alpha-Hydroxynitrile lyase protein from Xylella fastidiosa: Cloning, expression, and characterization

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Análise fenotípica e molecular da produção de biofilmes por estirpes de staphylococcus aureus isoladas de casos de mastite subclínica bovina

Biofilm formation is considered an advantage for Staphylococcus aureus mastitis isolates, facilitating bacterial persistence in the udder. It requires attachment to mammary epithelium, proliferation and accumulation of cells in multilayers and enclosing in a polymeric matrix known as exopolysaccharide. The objective of this study was to evaluate the ability of Staphylococcus aureus isolated from bovine subclinical mastitis for formation of biofilms. A total of 94 Staphylococcus aureus strains obtained from milk samples of cows suffering from subclinical mastitis in dairy herds on two properties in the state of São Paulo were evaluated. These strains were characterized by in vitro biofilm formation, and by the presence of icaA and icaD genes which are responsible for intercellular adhesion. The results revealed that 98.9% of the isolates produced biofilm in vitro by adherence in sterile 96-well "U" bottom polystyrene tissue culture plates; 95.7% of the isolates possessed the icaA and icaD genes. These bacterial isolates biofilm producers may impair eradication of chronic mastitis, rendering antibiotherapy less effective. The detection of biofilm forming ability in mastitis isolates may provide useful information for more adequate therapeutic regimen and for preventive actions in the control of those bacterial isolates in bovine herds.

Reação do solo e disponibilidade de micronutrientes, em solo de textura média, tratado com lodo de esgoto e cultivado com milho

The evaluation of micronutrient availability in agricultural soil treated with sewage sludge was developed in Field with maize cultivation on a Dystrophic Red Latosol. The experiment used a randomized complete block design, with the treatments in a split-plot scheme, in time, with six doses of sewage sludge (main-plot treatment) in relation to the five sampling periods for the soil (subplot treatment), with four replications. The sewage sludge was applied in the superficial layer (0-20 cm). The micronutrients Fe, Cu, Mn and Zn were extracted from the soils using a DTPA (diethylenetriaminepentaacetic acid) solution. The same micronutrients were analyzed in the plant tissue after the nitric-perchloric decomposition of the maize samples by spectrometric atomic absorption. The results showed an increasing amount of Fe, Mn, Zn, Cu, Cation Exchange Capacity (CEC) and a decreasing pH in the soil. The increasing doses of sewage sludge increased the Fe, Mn, Cu and Zn availability in the soil solution, and the uptake amount by maize plants were always greater than the control values.

Bull semen varibles after experimental exposure with Bovine Herpesvirus type 5

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Desenvolvimento in vitro de copo-de-leite: efeito das concentrações de sacarose e de ácido giberélico

Calla lily (Zantedeschia aethiopica) is appreciated as cut flower and for the composition of gardens. However, many pathogens affect this species. By the traditional method of propagation, some units of new seedlings can only be produced annually. Tissue culture allows fast large-scale clonal propagation and provides healthy uniform plants. During the in vitro process, type and concentration of growth regulator could affect the growth of seedlings. Thus, the aim of this work was to determine sucrose and GA(3) concentrations to increase the efficiency of the in vitro multiplication of calla lily. After 60 days, the length of the above ground part and the roots, the number of sprouts, roots and leaves, above ground part and root fresh weight of seedlings were evaluated. The experimental design was entirely randomized with four replications. It was necessary the addition of 60.5 g L-1 sucrose associated to 5 mg L-1 GA(3) to obtain hight sprouts number. For higher length of the above ground part the addition of 45.3 g L-1 sucrose and 10 mg L-1 GA(3) was enough. Better results in the root length and number of roots were observed only in the sucrose presence, in concentrations in the range of 51.13 - 56.5 g L-1.

Histoplasmose do esôfago: Relato de caso

The authors report a case of a patient with complaint of progressive disphagia. Stenoses of lower third of esophagus was revealed by radiological and endoscopic examinations. Fungi were showed in biopsy of lesion, with demonstration of Histoplasm capsulate by tissue culture. Endoscopic dilatation was performed because especific medical treatment failed but esophageal rupture was observed. Partial esophagectomy was performed with symptoms remission.

