Proteinase inhibitors in Brazilian leguminosae

Serine proteinase inhitors, in the seeds of several Leguminosae from the Pantanal region (West Brazil), were studied using bovine trypsin, a digestive enzyme, Factor XIIa and human plasma Kallikrein, two blood clotting factors. The inhibitors were purified from Enterolobium contortisiliquum (Mr=23,000), Torresea cearensis (Mr = 13,000), Bauhinia pentandra (Mr = 20,000) and Bauhinia bauhinioides (Mr = 20,000). E. contortisiliquum inhibitor inactivates all three enzymes, whereas the T. cearensis inhibitor inactivates trypsin and Factor XSSa, but does nor affect plasma kallikrein; both Bauhinia inhibitors, on the other hand, inactivate trypsin and plasma kallikrein but only the Bpentandra inhibitor affects Factor XIIa. Ki values were calculated between 10 [raised to the power of] -7 and 10 [raised to the power of] -8 M.

Kazal-type serine proteinase inhibitors in the midgut of Phlebotomus papatasi

Sandflies (Diptera: Psychodidae) are important disease vectors of parasites of the genus Leishmania, as well as bacteria and viruses. Following studies of the midgut transcriptome of Phlebotomus papatasi, the principal vector of Leishmania major, two non-classical Kazal-type serine proteinase inhibitors were identified (PpKzl1 and PpKzl2). Analyses of expression profiles indicated that PpKzl1 and PpKzl2 transcripts are both regulated by blood-feeding in the midgut of P. papatasi and are also expressed in males, larva and pupa. We expressed a recombinant PpKzl2 in a mammalian expression system (CHO-S free style cells) that was applied to in vitro studies to assess serine proteinase inhibition. Recombinant PpKzl2 inhibited α-chymotrypsin to 9.4% residual activity and also inhibited α-thrombin and trypsin to 33.5% and 63.9% residual activity, suggesting that native PpKzl2 is an active serine proteinase inhibitor and likely involved in regulating digestive enzymes in the midgut. Early stages of Leishmania are susceptible to killing by digestive proteinases in the sandfly midgut. Thus, characterising serine proteinase inhibitors may provide new targets and strategies to prevent transmission of Leishmania.

Protective immunity induced in mice by F8.1 and F8.2 antigens purified from Schistosoma mansoni eggs

Schistosoma mansoni soluble egg antigens (SEA) were fractionated by isoelectric focusing, resulting in 20 components, characterized by pH, absorbance and protein concentration. The higher absorbance fractions were submitted to electrophoresis, and fraction 8 (F8) presented a specific pattern of bands on its isoelectric point. Protein 3 was observed only on F8, and so, it was utilized to rabbit immunization, in order to evaluate its capacity of inducing protective immunity. IgG antibodies from rabbit anti-F8 serum were coupled to Sepharose, and used to obtain the specific antigen by affinity chromatography. This antigen, submitted to electrophoresis, presented two proteic bands (F8.1 and F8.2), which were transferred to nitrocellulose membrane (PVDF) and sequenciated. The homology of F8.2 to known proteins was determined using the Basic Local Alignment Search Tool program (BLASTp). Significant homologies were obtained for the rabbit cytosolic Ca2+ uptake inhibitor, and for the bird a1-proteinase inhibitor. Immunization of mice with F8.1 and F8.2, in the presence of Corynebacterium parvum and Al(OH)3 as adjuvant, induced a significant protection degree against challenge infection, as observed by the decrease on worm burden recovered from portal system.

Retrospective study on clinical management of indolent ulcers in Boxer dogs

Indolent ulcers are superficial corneal ulcers secondary to several changes on the corneal surface. They are frequently observed in middle-aged Boxer dogs, cause pain of acute onset and requires appropriate treatment. Aiming to evaluate the efficacy of clinical managements on the rate of healing of indolent ulcers, a retrospective study was conducted (1997-2008). Results demonstrated that proteinase inhibitors were the most often prescribed medication, and its administration did not interfere on the healing rate, as well as observed in dogs that received 1% atropine, antibiotics and anti-inflammatory drugs. Healing was delayed in dogs administered orally with vitamin C, but the healing process was faster on those dogs that went through corneal debridement/cauterization. In conclusion, to know the various types of treatments seems to be fundamental for the rapid resolution of the disease. It is suggested that debridement/cauterization, administration of proteinase inhibitor eye drops, prophylactic topical antibiotics and oral vitamin C, should be considered as an effective clinical management for indolent ulcers in Boxer dogs.

