Development of a flotation method for preconcentration-separation of trace amounts of some metal ions in plant tissues prior to their FAAS determinations

An efficient flotation method based on the combination of flame atomic absorption spectrometry (FAAS) and separation and preconcentration step for determination of Cr3+, Cu 2+, Co2+, Ni2+, Zn2+, Cd 2+, Fe3+ and Pb2+ ions in various real samples by the possibility of applying bis(2-hydroxyacetophenone)-1,4-butanediimine (BHABDI) as a new collector was studied. The influence of pH, amount of BHABDI as collector, sample matrix, type and amount of eluting agent, type and amount of surfactant as floating agent, ionic strength and air flow rates i.e. variables affecting the efficiency of the extraction system was evaluated. It is ascertained that metal ions such as iron can be separated simultaneously from matrix in the presence of 0.012 mM ligand, 0.025% (w/v) of CTAB to a test sample of 750 mL at pH 6.5. These ions can be eluted quantitatively with 6 mL of 1.0 mol L-1 HNO3 in methanol which lead to the enrichment factor of 125. The detection limits for analyte ions were in the range of 1.3-2.4 ng mL-1. The method has been successfully applied for determination of trace amounts of ions in various real samples.

Sowing depths of brachiaria in intercropping with corn in no tillage planting

The forage production in crop-livestock integration is critical both for formation of straw for no tillage planting and food for livestock farm. The experiment was conducted in the autumn/winter of 2009 and 2010, in the city of Selvíria -state of Mato Grosso do Sul -MS, Brazil, at Experimental Station of FEIS/UNESP. The objective was to evaluate the optimal depth for deposition of seeds of two Brachiaria species intercropped with corn with emphasis on grain yield and straw. The experimental design was a randomized block design in a factorial scheme 3 x 3, with four replications. The main treatments were two species of Brachiaria (Urochloa brizantha "Marandú" and Urochloa ruziziensis), which seeds were mixed with corn fertilizer and a control treatment (without intercropping). Secondary treatments consisted of three depths (8; 10 and 16 cm) in the deposition of fertilizer (in the consortium and the control treatments). The intercropping corn with Brachiaria produced similar amounts of straw. The straw total production was higher when intercropped and decreased with depth. The consortium with U. ruziziensis provided higher grain yield of corn in relation to U. brizantha, in 2010. The sowing depth of forages did not affect corn yield.

Corn production for silage intercropped with forage in the farming-cattle breeding integration

Corn cropping for silage, due to the plant material exportation, intercropping with forage provides greater ground cover and straw formation for the Direct Planting System (DPS) continuity. The objective of this study was to evaluate corn production for silage in DPS intercropped with four forages (Urochloa brizantha cv. Marandu, U. ruziziensis cv. Ruziziensis, Panicum maximum cv. Tanzânia and P. maximum cv. Áries). We applied three sowing methods (in row together with corn fertilizer; by no-till sowing simultaneously to corn sowing and at V4 corn stage) and corn without intercropping. The experiment was conducted in autumn/ winter of 2010, in Selvíria - MS, in a randomized block design in factorial arrangement (4 x 3 + 1) and 4 replications. For corn, we evaluated plant height, basal stem diameter, initial and final stand and silage production and for forage dry matter production. Morphological characteristics and corn yield were not affected by intercropping when compared to sole corn crop. Forage dry matter production sown in corn row with fertilizer is a highlight, which in addition to providing greater productivity, harnesses the operation during sowing.

