Hentriacontane: a leaf hydrocarbon from Syzygium jambos with stimulatory effects on the germination of urediniospores of Puccinia psidii

A crude Sohxlet extract from leaves of Syzygium jambos was sequentially fractionated using a silica gel flash column. A bioassay based on the numbers of urediniospores of Puccinia psidii that germinated in 2% water agar detected an active stimulant of germination when the fraction eluted with 100% n-hexane was used. The active fraction induced up to 88% increase in germination when added to a spore suspension in mineral oil. The active fraction was characterized as a hydrocarbon by ¹H nuclear magnetic resonance, 13C nuclear magnetic resonance, and infrared analysis. Gas chromatography-mass spectrometry analysis indicated that the fraction was a long-chain 436 MW hydrocarbon with corresponding to C31H64, namely hentriacontane. This is the first time such a compound proved to be involved with stimulation of fungal spore germination. These results may contribute to better understanding the infection process of rusts.

RT-PCR-ELISA for detection of Apple stem grooving virus in apple trees

A method to detect Apple stem grooving virus (ASGV) based on reverse transcription polymerase chain reaction (RT-PCR) was developed using primers ASGV4F-ASGV4R targeting the viral replicase gene, followed by a sandwich hybridisation, in microtiter plates, for colorimetric detection of the PCR products. The RT-PCR was performed with the Titan™ RT-PCR system, using AMV and diluted crude extracts of apple (Malus domestica) leaf or bark for the first strand synthesis and a mixture of Taq and PWO DNA polymerase for the PCR step. The RT-PCR products is hybridised with both a biotin-labelled capture probe linked to a streptavidin-coated microtiter plate and a digoxigenin (DIG)-labelled detection probe. The complex was detected with an anti-DIG conjugate labelled with alkaline phosphatase. When purified ASGV was added to extracts of plant tissue, as little as 400 fg of the virus was detected with this method. The assay with ASGV4F-ASGV4R primers specifically detected the virus in ASGV-infected apple trees from different origins, whereas no signal was observed with amplification products obtained with primers targeting the coat protein region of the ASGV genome or with primers specific for Apple chlorotic leaf spot virus (ACLSV) and Apple stem pitting virus (ASPV). The technique combines the power of PCR to increase the number of copies of the targeted gene, the specificity of DNA hybridization, and the ease of colorimetric detection and sample handling in microplates.

Identification of the major fungitoxic component of cinnamon bark oil

The study was done to identify the most active fungitoxic component of cinnamon bark (Cinnamomum zeylanicum) oil that can be used as a marker for standardization of cinnamon extract or oil based natural preservative of stored seeds. Aspergillus flavus and A. ruber were used as test fungi. The hexane extracted crude oil and the hydro-distilled essential oil from cinnamon bark had complete growth inhibition concentration (CGIC) of 300 and 100 µl/l, respectively. Both oils produced three fractions on preparative thin layer silica-gel chromatography plates. The fraction-2 of either oil was the largest and most active, with CGIC of 200 µl/l, but the fungitoxicity was also retained in the other two fractions. The fraction-1 and 3 of the crude oil reduced growth of both the fungal species by 65%, and those of distilled oil by 45% at 200 µl/l. The CGIC of these fractions from both the sources was above 500 µl/l. The gas chromatography and mass spectrometry (GC-MS) of the fraction-2 of the hexane extract revealed that it contained 61% cinnamaldehyde, 29% cinnamic acid, and two minor unidentified compounds in the proportion of 4% and 6%. The GC-MS of the fraction-2 of the distilled oil revealed that it contained 99.1% cinnamaldehyde and 0.9% of an unidentified compound. The CGIC of synthetic cinnamaldehyde was 300 µl/l and that of cinnamic acid above 500 µl/l. The 1:1 mixture of cinnamaldehyde and cinnamic acid had CGIC of 500 µl/l. The data revealed that cinnamaldehyde was the major fungitoxic component of hexane extract and the distilled essential oil of cinnamon bark, while other components have additive or synergistic effects on total fungitoxicity. It is suggested that the natural seed preservative based on cinnamon oil can be standardized against cinnamaldehyde.