Brazilian soybean Glycine max (L.) Merr. cultivars adapted to low latitude regions: seed composition and content of bioactive proteins

Among the goals of the Brazilian soybean improvement programmes, the breeding strategies for cultivars adapted to low latitudes have been included to extend crop areas and to increase production. Seeds of nine Brazilian soybean cultivars adapted to low latitudes were investigated regarding to their composition, and amino acid and antinutritional/toxic protein contents. Protein (394.5 ± 13.1 to 445.3 ± 8.0 g kg-1 dry matter) and oil (200.6 ± 1.2 to 232.3 ± 4.7 g kg-1 dry matter) contents showed low correlation to each other (r = -0.06). The total carbohydrate (141.7 ± 6.1 to 211.1 ± 15.0 g kg-1 dry matter) and ash contents (48.2 ± 4.2 to 52.2 ± 0.5 g kg-1 dry matter) were similar to data available for other soybean cultivars. All soybean cultivars presented low levels of tryptophan and sulphur amino acids. The lectin (1,152 to 147,456 HU kg-1 flour), trypsin inhibitor (34.45 ± 2.28 to 77.62 ± 2.63 g trypsin inhibited kg-1 flour), toxin (6,210 ± 134 to 34,650 ± 110 LD50 kg-1 flour) and urease (0.74 ± 0.02 to 1.22 ± 0.10 g kg¹ flour) presented variations in their contents amongst the cultivars. Compared to other soybean cultivars, urease was higher, the acute toxicity lower and the lectin and trypsin inhibitor contents similar to data available. In general, soybean cultivars showed similar biochemical composition to those developed in different geographic regions. The relevance of these findings to the agronomic features and to choice of soybean cultivars to be used as food or feed is discussed.

Effect of harvest at different times of day on the physical and chemical characteristics of vegetable-type soybean

The objective of this study was to evaluate the effect of harvest at different times of day on the chemical and physical characteristics of vegetable-type soybean BRS 267 cultivar, harvested at the R6 stage (seed development) and to compare it with that on the grains harvested at the R8 stage (maturation). The pods of the BRS 267 cultivar were harvested at the R6 stage (at 8:00 AM, 12:00 AM, and 6:00 PM), the color parameters were evaluated, and the grains were analyzed for chemical composition, activity inhibitor trypsin, phytic acid content, starch, sugars, fatty acids, and isoflavones. No differences were observed among the different harvest times in terms of the chemical constituents of vegetable-type soybean BRS 267 cultivar harvested at the R6 stage. Isoflavones content did not change with different harvest times, and the aglycone forms (daidzein, glycitein, and genistein) were found in smaller quantities at the R6 stage compared to the R8 stage. The color of the pods of soybean BRS 267 cultivar, harvested at the R6 stage did not change with different harvest times. The grains harvested at the R6 stage had lower protein content, phytic acid, and sucrose and higher levels of lipids, carbohydrates, starch, glucose, fructose, stachyose, and linolenic acids than those collected at the R8 stage. The different times of harvest did not affect the quality of the vegetable-type soybean BRS 267 cultivar harvested at stage R6. Nevertheless, it is recommended to harvest in the morning, when the temperature is milder, like other vegetables, to facilitate and optimize its marketing and in natura consumption.

Chemical composition of tempeh from soybean cultivars specially developed for human consumption

Tempeh is a food obtained by fermentation of soybean grains by the fungus Rizophus oligosporus. It is a traditional Indonesian food that presents benefits for human health protecting against diarrhea and chronic diseases. Tempeh processing includes dehulling, cooking, inoculation, and fermentation. In this study, chemical characteristics of tempeh prepared with soybean cultivars specially developed for human consumption (BRS 216, BRS 232, BRS 257, and BRS 267) were investigated. Soybean grains and tempeh obtained from these cultivars were analyzed for oil, protein, antinutrional factors, and isoflavone content. Cultivar BRS 216 presented the highest protein content in the grains (36.81%) and in tempeh (51.99%). On average, the protein content in tempeh increased 16% in relation to that of soybean grains. Isoflavone content was higher in the grains than in tempeh with significant differences among the cultivars. However, the aglycones content increased about 50% in tempeh (49.00 mg.100 g-1 on average) compared to that of raw material (soybean grains - 21.49 mg.100 g-1, on average). The content of Kunitz trypsin inhibitor (KSTI) reduced 83% in tempeh, on average, as compared to the value found in the grains. Phytic acid content was similar in both tempeh and the grains.

