Efeitos alelopáticos de extratos aquosos de Crinum americanum L.

Investigamos a ação de extratos aquosos de Crinum americanum L. sobre a germinação e crescimento inicial de Lactuca sativa L., Sesamum indicum L. e Raphanus sativus L. e das espécies invasoras: Echinochloa crusgalli (L.) P. Beauv., Ipomoea grandifolia (Dammer) O'Donell e Bidens pilosa L. Preparamos os extratos aquosos a partir de exemplares de um estuário cego em Caraguatatuba, São Paulo. Montamos controles osmóticos com PEG 6000. Analisamos os dados de germinação com Kruskal-Wallis e pós-teste de Dunn, e os dados de crescimento inicial com ANOVA e pós-teste de Tukey. Para todas as espécies testadas obtivemos diminuição na porcentagem e no crescimento inicial e aumento do tempo médio da germinação. O controle osmótico indicou pouca influência da osmolaridade na germinação. Concluímos que C. americanum possui potencial alelopático sobre as espécies testadas.

Studies on ATP-diphosphohydrolase nucleotide-binding sites by intrinsic fluorescence

Potato apyrase, a soluble ATP-diphosphohydrolase, was purified to homogeneity from several clonal varieties of Solanum tuberosum. Depending on the source of the enzyme, differences in kinetic and physicochemical properties have been described, which cannot be explained by the amino acid residues present in the active site. In order to understand the different kinetic behavior of the Pimpernel (ATPase/ADPase = 10) and Desirée (ATPase/ADPase = 1) isoenzymes, the nucleotide-binding site of these apyrases was explored using the intrinsic fluorescence of tryptophan. The intrinsic fluorescence of the two apyrases was slightly different. The maximum emission wavelengths of the Desirée and Pimpernel enzymes were 336 and 340 nm, respectively, suggesting small differences in the microenvironment of Trp residues. The Pimpernel enzyme emitted more fluorescence than the Desirée apyrase at the same concentration although both enzymes have the same number of Trp residues. The binding of the nonhydrolyzable substrate analogs decreased the fluorescence emission of both apyrases, indicating the presence of conformational changes in the neighborhood of Trp residues. Experiments with quenchers of different polarities, such as acrylamide, Cs+ and I- indicated the existence of differences in the nucleotide-binding site, as further shown by quenching experiments in the presence of nonhydrolyzable substrate analogs. Differences in the nucleotide-binding site may explain, at least in part, the kinetic differences of the Pimpernel and Desirée isoapyrases.

Mycoflora and aflatoxigenic species in derivatives of milled rice

Thirty samples of rough rice stored for 6, 12 and 24 months in government authorized warehouses of the state of Rio Grande do Sul, Brazil, were simultaneously collected. After milling of the product, 90 samples (30 of polished rice, 30 of rice bran and 30 of rice hull) were evaluated for their mycoflora, aflatoxigenic species and aflatoxin contamination. The following fungi, listed in decreasing order of frequency, were isolated on Potato-Dextrose Agar: Aspergillus spp., Nigrospora spp., Penicillium spp.; Fusarium spp.; Mucor spp.; Cladosporium spp.; Trichosporon spp. and non-sporulated fungi. The degree of fungal contamination (colony forming units per gram of product) was lowest in polished rice, increasing progressively in samples of rice bran and rice hull. Among the Aspergillus species, A. flavus and A. candidus were isolated most frequently. Of the A. flavus isolates, 52.6% strains were found to be toxigenic and produced only Group B aflatoxins. Analysis of the 90 samples did not reveal the presence of aflatoxins in the rice derivatives.