Allelopathic potential and systematic evaluation of secondary compounds in extracts from roots of Canavalia ensiformis by capillary electrophoresis

Organic extracts were obtained from roots of Canavalia ensiformis and evaluated for allelopathic potential on the germination of the weed seeds: Mimosa pudica, Cassia tora and Cassia occidentalis showing a strong allelopathic potential. After that, a systematic study of these crude extracts was made using specific protocols developed in capillary electrophoresis (CE) in order to determine some classes of secondary metabolites. Capillary electrophoresis protocols were highly specific, which makes it possible to identify 5 classes of compounds using the same crude extract samples and analyze them fartly. Some of the compounds identified show activity in the inhibition of seeds germination.

Antimicrobial activity of extracellular metabolites from antagonistic bacteria isolated from potato (Solanum phureja) crops

Microorganisms for biological control are capable of producing active compounds that inhibit the development of phytopathogens, constituting a promising tool toob tain active principles that could replace synthetic pesticides. This study evaluatedtheability of severalpotentialbiocontrol microorganismsto produce active extracellular metabolites. In vitro antagonistic capability of 50 bacterial isolates from rhizospheric soils of "criolla" potato (Solanum phureja) was tested through dual culture in this plant with different plant pathogenic fungi and bacteria. Isolates that showed significantly higher antagonistic activity were fermented in liquid media and crude extracts from the supernatants had their biological activities assessed by optical density techniques. Inhibitory effecton tested pathogens was observed for concentrations between 0.5% and 1% of crude extracts. There was a correlation between the antimicrobial activity of extracts and the use of nutrient-rich media in bacteria fermentation. Using a bioguided method, a peptidic compound, active against Fusarium oxysporum, was obtained from the 7ANT04 strain (Pyrobaculum sp.). Analysis by nuclear magnetic resonance and liquid chromatography coupled to mass detector evidenced an 11-amino acid compound. Bioinformatic software using raw mass data confirmed the presence of a cyclic peptide conformed by 11 mostly non-standard amino acids.

Prevalence of Anxiety and Depression among Medical Students

ABSTRACT Background Medical students are a vulnerable population to develop depression and anxiety disorders. Objective To estimate the prevalence and associated factors of anxiety and depression among medical students. Methods A cross-sectional study with a random sample (n = 346) of medical students at a Brazilian university was performed. The Beck Depression Anxiety Inventory was used to measure anxiety and depression levels. Crude and adjusted analyses were performed using Poisson regression. Results The prevalence of anxiety was 35.5% and depression was 32.8%. The prevalence of anxiety and depression was 14% higher and 16% higher among women (p = 0.025 and p = 0.006, respectively). Students whose parents were not physicians reported 23% higher prevalence of anxiety (p = 0.006), and those who had physician parents reported 29% higher prevalence of depression (p = 0.034). Those who always or often felt pushed by their parents showed 22% higher prevalence of anxiety (p =.006) and 19% higher depression (p = 0.016). Students who had concerns over the future had 15% higher prevalence of depression (p = 0.017). Conclusion The prevalence of anxiety and depressive symptoms was higher than the average found in the general population.

Effect of light intensity and growth substratum on plant development and production of secondary metabolites in Cordia curassavica (Jacq.) Roem. & Schult

Cordia curassavica (Jacq.) Roem. & Schult. (Boraginaceae), also referred to as Cordia verbenacea DC, has been traditionally used for medicinal purposes. This study was driven to verify the behavior of the species in similar conditions to its natural environment, such as high light intensity and sandbank soil, and in conditions of low light intensity and fertilized substratum (dystroferric red nitosoil plus earthworm humus). The growth of the plant, the income of leaf crude extracts and, in the alcoholic extract, the number of substances found in thin layer cromatography and the toxicity of the substratum was observed. The results indicated that the growth of the root biomass, stem and leaves in discharge or lower light intensity was similar, but smaller in sandbank soil than in fertilized soil. The relative income of extracts in ether of petroleum and alcohol was larger in high light intensity and fertilized substratum. The light intensity and the substratum type didn't affect the number of substances detected in the alcoholic extract or the toxicity of this extract. Stains corresponding to the rosmarinic acid were only evidenced in some samples of the alcoholic extract, not allowing the verification of the effect of the treatments about its production.