Heat-treatment reduces anti-nutritional phytochemicals and maintains protein quality in genetically improved hulled soybean flour

The soybean is a protein source of high biological value. However, the presence of anti-nutritional factors affects its protein quality and limits the bioavailability of other nutrients. The effect of heat-treatment, 150 ºC for 30 minutes, on hulled and hull-less soybean flour from the cultivar UFVTN 105AP on urease, trypsin inhibitor activity, protein solubility, amino acid profile, and in vivo protein quality was investigated. The treatment reduced the trypsin inhibitor activity and urease, but it did not affect protein solubility. Protein Efficiency Coefficient (PER) values of the flours were similar, and the PER of the hull-less soybean flour did not differ from casein. The Net Protein Ratio (NPR) did not differ between the experimental groups. The True Digestibility (TD) of the flours did not differ, but both were lower in casein and the Protein Digestibility Corrected Amino Acid Score (PDCCAS) was lower than the TD, due to limited valine determined by the chemical score. Therefore, the flours showed reduced anti-nutritional phytochemicals and similar protein quality, and therefore the whole flours can be used as a source of high quality protein.

Soybean growth and yield after single tillage and species mixture of cover plants

The use of cover crops is important for the agricultural crop and soil management in order to improve the system and, consequently, to increase yield. Therefore, the present study analyzed the effect of crop residues of black oat (Avena strigosa Schreb.) (BO) and a cocktail (CO) of BO, forage turnip (Raphanus sativus L.) (FT) and common vetch (Vicia sativa L.) (V) on the emergence speed index (ESI), seedling emergence speed (SES) plant height and soybean yield in different intervals between cover crop desiccation with glyphosate 480 (3 L ha-1) and BRS 232 cultivar sowing. Plots of 5 x 2.5 m with 1 m of border received four treatments with BO cover crops and four with CO as well as a control for each cover crop, at random, with five replications. The plots were desiccated in intervals of 1, 10, 20 and 30 days before soybean seeding. The harvest was manual while yield was adjusted to 13% of moisture content. The experimental design was completely randomized with splitplots and means compared by the Scott and Knott test at 5% of significance. The results showed that CO of cover crops can be recommended for soybean to obtain a more vigorous seedling emergence, from 10 days after cover crop desiccation.

Soybean growth and yield under cover crops

The use of cover crops in no-tillage systems can provide better conditions for the development of soybean plants with positive effects on grain yield and growth analysis techniques allow researchers to characterize and understand the behavior of soybean plants under different straw covers. Thus, the aim of this study was to characterize, using growth analysis, yield components and agronomic performance of soybean under common bean, Brachiaria brizantha and pearl millet straws. The experiment was performed on a soil under cerrado in the municipality of Santo Antônio de Goiás, GO. The experiment was arranged in a randomized complete block design with three treatments (cover crops) and five replications. Soybean grain yield was lower in the B. brizantha straw treatment (3,708 kg ha-1) than both in the pearl millet (4.772 kg ha-1) and common bean straw treatments (5,200 kg ha-1). The soybean growth analysis in B. brizantha, pearl millet and common bean allowed characterizing the variation in the production of dry matter of leaves, stems, pods and total and leaf area index that provided different grain yields. The cover crop directly affects the soybean grain yield.

Action of auxin inhibitors on growth and grain yield of soybean

Soybean genotypes grown in sub-tropical climate may exhibit lodging. The plant lodging is influenced by soil type and fertility level, sowing date, latitude and altitude of the location, plant population and conditions of crop development. Plant regulators and herbicides are able to avoid or reduce plant lodging. This study aimed to verify the effects of the growth regulators TIBA and daminozide on vegetative growth and yield of soybean cultivar CD 214 RR. The experiment was carried out at a field in randomized block design with four replications in a factorial scheme. The A factor was represented by the combination of regulators TIBA and daminozide and its concentrations, and the Factor B was seven times of evaluation of injury and plant height or eight times of evaluation of lodging. In the range of doses used, the application of daminozide resulted in greater injury to soybean plants than TIBA. The smaller plant height was achieved by the application of 6 g ha-1 of TIBA and 1200 g ha-¹ of daminozide. Treatments with daminozide (100 g ha-¹) and TIBA (10 g ha-1) stood out due to the reduced lodging of soybean plants. Grain weight increased linearly when the levels of TIBA increased. There was a negative correlation between lodging and grain yield and a positive correlation between plant height and lodging. There was also a negative correlation between injury caused by the application of plant regulators and lodging.