Sheep poisoning by Panicum dichotomiflorum in northeastern Brazil

Different species of Panicum, including P. dichotomiflorum,have been reported as a cause of photosensitization in sheep, horses, cattle and goats. An outbreak of hepatogenous photosensitization occurred in 3 flocks of hair sheep in the Brazilian semiarid region. Eighty one out of 365 sheep were affected and 39 died. The main affected animals were nursing lambs and sheep younger than one year old. Donkeys, goats and cattle grazing in the same pasture were not affected. Clinical signs were edema of the head, followed by dermatitis, mainly in the face, ears, and croup, ocular discharge, corneal opacity with blindness, and redness of the coronary band and hoof. At necropsy of one affected lamb the liver was yellowish. Upon histologic examination scattered necrotic hepatocytes were observed in the liver and focal areas of necrosis of myocytes appeared in the heart. Samples of P. dicotomiflorum were analyzed by TLC and those containing saponins were isolated by HPLC using RP-C18 column and eluted with a mixture of MeOH and H2O. The isolated compounds were submitted to ¹H and 13C NMR spectroscopy. Reactions were positive to furostanol saponins with the same Rf of the standard protodioscin (0.21) and methylprotodioscin (0.32). The spectroscopic results indicated a mixture of (25R)- and (25S)-protodioscin isomers in a proportion of 3:1, and methylprotodioscin.

Performance of dairy heifers raised on Xaraés palissadgrass (Brachiaria brizantha cv. Xaraés) pasture supplemented with two types of mineral mixture

The effects of selective (e.g. mineral supplement formulated on the basis of nutritional and clinical examination of the herd) or commercial mineral supplementation of crossbred dairy heifers (Holstein-Mantiqueira), on daily weight gain (DWG), body condition score (BCS), age at first mating (AFM) and the intake of mineral mixture (IMM) managed on Xaraés palissadgrass pasture (Brachiaria brizantha cv. Xaraés) were evaluated from February 2006 to March 2008. Structural characteristics, forage allowance and nutritional value of Xaraés palissadgrass were also evaluated. The structural variables of pasture, allowances and nutritive value of forage, besides the DWG were only affected (p<0.05) by season, with highest results for spring and summer. Throughout the experimental period (730 days), no sign of mineral deficiency was detected in heifers that ingested the selective supplement. The DWG and AFM were not influenced by the mineral mixture offered (0.52 and 0.33 kg/day during the spring/summer and in fall/winter for DWG, respectively, and 813 days of AFM). The IMM was higher for commercial mineral mixture than for selective (61.6 and 51.0g/day respectively). The BCS was slightly higher for selective (3.17) than for the commercial (3.02) mineral mixture. Thus, the selective mineral supplement was a strategy to prevent mineral deficiency in this herd and resulted in similar performance at lower costs (e.g. due to less IMM) of crossbred heifers when compared with the commercial mineral supplementation.

Potencial alelopático de soluções de solo cultivado com Brachiaria brizantha: efeitos sobre a germinação de gramíneas forrageiras e plantas daninhas de pastagens

O objetivo desta pesquisa foi obter informações que possam favorecer o entendimento da atuação dos compostos alelopáticos sobre o banco de sementes do solo e sobre a seleção de plantas invasoras e indicar espécies que favoreçam a renovação dos pastos. Assim, soluções de solo de uma área de pastagem de Brachiaria brizantha cv. Marandu formada há mais de cinco anos foram extraídas para avaliar os efeitos alelopáticos dessa espécie sobre a germinação, a dormência de sementes e o vigor de plântulas de B. brizantha cv. Marandu, Panicum maximum cv. Tanzânia, Sida rhombifolia e Peschiera fuchsiaefolia. O experimento foi instalado e conduzido no Laboratório de Matologia do Departamento de Produção Vegetal da FCA/UNESP - campus de Botucatu-SP. O substrato de germinação foi umedecido com 12 mL dos seguintes tratamentos: solução do solo de uma área cultivada com B. brizantha; solução do solo de uma área sem B. brizantha (mata nativa); água destilada; solução de polietilenoglicol com potencial osmótico idêntico ao da solução do solo sob B. brizantha; e solução de polietilenoglicol com potencial osmótico idêntico ao da solução de solo de uma mata nativa. A porcentagem de sementes normais, mortas, anormais e dormentes de B. brizantha não foi influenciada por nenhuma das soluções testadas, o que evidencia a não-ocorrência de efeito auto-alelopático. Foram verificados possíveis efeitos alelopáticos negativos sobre a porcentagem e velocidade de germinação de P. maximum cv. Tanzânia e sobre o crescimento radicular de S. rhombifolia. Esta última também se mostrou sensível ao efeito alelopático da solução de solo de mata, que promoveu redução no crescimento radicular. P. fuchsiaefolia não foi afetada por nenhuma das soluções testadas.