Efeito do cloreto de sódio na produção de proteínas (Saccharomyces cerevisiae) em fermentação semi-sólida

Estudou-se o efeito do cloreto de sódio sobre a produção de biomassa e proteínas extracelulares totais, durante o cultivo de Saccharomyces cerevisiae. A levedura foi desenvonvida em fermentador de leito fluidizado, com vazão de ar de 70L/min, temperatura de 33° C, e umidade relativa de 99-100%. Foi utilizado substrato semi-sólido de batatas, previamente hidrolizado, acrescido de cloreto de sódio 0,6M. O crescimento celular foi monitorado por densidade óptica à 595 nm. Observou-se, como resultado, que a adição de cloreto de sódio 0,6M induziu um aumento de 36,86% na produção de proteínas extracelulares totais, mas inibiu o crescimento celular em 27,62% quando os meios com e sem cloreto de sódio foram testados. A produção máxima de biomassa, tanto para os experimentos com adição de cloreto de sódio quanto para o sem adição, ocorreu no período de 7 a 9 horas de fermentacão, enquanto que a produção de proteínas extracelulares totais, independentemente da adição do sal, ocorreu durante o período de 9 a 12 horas de fermentação. As velocidades específicas máximas de crescimento foram de 0,350/h para os experimentos com sal, e de 0,339/h para aqueles sem a adição do sal. A combinação de alta vazão de ar e a presença de cloreto de sódio 0,6M na fermentação parece não ter tido efeito sobre a duração da fase lag na curva de crescimento celular de Saccharomyces cerevisiae.

Comportamento de óleos vegetais em frituras descontínuas de produtos pré-fritos congelados

Este estudo teve como objetivo avaliar algumas alterações de óleos vegetais (girassol, soja e milho), utilizado em sucessivas frituras de produtos pré-fritos congelados (batata palito e produto cárneo empanado (snacks). As frituras dos produtos foram conduzidas em fritadeira doméstica e com as seguintes condições controladas: temperatura de 180°C, relação superfície/volume de 0,3 cm-1 e tempo total de aquecimento de 12 h. Nas amostras dos óleos procederam-se as determinações analíticas: compostos polares totais, dienos conjugados, índice de ácido tiobarbitúrico (TBA) e medida da estabilidade oxidativa. Os resultados, em duplicata, obtidos das determinações analíticas foram submetidos às análises de variância, empregando um esquema fatorial, no delineamento inteiramente casual, de modo a determinar a influência dos fatores (produtos, óleos e tempos de fritura) sobre as alterações nos óleos. Os óleos vegetais utilizados nas frituras dos snacks apresentaram menores alterações do que os óleos utilizados para fritura das batatas. Os resultados mostraram que os óleos estudados, apesar das diferenças na composição em ácidos graxos, não apresentaram, em nenhuma análise, valores acima dos limites recomendados em alguns países para o descarte de óleos, independentemente do tipo de produto frito e tempo de aquecimento.

Composição química de tubérculos de batata para processamento, cultivados sob diferentes doses e fontes de potássio

A batata é considerada um dos poucos alimentos capazes de nutrir a crescente população mundial por ser fonte de energia e conter elevado teor de proteínas, vitaminas e minerais. Porém os teores desses compostos sofrem influência de diversos fatores como: cultivar utilizada, condições edafoclimáticas, safra, colheita e armazenamento. Assim, o presente trabalho teve como objetivo determinar a composição química de batatas (cvs. Atlantic, Asterix, Innovator e Shepody), cultivadas em quatro doses (0, 120, 360 e 1080 kg K2O.ha-1) e duas fontes de potássio (KCl e K2SO4). As amostras foram provenientes do município de Fazenda Rio Grande/PR, cultivadas na safra das águas. Foram realizadas as seguintes determinações: vitamina C, umidade, proteínas, lipídios, cinzas, carboidratos, energia, amido e potássio. A cv. Atlantic apresentou os maiores teores médios de cinzas (0,93%) e potássio (528,80 mg.100 g-1); a cv. Asterix, a maior umidade (81,47%); a cv. Innovator, os maiores teores de proteínas (2,25%), lipídios (0,06%), carboidratos (17,72%), energia (80,40 kcal.100 g-1) e amido (16,45%); e a cv. Shepody obteve a maior quantidade de vitamina C (31,01 mg.100 g-1). Pode-se concluir que a composição química das batatas é dependente da cultivar e da adubação potássica (dose e fonte) empregada.

