, may be due to the presence of humic molecules capable of preserving the enzyme molecules in the active form, other than to the higher microbial activity synthesizing and releasing the tested enzymes.     

Metal fluxes at the sediment–water interface in rivers in the Turvo/Grande drainage basin, S?o Paulo State, Brazil

Purpose

The Turvo/Grande drainage basin (TGDB), located in the northwestern region of S?o Paulo state, covers an area of 15,983?km². The region is typically regarded as agricultural by the S?o Paulo State Environmental Agency, but the industrial area is expanding, and some studies have shown that metal concentrations in water can be higher than the values regulated by Brazilian law. Therefore, the aim of this study was to assess the role of sediments as a source or a sink of metals for drainage basin management.

Materials and methods

Interstitial water from different sediment depths (0?C42?cm) and the sediment?Cwater interface and sediment core samples were collected in February and July 2010 from the Preto, Turvo, and Grande rivers. Quantification of Cr, Cu, Fe, Mn, Ni, and Pb in these samples was performed by graphite furnace or flame atomic absorption spectrometry. Metal diffusive flux estimation from sediment into the overlying water was calculated by Fick??s First Law of Diffusion.

Results and discussion

The fluxes of all metals for the three rivers were positive, indicating diffusion into the overlying water. Ni and Pb showed the lowest diffusive fluxes, which ranged from 2.4 to 3,978???g?m^?2?day^?1 for Ni and from ?0.1 to 1,597???g?m^?2?day^?1 for Pb. In turn, Cu and Cr were subject to the largest transfer to water, especially in the dry season (Cr, 4.5?C7,673???g?m^?2?day^?1; Cu, 1.3?C14,145???g?m^?2?day^?1). The Preto River (urban area) showed smaller fluxes than the Grande River (agricultural area), and the values of the latter were higher than those found in other impacted areas of the world.

Conclusions

The diffusive fluxes indicate that sediments from the TGDB act as a source of metals for the water column, with increased export of metals, particularly Cr and Ni, from the sediment into the overlying water during the dry season.     

EFFECT OF SEVOFLURANE ANESTHESIA AND BLOOD DONATION ON THE SONOGRAPHIC APPEARANCE OF THE SPLEEN IN 60 HEALTHY CATS

The aims of this prospective study were to describe the normal sonographic size and appearance of the feline spleen, and to test effects of sevoflurane anesthesia and blood collection on sonographic characteristics. Sonographic evaluations were performed in 60 healthy blood donor cats prior to anesthesia and blood collection (baseline), after anesthesia induction, after completion of blood collection while under anesthesia, and 7–10 days after blood collection. Transverse sonographic images of the splenic mid‐body were made. Splenic height, width, cross‐sectional area, echogenicity relative to the left renal cortex, and echotexture were recorded. Height, width, and cross‐sectional area were compared across time periods using linear‐mixed effects models. Holm's method was applied to adjust for multiplicity and control the overall Type 1 error rate at α = 0.05. Normal transverse sonographic splenic measurements (mean ± standard deviation) at baseline were: height 8.2 ± 1.4 mm; width 26.7 ± 4.4 mm; and CSA 1.6 ± 0.5 cm². While there were statistically significant differences in splenic height, width, and cross‐sectional area after sevoflurane anesthesia and/or blood collection, differences were of low magnitude. The spleen was diffusely homogenous and subjectively isoechoic or mildly hyperechoic relative to the left renal cortex. In 77% of castrated male cats, the spleen was isoechoic to the left renal cortex. We conclude that sevoflurane anesthesia and blood donation do not subjectively alter splenic dimensions, echogenicity and echotexture in normal cats.     

Comparative endocrinological responses to short transportation of Equidae (Equus asinus and Equus caballus)

In order to evaluate the effects of short transportation on β‐endorphin, adrenocorticotropic hormone (ACTH) and cortisol changes, 12 healthy stallions of Equidae (Equus asinus and Equus caballus) were studied before and after transportation of 50 km. Blood samples were collected 1 week before transportation in basal conditions, immediately before loading and after transportation and unloading, on their arrival at the breeding station. Compared to basal and before values, donkeys showed an increase in circulating ACTH (P < 0.001) and cortisol (P < 0.0005) levels after transportation and higher ACTH (P < 0.01) levels than horses after transportation. A positive and significant correlation (r = 0.885; P < 0.01) between ACTH and cortisol levels after transportation was found. No significant differences were observed for β‐endorphin levels. Compared to basal and before values, horses showed higher cortisol (P < 0.005) levels after transportation and no significant differences were observed for ACTH and β‐endorphin levels in donkeys. Horses facing forward (direction of travel) showed higher (P < 0.01) β‐endorphin levels after transportation than donkeys; horses facing backward (the opposite direction of travel) showed lower (P < 0.05) ACTH levels after transportation. The results indicate that short transportation induces a preferential activation of the hypothalamus‐pituitary‐axis (HPA), with significant release of ACTH and cortisol in donkeys and only of cortisol in horses, suggesting that transportation for donkeys may be more stressful than horses.     

Efficacy of a BVDV subunit vaccine produced in alfalfa transgenic plants

Bovine viral diarrhea virus (BVDV) is considered an important cause of economic loss within bovine herds worldwide. In Argentina, only the use of inactivated vaccines is allowed, however, the efficacy of inactivated BVDV vaccines is variable due to its low immunogenicity. The use of recombinant subunit vaccines has been proposed as an alternative to overcome this difficulty. Different studies on protection against BVDV infection have focused the E2 protein, supporting its putative use in subunit vaccines. Utilization of transgenic plants expressing recombinant antigens for the formulation of experimental vaccines represents an innovative and cost effective alternative to the classical fermentation systems.The aim of this work was to develop transgenic alfalfa plants (Medicago sativa, L.) expressing a truncated version of the structural protein E2 from BVDV fused to a molecule named APCH, that target to antigen presenting cells (APCH-tE2). The concentration of recombinant APCH-tE2 in alfalfa leaves was 1 μg/g at fresh weight and its expression remained stable after vegetative propagation. A methodology based an aqueous two phases system was standardized for concentration and partial purification of APCH-tE2 from alfalfa. Guinea pigs parentally immunized with leaf extracts developed high titers of neutralizing antibodies. In bovine, the APCH-tE2 subunit vaccine was able to induce BVDV-specific neutralizing antibodies. After challenge, bovines inoculated with 3 μg of APCH-tE2 produced in alfalfa transgenic plants showed complete virological protection.