Microwave-assisted solvent extraction and analysis of shikimic acid from plant tissues

A better method for determination of shikimate in plant tissues is needed to monitor exposure of plants to the herbicide glyphosate [N-(phosphonomethyl)glycine] and to screen the plant kingdom for high levels of this valuable phytochemical precursor to the pharmaceutical oseltamivir. A simple, rapid, and efficient method using microwave-assisted extraction (MWAE) with water as the extraction solvent was developed for the determination of shikimic acid in plant tissues. High performance liquid chromatography was used for the separation of shikimic acid, and chromatographic data were acquired using photodiode array detection. This MWAE technique was successful in recovering shikimic acid from a series of fortified plant tissues at more than 90% efficiency with an interference-free chromatogram. This allowed the use of lower amounts of reagents and organic solvents, reducing the use of toxic and/or hazardous chemicals, as compared to currently used methodologies. The method was used to determine the level of endogenous shikimic acid in several species of Brachiaria and sugarcane (Saccharum officinarum) and on B. decumbens and soybean (Glycine max) after treatment with glyphosate. The method was sensitive, rapid and reliable in all cases.

Capacidade competitiva do jatobá com adubos verdes, forrageiras e plantas daninhas

O estudo do consórcio entre espécies anuais e perenes representa uma ferramenta importante no processo de implantação e manejo florestal. O objetivo deste trabalho foi avaliar a capacidade competitiva do jatobá (Hymenaea courbaril) com espécies de adubos verdes, forrageiras e plantas daninhas quanto à alocação de matéria seca, área foliar e concentração de nutrientes. Foram conduzidos dois experimentos, sendo os tratamentos compostos pela combinação de mudas de jatobá, desenvolvendo-se isoladamente ou em competição com cada uma das seguintes espécies: Brachiaria humidicola, Brachiaria brizantha, Brachiaria decumbens, Panicum maximum, Cajanus cajan, Canavalia ensiformis e Mucuna aterrima (experimento 1) e Bidens pilosa, Cenchrus echinatus, Euphorbia heterophylla, Lolium multiflorum e Solanum americanum (experimento 2), mais o cultivo de cada planta daninha e consorte isolada. Após convivência por 60 dias, as plantas foram coletadas para avaliação de matéria seca, área foliar e teor de nutrientes. Observou-se que a competição entre as plantas não promoveu alterações na produção de matéria seca ou área foliar do jatobá. Tendo em vista o exposto, verifica-se que a capacidade competitiva do jatobá não é afetada pela presença das espécies de adubos verdes e forrageiras, possibilitando convivência em fase inicial de desenvolvimento. Quanto à convivência das plantas daninhas com o jatobá, observou-se efeito positivo no acúmulo de nutrientes por estas.

Brachiaria ruziziensis and herbicide on the yield of upland rice

The correct management of cover crops in no-tillage aims to obtain greater benefits with its introduction in agricultural systems. The use of forage species such as Brachiaria, due to the large amount of biomass and for persisting longer on the ground, has become a good option. In this sense, an important point is the time interval between the cover desiccation with glyphosate and planting of rice in order to obtain higher operating income in the sowing, greater availability of nutrients, greater presence of straw on the soil surface, and lower release of possible allelopathic substances. The objective of this study was to assess the effect of the management of B. ruziziensis, with or without herbicide, preceding the crop planting, in the yield components and yield of rice. The trial was conducted in greenhouse conditions and consisted of three types of management of B. ruziziensis before sowing rice. B. ruziziensis showed linear growth and the presence of large amounts of its dry matter biomass on the soil surface was detrimental to rice yield; B. ruziziensis management close to planting caused a significant reduction in rice yield; the management of B. ruziziensis with herbicides should be conducted with more than 20 days before planting rice; the management of B. ruziziensis for the removal of its leaves with or without herbicide should be carried out with 10 or more days before planting rice, and the correct management of B. ruziziensis at the right time allowed for significant increases in the rice yield.