Heavy metal toxicity in rice and soybean plants cultivated in contaminated soil

Heavy metals can accumulate in soil and cause phytotoxicity in plants with some specific symptoms. The present study evaluated the specific symptoms on rice and soybeans plants caused by excess of heavy metals in soil. Rice and soybean were grown in pots containing soil with different levels of heavy metals. A completely randomized design was used, with four replications, using two crop species and seven sample soils with different contamination levels. Rice and soybean exhibited different responses to the high concentrations of heavy metals in the soil. Rice plants accumulated higher Cu, Mn, Pb and Zn concentrations and were more sensitive to high concentrations of these elements in the soil, absorbing them more easily compared to the soybean plants. However, high available Zn concentrations in the soil caused phytotoxicity symptoms in rice and soybean, mainly chlorosis and inhibited plant growth. Further, high Zn concentrations in the soil reduced the Fe concentration in the shoots of soybean and rice plants to levels considered deficient.

Urease inhibitor (NBPT) and efficiency of single or split application of urea in wheat crop

NBPT (N-(n-butyl) thiophosphoric triamide), a urease inhibitor, has been reported as one of the most promising compounds to maximize urea nitrogen use in agricultural systems. The objective of this study was to evaluate the performance of irrigated wheat fertilized with urea or urea + NBPT as single or split application. The experiment was conducted from June to October 2006 in Viçosa, MG, Brazil. The experimental design followed a 2×2 factorial scheme, in which urea or urea + NBPT were combined with two modes of application: full dose at sowing (60kg ha-1) or split (20kg ha-1 at sowing + 40kg ha-1 as topdressing at tillering), in randomized blocks with ten replications. The split application of nitrogen fertilization does not improve the yield wheat under used conditions. The use of urease inhibitor improves the grain yield of wheat crop when urea is applied in topdressing at tillering, but its use does not promote difference when urea is applied in the furrow at planting.

Influence of foliar fertilization with manganese on germination, vigor and storage time of RR soybean seeds

ABSTRACTThis study aimed to evaluate the influence of foliar fertilizer doses containing Mn of phenological stages of suggested application in RR soybeans, to recover management damages with glyphosate at postemergence application on seed vigor in post-harvest and post six months storage. The seeds originated from a field experiment conducted , which included two applications of glyphosate, concomitant with foliar fertilizer in growth stages V4 and V6, with 0.00, 113.50 and 227.00 mg ha-1doses of Mn2+. Germination, GSI (Germination Speed Index), electrical conductivity tests and the first count of seeds were conducted. The application of Mn did not affect the physiological quality of RR soy in postharvest. However, in post-storage, higher doses of Mn had a negative effect on tests of abnormal seedlings, GSI and electrical conductivity. The applications of Mn, regardless of the developmental stage, did not interfere in the germination and first count tests, with and without storage. The electrical conductivity test showed a higher correlation with the seed germination test in the post-harvest treatment.

Absorption of primary macronutrients and soybean growth at different compactation densities and moisture levels in a silt loam soil

ABSTRACT Soybean cultivation is increasing rapidly in the region of Alto Vale do Itajaí, State of Santa Catarina, where there is a predominance of silt soils. The objective of this work was to evaluate the content of primary macronutrients in shoots and shoot and root vegetative growth of soybean (Glicine max L. Merrill) grown in a silt-loam soil under different compactation densities and moisture levels. A randomized block design in a 4x4 factorial arrangement was used, with four compactation densities: 1.00; 1.20; 1.40 and 1.60 Mg m-3, and four soil moisture levels: 0.130; 0.160; 0.190 and 0.220 kg kg-1 and four replications. Each pot consisted of the overlapping of three 150-mm PVC rings, where soil was maintained in the higher and lower part of the pot with a density of 1.00 Mg m-3 and in the intermediate ring, the compactation densities were increased. Values of soil density higher than 120 Mg m-3 negatively affected N, P and K uptake by soybean plants, as well as the plant mass of the shoots and roots. The higher levels of soil moisture reduced the compaction effect and promoted better absorption of P and K.