Effect of enzymatic hydrolysis on some physicochemical properties of root and tuber granular starches

Enzymatic hydrolysis of granular starch is an important tool to provide information about granule structure. Cassava, sweet potato, Peruvian carrot, and potato starches were hydrolyzed by bacterial α-amylase at 37 °C for 48 hours, and the physicochemical properties of the residues from hydrolysis were determined. Cassava starch was the most susceptible to enzyme displaying 20.9% of hydrolysis, whereas potato starch was the most resistant with 5.9%. The granule average size varied from 10.8 to 23.4 μm for Peruvian carrot and potato starches, respectively. With the use of SEM, a smooth granule surface was observed for all native starches. Cassava and sweet potato starches displayed an A-type X-ray diffraction pattern, while Peruvian carrot and potato starches showed a B-type pattern. After hydrolysis, cassava, sweet potato, and Peruvian carrot starches showed some well degraded granules, whereas potato starch presented a slight sign of degradation. The amylose content of the starches decreased with hydrolysis for cassava, sweet potato, and Peruvian carrot starches and was kept unchanged for the potato starch. As expected, intrinsic viscosity and pasting properties decreased for all hydrolyzed starches. There is no difference between thermal properties of native and hydrolyzed starches. These results suggested that hydrolysis occurred in amorphous and crystalline areas of the granules. The B type diffraction pattern in conjunction with the big granule size of the potato starch may have contributed to the greatest resistance of this starch to hydrolysis.

Sugarcane starch: quantitative determination and characterization

Starch is found in sugarcane as a storage polysaccharide. Starch concentrations vary widely depending on the country, variety, developmental stage, and growth conditions. The purpose of this study was to determine the starch content in different varieties of sugarcane, between May and November 2007, and some characteristics of sugarcane starch such as structure and granules size; gelatinization temperature; starch solution filterability; and susceptibility to glucoamylase, pullulanase, and commercial bacterial and fungal α-amylase enzymes. Susceptibility to debranching amylolytic isoamylase enzyme from Flavobacterium sp. was also tested. Sugarcane starch had spherical shape with a diameter of 1-3 µm. Sugarcane starch formed complexes with iodine, which showed greater absorption in the range of 540 to 620 nm. Sugarcane starch showed higher susceptibility to glucoamylase compared to that of waxy maize, cassava, and potato starch. Sugarcane starch also showed susceptibility to debranching amylolytic pullulanases similar to that of waxy rice starch. It also showed susceptibility to α-amylase from Bacillus subtilis, Bacillus licheniformis, and Aspergillus oryzae similar to that of the other tested starches producing glucose, maltose, maltotriose, maltotetraose, maltopentose and limit α- dextrin.

Optimization of extrusion process for production of nutritious pellets

A blend of 50% Potato Starch (PS), 35% Quality Protein Maize (QPM), and 15% Soybean Meal (SM) were used in the preparation of expanded pellets utilizing a laboratory extruder with a 1.5 × 20.0 × 100.0 mm die-nozzle. The independent variables analyzed were Barrel Temperature (BT) (75-140 °C) and Feed Moisture (FM) (16-30%). The effect of extrusion variables was investigated in terms of Expansion Index (EI), apparent density (ApD), Penetration Force (PF) and Specific Mechanical Energy (SME), viscosity profiles, DSC, crystallinity by X-ray diffraction, and Scanning Electronic Microscopy (SEM). The PF decreased from 30 to 4 kgf with the increase of both independent variables (BT and FM). SME was affected only by FM, and decreased with the increase in this variable. The optimal region showed that the maximum EI was found for BT in the range of 123-140 °C and 27-31% for FM, respectively. The extruded pellets obtained from the optimal processing region were probably not completely degraded, as shown in the structural characterization. Acceptable expanded pellets could be produced using a blend of PS, QPM, and SM by extrusion cooking.

6-n-propyltiouracil (PROP) taster status in Brazilian adults

The objective of this study was to determine PROP (6-n-propyltiouracil) taster status in adults and its relationship with anthropometric variables and pleasantness of sugar, salt, and fat. A total of 123 subjects rated the intensity of PROP and sodium cloride (NaCl) solutions using the labeled magnitude scale. For pleasantness evaluation, it was used concentrated orange juice (sugar) and mashed potato (salt and fat). The subjects were classified as non-tasters (n = 35), medium-tasters (n = 33) and super-tasters (n = 55). In this study, no relationship was found between PROP taster status and age, sex, weight, body mass index, and pleasantness. Although genetic markers may influence the degree of liking of certain foods, one must consider that the mechanisms influencing eating behavior in humans are complex, and that psychological, social, and economic factors play a key role in response to food.