Levels of nutrients and grain yield of maize intercropped with signalgrass (Brachiaria) in different arrangements of plants

Competition between maize and signalgrass can economically cripple the intercropping by the reduced yield of maize and dry matter content of the forage. In seeking to define plant arrangements which make this system more efficient, this research was held with the objective of assessing the effects of interference of densities of signalgrass (Urochloa Brizantha) on nutrition and on maize grain yield. Two field experiments were conducted in a randomized block design with four replications. Treatments were arranged similarly in both experiments, in a 2 x 4 factorial design, the first factor being the dose of Nicosulfuron herbicide applied (0 and 8 g ha-1) and the second factor being the forage seeding rates (0, 2, 4 and 6 kg of seeds per hectare). The interference of signalgrass reduced foliar nitrogen, potassium and phosphorus content in maize plants intercropped with the forage. Higher values of grain yield were observed with the reduction of the spacing and the application of the recommended herbicide underdose (8 g ha-1). It was concluded that, regardless of the seeding density of U. Brizantha, reducing the maize seeding inter-rows spacing, combined with the application of an underdose of Nicosulfuron, caused a positive effect by reducing the initial forage growth, resulting in less interference of Urochloa brizantha on nutrient uptake by the maize plants and grain yield of the crop.

The Allelopathic Effect of Eucalyptus Leaf Extract on Grass Forage Seed

This study aimed to evaluate the allelopathic effect of various concentrations of an aqueous extract of eucalyptus leaves on Urochloa decumbens and Panicum maximum seeds. The extract was prepared from Eucalyptus urograndis leaves that were milled and mixed with distilled water in a 1:9 milled leaves: water ratio to obtain an extract with a defined concentration of 100%. In addition, dilutions of 50%, 25% and 12.5% were prepared, and a 0% dilution was used as a control. The experiment followed a completely randomized design, with four replicates, each of 50 seeds of U. decumbens and 50 seeds of P. maximum, arranged on filter paper moistened with each concentration of extract in a Gerbox plastic box. The results demonstrated the allelopathic potential of E. urograndis aqueous extracts applied to the seeds of U. decumbens and P. maximum. The 50% and 100% concentrations of leaf extract most strongly inhibited the germination, vigor and seedling growth of U. decumbens and P. maximum. The germination speed index and the root length were the characteristics that were most affected by the potentially allelopathic substances contained in the eucalyptus extracts at all concentrations.

Diurnal variations of non-structural carbohydrates in vegetative tissues of Melinis minutiflora, Echinolaena inflexa and Lolium multiflorum (Poaceae)

Echinolaena inflexa (Poir.) Chase is a native C3 grass species with high biomass production and abundance in the cerrado comparable to Melinis minutiflora Beauv., a C4 forage grass of African origin, widespread in the cerrado, displacing some native herbaceous species. These species accumulate mainly starch, differently of Lolium multiflorum Lam. ssp. italicum Beck cv. Lema (ryegrass), a C3 temperate forage grass that accumulates fructans as the major storage carbohydrate in the vegetative tissues. In the present study, we analyzed diurnal variations in the content and composition of non-structural carbohydrates (NSC) in the aerial vegetative parts of the three species. Results showed similarity in water-soluble carbohydrates (WSC) and starch contents between Melinis minutiflora and Echinolaena inflexa, with the exotic grass accumulating higher amounts of NSC. However, the levels of sucrose, glucose and fructose were 10 fold higher in Lolium multiflorum, that accumulates fructan as the main storage carbohydrate. Although the contents of WSC and starch were similar in the tropical species E. inflexa and M. minutiflora, their diurnal variations were different and could be related to starch metabolism, especially in M. minutiflora.