Vegetative propagation of Mikania glomerata: Micro-propagation and cuttings

The scope of this work was to compare two systems for vegetative propagation: conventional one (from cut stems) and in vitro micropropagation from axillary buds. Nodal segments (1 cm) of Mikania glomerata were used as explants. The experiments were evaluated in relation to number of shoots; % of rooting; number of roots and total fresh weight. Multiple shoots developed in MS containing 0.5 mg/L BAP. Rooting was induced in the presence of 1.0 mg/L IBA. Stems with five buds and one pair of leaves were the most appropriate for the production of cuttings. The time necessary for developing a protocol for the production of M. glomerata micropropagated plantlets was 6 months, whereas only half time was required to produce plantlets from stem cuttings. The greatest problem met during micropropagation was the culture contamination by endophytic bacteria and fungi.

Produção de composto orgânico a partir de residuos de plantas medicinais

Extraction of active principles from medicinal plants generates wastes usually discharged. This process does not extract minerals from plant tissues, which may be considered of great value in organic composting. Wastes of 81 medicinal plant species processed at Centroflora/Anidro industry were analyzed for macro and micronutrients, and according to the results, three different ways to elaborate the compost were proposed: a) wastes of medicinal plants with high content of a specific mineral, aiming to fertilize soils where would be cultivated species that need high quantities of that mineral; b) wastes with mineral contents in order to support specific failures on mineral content of a soil; c) wastes of the eleven richest species in each nutrient that should elevate all mineral contents in soil.

Efeito do alumínio no crescimento de brotações de Eucalyptus grandis x E. urophylla cultivadas in vitro

The aim of this work was to evaluate the effects of aluminum on the growth of Eucalyptus shoots cultivated in vitro through nutrient and total soluble protein content. The trial had a totally randomized design with four treatments and four replicates. The treatments were: 0.0; 0.25; 0.5 and 1.0 mM of AlCl 3.6H 2O. Shoots without roots of a Eucalyptus grandis x E urophylla clone were used for the in vitro culture. Evaluations were made on the 4th, 8th, 12th, 16th, 20th, 24th and 28th day of culture. The Al addition to the culture media reduced mainly Ca, P and K availability and absorptions by the shoots. The cellular metabolism was affected, conducted to morphological alterations in shoots (browning, mass calluses formation and shoots not friable), dry matter increased and a decreased in total protein soluble.

Effect of capacitation of stallion sperm with polyvinylalcohol or bovine serum albumin on penetration of bovine zona-free or partially zona-removed equine oocytes

Experiments were conducted to study effects of macromolecules on stallion sperm capacitation and fertilization as determined by penetration of bovine zona-free and equine partially zona-removed oocytes. Stallion sperm were capacitated in TYH medium (modified Krebs-Ringer bicarbonate) supplemented with either 1 mg/mL of polyvinylalcohol (PVA) or 4 mg/ mL of BSA. Capacitation was induced with 8 bromoadenosine cyclic monophosphate (8BrcAMP; 0.5 mM) alone or in combination with 0.1 μM of ionomycin. Intraspecies gametes were co-incubated in TYH/PVA or TYH/ BSA for 18 to 20 h. For zona-free bovine oocytes, penetration rate (35%) with the combination of 8BrcAMP and ionomycin in PVA-containing medium was higher (P < 0.05) than any treatment in BSA-containing medium (5 to 6%). A similar study was conducted using equine oocytes with partially removed zonae. Sperm capacitated and used for in vitro fertilization (IVF) in PVA-containing medium had higher penetration rates (P < 0.01) than sperm in BSA-containing medium (54 vs. 11%). The effect of equine preovulatory follicular fluid on bovine oocyte penetration was assessed. Bovine oocytes were matured in tissue culture medium-199 with 0, 20, 50, or 100% equine preovulatory follicular fluid, and 1 IU/mL of equine chorionic gonadotropin. Stallion sperm were treated with 8BrcAMP + ionomycin in PVA- or BSA-containing media. The penetration rates of bovine zona-free oocytes by stallion sperm were again higher with PVA (47%) than BSA (18%; P < 0.01). Penetration rates of oocytes matured in 100% follicular fluid were higher (P < 0.05) than for oocytes matured with 0% follicular fluid. The effects of equine follicular fluid and PVA/BSA during sperm capacitation on standard bovine IVF were examined. Culture of bovine oocytes with equine follicular fluid did not affect oocyte maturation or penetration rates after IVF. Bovine sperm capacitated with heparin in PVA-containing medium yielded lower (P < 0.05) fertilization rates than those capacitated in BSA-containing medium when incubated with both zona-intact and zona-free bovine oocytes. In summary, PVA was superior to BSA for ionophore-induced capacitation of equine sperm for penetration of zona-free bovine oocytes or partially zona-removed equine oocytes, but not for standard bovine IVF with bovine sperm. Zona-free bovine oocytes may be useful for assaying in vitro capacitation and fertilization of stallion sperm. © 2003 American Society of Animal Science. All rights reserved.