Thermal inactivation of polyphenoloxidase and peroxidase in Jubileu clingstone peach and yeast isolated from its spoiled puree

The thermal inactivation of yeast isolated from spoiled Jubileu peach puree and that of polyphenoloxidase (PPO) and peroxidase (POD) in cv. Jubileu, which is widely cultivated in southern Rio Grande do Sul state, Brazil, were studied. PPO and POD were extracted using the protein powder method and submitted to partial purification by precipitation followed by dialysis. The enzymatic activity was determined measuring the increase in absorbance at 420 nm for PPO and 470 nm for POD. The yeast used in this investigation was isolated from spoiled Jubileu peach puree at 22 °Brix, with total initial microbial count of 22 × 10² UFCmL- 1. Stock cultures were maintained on potato dextrose agar (PDA) slants at 4 °C and pH 5 for later use for microbial growth. In all cases, kinetic analysis of the results suggests that the thermal inactivation was well described by a first-order kinetic model, and the temperature dependence was significantly represented by the Arrhenius law. Both enzymes were affected by heat denaturation, and PPO was more thermostable. PPO was also more thermosTable than the yeast isolated from peach puree. The D60-values were 1.53 and 1.87 min for PPO and yeast isolated from spoiled Jubileu peach puree, respectively.

Detection, transmission and pathogenicity of fungi on Blepharocalyx salicifolius (H.B.K.) Berg. seeds

The objective of this study was to identify the fungi associated with the fruit and seeds of Blepharocalyx salicifolius and verify their transmission and pathogenicity to seeds and seedlings. Fungal identification on seeds was made using the blotter test and potato-dextrose-agar but only the blotter test was used for fruit. Fungal transmission to seedlings was evaluated using four replications of 50 seeds planted in vermiculite. The pathogenicity of the fungi, Colletotrichum sp., Curvularia sp., Cladosporium sp. Pestalotia sp. and Macrophomina sp. was tested. Potentially pathogenic and saprophytic fungi were found on the fruits and seeds. The transmission of Cladosporium sp. from seeds to seedlings was verified, and Cladosporium sp. Pestalotia sp. and Macrophomina sp. were found to be pathogenic to B. salicifolius seedlings.

Survey, survival and control of Alternaria alternata in wheat seeds

The fungus Alternaria alternata was quantified in 75 wheat seed samples collected from three different regions of southern Brazil for Cropping and Use Value (CUV) I, II and III. Fungal presence was evaluated in two hundred disinfested seeds per sample before sowing in a potato-dextrose-agar medium + antibiotic (PDA+A). Fungus survival was evaluated every 45 days for 180 days for three seed batches from six wheat cultivars stored in propylene bags in a storehouse, with air temperature varying between 18 to 22 °C and relative air humidity around 60%. The efficacy of carboxin+thiram, difenoconazol, thiram, triadimenol, triticonazol and triticonazol + iprodione fungicides to control A. alternata was determined. A. alternata was detected in all the samples with an incidences of 39.6 %, 38.8% and 35.9% for the CUV I, CUV II and CUV III regions, respectively. The highest mean incidence of the fungus was found in the CUV I region, the coolest and most humid, and was significantly different from the other two regions. The average reduction in A. alternata viability in the wheat cultivar seeds was 49.5% during the 180 days of storage (inter-harvest period), demonstrating that infected seeds are the primary inoculum source for the fungus. The triticonazol + iprodione fungicide mixture efficiently controls A. alternata.