Recycling of nutrients with application of organic waste in degraded pasture

The utilization of organic wastes represents an alternative to recover degraded pasture. The experiment aimed to assess the changes caused by the provision of different organic waste (poultry litter, turkey litter and pig manure) in a medium-textured Oxisol in Brazilian Savanna under degraded pasture. It was applied different doses of waste compared to the use of mineral fertilizers and organic mineral and evaluated the effect on soil parameters (pH, organic matter, phosphorus and potassium) and leaf of Brachiariadecumbens (crude protein, phosphorus and dry mass production). It was observed that application of organic waste did not increase the level of soil organic matter and pH in the surface layer, and the application of turkey litter caused acidification at depths of 0.20-0.40 m and 0.40-0.60 m. There was an increase in P and K in the soil with the application of poultry litter and swine manure. All organic wastes increased the productivity of dry matter and crude protein and phosphorus. The recycling of nutrients via the application of organic waste allows efficiency of most parameters similar to those observed with the use of mineral sources, contributing to improving the nutritional status of soil-plantsystem.

Effect of the application of different water depths and nitrogen and potassium doses on quality of Tanzania grass

The objective of this study was to evaluate the effects of the application of different water depths and nitrogen and potassium doses in the quality of Tanzania grass, in the southern of the state of Tocantins. The experiment was conducted on strips of traditional sprinklers, and used, as treatments, a mixture of fertilizer combinations of N and K2O always in the ratio of 1 N:0.8 K2O. This study determined throughout the experiment: plant height (PH), the crude protein (CP) and neutral detergent fiber (NDF). The highest plant height obtained was 132.4 cm, with a fertilizer dose of 691.71 kg ha-1 in the proportion of N:0.8 K2O, in other words, 384.28 kg ha-1 of N and 307.43 kg ha-1 of K2O, and water depth of 80% of the ETc. The highest crude protein content was 12.2%, with the fertilizer dose application of 700 kg ha-1 yr-1 in the proportion of 1 N to 0.8 of K2O, in other words, 388.89 kg ha-1 of N and 311.11 kg ha-1 of K2O and absence of irrigation. The lowest level of neutral detergent fiber was 60.7% with the application of the smallest dose of fertilizer and highest water depth. It was concluded in this study that there was an increase in plant height by increasing the fertilizer dose and water depth. The crude protein content increased 5.4% in the dry season, by increasing the fertilizer dose and water depth. In the dry season, there was an increase of NDF content by 4.5% by increasing the application of fertilizer and water depth.

Control of ammonia emissions in naturally ventilated dairy cattle facilities in Portugal

We conducted a study of the processes associated to NH3 emission in naturally ventilated dairy cattle facilities, having described factors that regulate NH3 emission, as well as methodologies for measuring these emissions at these facilities. Appropriate techniques to mitigate NH3 emission in facilities located in regions with warm climates were also identified. The most effective mitigation techniques with simple implementation include strategies associated to: (i) installation design and flooring, which lead to reduced emissions, (ii) excreta pre-excretion, namely the use of diets with optimized crude protein content and increased milk production at farm level; and (iii) excreta post-excretion, particularly by changing the conditions of environmental monitoring within the premises, practice introduction or additive application in the management of excreta deposited on floors.

Serodiagnosis of Babesia equi in horses submitted to exercise stress

A complement fixation test (CFT), performed in microtitre plates, based upon the use of crude antigenic preparation of Babesia equi was adapted for the detection of antibodies in serum of infected horses. The indirect fluorescent antibody test (IFAT) and enzyme-linked immunosorbent assay (ELISA) were also used for the immunodiagnosis of B. equi. Serum samples from 15 apparently healthy horses, previously conditioned to a high-speed equine treadmill, were taken before and after exercise. All the samples analyzed were positive for B. equi infection. There were no significant differences (P<0.01) between these 3 tests, or the condition of rest or stress. The combined use of CFT and IFAT or ELISA should be recommended in order to enable veterinary services to more efficiently prevent introduction of infected horses into disease-free areas.

Efficacy of bacterin-, outer membrane protein- and fimbriae extract-based vaccines for the control of Salmonella Enteritidis experimental infection in chickens

The efficacy of three vaccines was evaluated in chickens for the control of experimental infection with Salmonella Enteritidis (SE) phage type 4. The vaccines were produced with bacterin, outer membrane proteins (OMP) and fimbriae crude extract (FE). The chickens were vaccinated intramuscularly with two doses of each vaccine at 12 and 15 weeks of age. The chickens were then orally challenged with 10(9) CFU/chicken Salmonella Enteritidis phage type 4 at 18 weeks of age. Fecal swabs were performed for the recovery of shedding SE, and SE was recovered from the liver and spleen. Additionally, antibody titers were measured in the serum by micro-agglutination test. The results indicated that the vaccine produced with bacterin yielded better results and resulted in reduction of fecal shedding and organ invasion by SE after oral challenge, although no vaccine was 100% effective for the control of SE experimental infection.