Blood flow measurements in rats using four color microspheres during blockade of different vasopressor systems

The use of colored microspheres to adequately evaluate blood flow changes under different circumstances in the same rat has been validated with a maximum of three different colors due to methodological limitations. The aim of the present study was to validate the use of four different colors measuring four repeated blood flow changes in the same rat to assess the role of vasopressor systems in controlling arterial pressure (AP). Red (150,000), white (200,000), yellow (150,000), and blue (200,000) colored microspheres were infused into the left ventricle of 6 male Wistar rats 1) at rest and 2) after vasopressin (aAVP, 10 µg/kg, iv), 3) renin-angiotensin (losartan, 10 mg/kg, iv), and 4) sympathetic system blockade (hexamethonium, 20 mg/kg, iv) to determine blood flow changes. AP was recorded and processed with a data acquisition system (1-kHz sampling frequency). Blood flow changes were quantified by spectrophotometry absorption peaks for colored microsphere components in the tissues evaluated. Administration of aAVP and losartan slightly reduced the AP (-5.7 ± 0.5 and -7.8 ± 1.2 mmHg, respectively), while hexamethonium induced a 52 ± 3 mmHg fall in AP. The aAVP injection increased blood flow in lungs (78%), liver (117%) and skeletal muscle (>150%), while losartan administration enhanced blood flow in heart (126%), lungs (100%), kidneys (80%), and gastrocnemius (75%) and soleus (94%) muscles. Hexamethonium administration reduced only kidney blood flow (50%). In conclusion, four types of colored microspheres can be used to perform four repeated blood flow measurements in the same rat detecting small alterations such as changes in tissues with low blood flow.

Symmetric pollen mitosis I and suppression of pollen mitosis II prevent pollen development in Brachiaria jubata (Gramineae)

Microsporogenesis and pollen development were analyzed in a tetraploid (2n = 4x = 36) accession of the forage grass Brachiaria jubata (BRA 007820) from the Embrapa Beef Cattle Brachiaria collection that showed partial male sterility. Microsporocytes and pollen grains were prepared by squashing and staining with 0.5% propionic carmine. The meiotic process was typical of polyploids, with precocious chromosome migration to the poles and laggards in both meiosis I and II, resulting in tetrads with micronuclei in some microspores. After callose dissolution, microspores were released into the anther locule and appeared to be normal. Although each microspore initiated its differentiation into a pollen grain, in 11.1% of them nucleus polarization was not observed, i.e., pollen mitosis I was symmetric and the typical hemispherical cell plate was not detected. After a central cytokinesis, two equal-sized cells showing equal chromatin condensation and the same nuclear shape and size were formed. Generative cells and vegetative cells could not be distinguished. These cells did not undergo the second pollen mitosis and after completion of pollen wall synthesis each gave rise to a sterile and uninucleate pollen grain. The frequency of abnormal pollen mitosis varied among flowers and also among inflorescences. All plants were equally affected. The absence of fertile sperm cells in a considerable amount of pollen grains in this accession of B. jubata may compromise its use in breeding and could explain, at least in part, why seed production is low when compared with the amount of flowers per raceme.

Imagens digitalizadas na interpretação do teste de tetrazólio em sementes de Brachiaria brizantha

As espécies forrageiras, principalmente dos gêneros Brachiaria e Panicum, desempenham importante papel nas regiões pecuárias e têm auxiliado o desenvolvimento da indústria de sementes no Brasil, o qual se transformou em maior produtor, consumidor e exportador de sementes de gramíneas forrageiras tropicais. O objetivo desse trabalho foi avaliar lotes de sementes de Brachiaria brizantha 'Marandu' por meio do teste de tetrazólio conduzido mediante dois procedimentos. Foram utilizados 5 lotes de sementes de Brachiaria brizantha 'Marandu' de origens diferentes, avaliados pelo teste de germinação, após escarificação ou não com ácido sulfúrico, e pelo teste de tetrazólio com duas formas de avaliação: método convencional, sob estéreo microscópio, e análise de imagens digitalizadas, obtidas de sementes agrupadas em placa de vidro de alta transparência, com definição de 1200 dpi. O experimento foi conduzido em delineamento inteiramente casualizado. A avaliação de lotes de sementes de Brachiaria brizantha 'Marandu', conduzida por meio do teste de tetrazólio, com observação das imagens digitalizadas das sementes é equivalente à efetuada sob estéreo microscópio.