Incidence of resistance to benzimidazole fungicides in a field population of Venturia inaequalis /

The allele-specific polymerase chain reaction (PCR) was used to screen for the presence of benomyl resistance, and to characterize their levels and frequencies in field populations of Venturia inaequalis during two seasons. Three hundred isolates of V. inaequalis were collected each season from infected leaves of MalusX domestica. Borkh c.v. Mcintosh. The trees used were sprayed in the year prior to collection with five applications of benomyl, its homologue Azindoyle, or water. Monoconidial isolates of V. inaequalis were grown on 2% potato dextrose agar (PDA) for four weeks. Each isolate was taken from a single lesion from a single leaf. Total genomic DNA was extracted from the four week old colonies of V. inaequalis, prepared and used as a template in PCR reactions. PCR reactions were achieved by utilizing allele-specific primers. Each primer was designed to amplify fragments from a specific allele. Primer Vin was specific for mutations conferring the ben^^"^ phenotype. It was expected to amplify a 171 bp. DNA fragment from the ben^"^ alleles only. Primers BenHR and BenMR were specific for mutations conferring the ben"" and ben'^'' phenotypes, respectively. They were expected to amplify 172 bp. and 165 bp. DNA fragments from the ben"" and ben"^" alleles, respectively. Of the 953 isolates tested, 414 (69.9%) were benomyl sensitive (ben^) and 179 (30.1%) were benomyl resistant. All the benomyl resistant alleles were ben^"", since neither the ben"" nor the ben"" alleles were detected. Frequencies of benomyl resistance were 23%, 24%, and 23% for the 1997 collections, and were 46%, 26% and 38% for the 1998 collections for benomyl, Azindoyle and water treatments, respectively. Growth assay was performed to evaluate the applicability of using PCR in monitoring benomyl resistance in fungal field populations. Tests were performed on 14 isolates representing the two phenotypes (ben^ and ben^"'' alleles) characterized by PCR. Results of those tests were in agreement with PCR results. Enzyme digestion was also used to evaluate the accuracy and reliability of PCR products. The mutation associated with the ben^"'' phenotype creates a unique site for the endonuclease enzyme Bsh^236^ allowing the use of enzyme digestion. Isolates characterized by PCR as ben^'^'^ alleles had this restriction site for the SsA7l2361 enzyme. The most time consuming aspect of this study was growing fungal isolates on culture media for DNA extraction. In addition, the risk of contamination or losing the fungus during growth processes was relatively high. A technique for extracting DNA directly from lesions on leaves has been used (Luck and Gillings 1 995). In order to apply this technique in experiments designed to monitor fungicide resistance, a lesion has to be homogeneous for fungicide sensitivity. For this purpose, PCR protocol was used to determine lesion homogeneity. One hundred monoconidial isolates of V. inaequalis from 10 lesions (10-conidia/ lesion) were tested for their phenotypes with respect to benomyl sensitivity. Conidia of six lesions were homogeneous, while conidia of the remaining lesions were mixtures of ben^ and ben^ phenotypes. Neither the ben" nor the ben' phenotype was detected.

Sensitivities of Ontario isolates of Venturia inaequalis to metiram /

The effects of metiram (Polyram 80 DF) on the growth of Venturia inaequalis, cause of apple scab, and the degradation of metiram were examined in culture media. Samples of V. inaequalis conidia were collected from nine orchards in 1998 and six orchards in 1999 and tested for sensitivity. Samples were plated on water agar amended with metiram or mancozeb. Mean EC50 values (effective concentration of fungicide required to inhibit germination of half the conidia) for each population were calculated. The mean EC50 values for metiram ranged from 0.26 - 1.20 ^ig metiram a.i./ml, with differences (Student Newman Keul's Test (SNK), a=0.05) between populations. EC50 values for mancozeb ranged from 0.06 - 0.58 which were also different (SNK, a=0.05). Five of these populations were examined for mycelial growth sensitivity to metiram by testing 30 monoconidial isolates from each population on metiram amended potato dextrose agar. Mean EC50 values for populations were calculated and ranged from 3.44-5.94 |ig metiram/ml, and showed differences (Friedman Test, a=0.05). As the EC50 values obtained are far less than the concentrations applied in the field, results indicate that Ontario populations of V. inaequalis are still sensitive to metiram and mancozeb. The stability of metiram in PDA at 22°C was studied over a 10-day period. The initial concentration of metiram decreased by approximately 50% within the first day, and continued to decline slowly, to approximately 20% of the initial concentration. The factors possibly affecting initial metiram degradation, including agar, heat, and the use of glass or polystyrene Petri dish composition were examined. The effects from the polystyrene in the Petri dish composition were negligible, however more studies must be done to examine metiram degradation during the first 24 hours of preparation.