Comparison of methods for determination of nutrient digestibility of a dry kibble diet for ocelots

In this experiment, methods of total fecal collection (TFC) and internal markers (acid-insoluble ash - AIA, crude fiber - CF, and acid-detergent fiber - ADF) were compared for determination of the coefficients of apparent digestibility (CAD) for dry matter (DM), crude protein (CP), ether extract (EE), nitrogen-free extracts (NFE), and gross energy (GE) of commercial feline dry kibble for ocelots (Leopardus pardalis). Six adult animals, weighing 12.45±1.37 kg, gradually received experimental kibble in their usual diet until the beginning of the experiment and were submitted to an adaptation period ten days prior to the collection period. CAD obtained by TFC, AIA, CF, and ADF were, respectively, 73.7, 76.83, 62.01, and 46.03% for dry matter; 81.9, 84.8, 75.8, and 63.8% for crude protein; 85, 86.7, 78.5, and 69.1% for ether extract; 78.52, 79.55, 69.11, and 53.04% for nitrogen-free extracts; and 80.5, 82.2, 71.4, and 58.4% for gross energy. The AIA method showed to be efficient in determining coefficients of apparent digestibility and may contribute to investigations on the digestibility of diets for wild felines. In comparison to the items of ocelot's usual diet, the kibble used in this paper provided an adequate nutritional supply with reduced daily costs per animal.

Effects of Corynespora cassiicola on Lantana camara

The present study combines the examination of toxins produced by C. cassiicola and the effects of the fungus colonization on L. camara. C. cassiicola was cultivated on solid media and the crude extracts CAE and CE were produced. Both extracts were submitted to a seed germination and growth assay utilizing Physalis ixocarpa, Trifolium alexandrinum, Lolium multiflorum and Amaranthus hypochodriacus. The effect of the extracts on the ATP-synthesis in isolated spinach chloroplasts was also tested. Bioassay guided chromatographic fractionation identified the most active extract (CAE). From this extract ergosta-4,6,8(14),22-tetraen-3-one (C1) and fatty acids were isolated. The C1 compound reduce ATP synthesis in isolated spinach chloroplasts. The interference of fatty acids with ATP synthesis and also with weed growth provides one explanation of the phytogrowth-inhibitory properties of such fungal extracts. Histological observations involving fungus-plant interaction were made on L. camara plants inoculated with C. cassiicola conidia suspension. After inoculations, fragments of the leaf blades were prepared for observation by light and scanning electron microscopy. Fungal colonization of Lantana camara was typical of a necrotroph and penetration initiated a hypersensitive response. L. camara reacted to the pathogen penetration through thickening of the epidermis walls, cytoplasm granulation and a cicatrisation tissue.

Effect of Serjania lethalis ethanolic extract on weed control

The objective of this study was to evaluate the effect of the ethanolic extract of Serjania lethalis leaves and stems on the diaspore germination and seedling growth of wild poinsettia (Euphorbia heterophylla) and barnyardgrass (Echinochloa crus-galli). The crude ethanolic extract was prepared from 100 g of dry plant material dissolved in 500 ml of ethanol. The extracts were solubilized in a buffer solution containing dimethyl sulfoxide (DMSO) at concentrations of 10.0, 7.5, 5.0 and 2.5 mg mL-1. The effect of these extracts was compared with herbicide oxyfluorfen in bioassays. The ethanolic extracts of S. lethalis leaves and stems inhibited the germination and seedling growth of barnyardgrass and wild poinsettia in a concentration-dependent manner. The reduction in the root length of E. heterophylla seedlings might be attributed to the reduced elongation of metaxylem cells. The phytotoxicity of the extracts ranged according to the receptor species, and for some variables, the inhibitory effect was similar, and even superior, to that of the commercial herbicide. Thus, S. lethalis extracts might be a promising alternative for sustainable weed management.