Putative pyrophosphate phosphofructose 1-kinase genes identified in sugarcane may be getting energy from pyrophosphate

Pyrophosphate-dependent phosphofructokinase (PPi-PFK) has been detected in several types of plant cells, but the gene has not been reported in sugar cane. Using Citrux paradixi PPi-PFK gene (AF095520 and AF095521) sequences to search the sugar cane EST database, we have identified both the α and β subunits of this enzyme. The deduced amino acid sequences showed 76 and 80% similarity with the corresponding α and β subunits of C. paradisi. A high degree of similarity was also observed among the PFK β subunits when the alignment of the sugar cane sequences was compared to those of Ricinus communis and Solanum tuberosum, it appears that α and β are two distinct subunits; they were found at different concentrations in several sugar cane tissues. It remains to be determined if the different gene expression levels have some physiological importance and how they affect sucrose synthesis, export, and storage in vacuoles. A comparison between the amino acid sequences of β PFKs from a variety of organisms allowed us to identify the two critical Asp residues typical of this enzyme's activity site and the other binding sites; these residues are tightly conserved in all members of this protein family. Apparently, there are catalytic residues on the β subunit of the pyrophosphate-dependent enzyme.

Bud source, asepsis and benzylaminopurine (BAP) effect on yacon (Polymnia sonchifolia) micropropagation

The yacon (Polymnia sonchifolia) is used largely for the high fructan content of its tubers; consequently, it is a good alternative for diabetics. One of the more important restricting factors of the commercial production of yacon is its susceptibility to nematode attack. This, as well as germplasm bank maintenance, justifies the importance of in vitro propagation of this species. In this way, our work aimed to verify the best asepsis method for yacon for the in vitro establishment from the rhizophore and the axillary buds of the aerial parts, and the effect of benzylaminopurine (BAP) addition to the culture medium. The number of contaminated cultures, the occurrence of phenolic oxidation and the occurrence of a vitreous aspect, showed differences with bud source, immersion time for asepsis, and BAP use. The results contribute to establishing a yacon micro propagation procedure.

Effect of benzylaminopurine (BAP) on clonal propagation rate of Curcuma longa L.

Curcuma longa L. is used in many countries for its flavor, and medicinal and cosmetic attributes, as well as for its peculiar starch characteristics. These factors have driven an interest in the in vitro propagation of this species, looking for germplasm bank maintenance, production of disease free plants, genetic variability induction from callus, and as a tool for starch research. However, there are few reports concerning the micropropagation of Curcuma longa. The in vitro propagation rate of this species, cultured under two benzylaminopurine (BAP) concentrations, was the aim of this research.

In silico evaluation of the Eucalyptus transcriptome

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Enxertia e água de irrigação carbonatada no transporte de 15N e na produção do tomateiro

The effects of CO2 application through irrigation water, and of grafting in transport of 15N and in the tomato production, were studied. These treatments were arranged in a 2 x 2 factorial scheme (with and without CO2 in irrigation water and grafted and non-grafted tomato), in a completely randomized design, with four replications. The injection of CO2 into the water began at 34 days after transplant of seedlings (DAT) and continued for all irrigations. The application of the sulfate of ammonium with abundance in atoms of 15N of 3.13% in plants destined to analysis was done at 45 DAT when the plants were in the middle of fructification. After 14 days of fertilizer (15N) application the plants were harvested, washed, dried and sent for analysis of 15N in plant tissue. The results demonstrated that CO2 and the grafting did not alter the transport of 15N in the plant. The production of commercial fruits was larger when CO2 was applied in water.