Fermentation as a bio-process to obtain functional soybean flours

The effect of fermentation on the antioxidant compounds [vitamins C and E, total phenolic compounds (TPC), and reduced glutathione (GSH)], and antioxidant capacity [superoxide anion scavenging activity (SOD-like activity), peroxyl radical-trapping capacity (PRTC), inhibition of phosphatidylcholine (PC) peroxidation, and Trolox equivalent antioxidant capacity (TEAC)] of soybean (Glycine max cv. Merit) was studied. Fermentation was carried out in solid state in cracked seeds inoculated with Aspergillus oryzae, Rhizopus oryzae, Bacillus subtilis, and Lactobacillus plantarum and in liquid state either in cracked seeds or milled soybean flours fermented naturally by only the microorganisms present in the seeds or by inoculation with L. plantarum. Vitamin C was not detected in the studied samples. Fermentation caused a decrease in vitamin E activity, except when cracked seed was fermented with A. oryzae, R. oryzae, or B. subtilis that increased 31, 30, and 89%, respectively. Fermentation produced an increase in TPC content and did not affect or reduce the GSH content. Fermentation decreased SOD-like activity drastically, while PRTC increased except when it was carried out naturally in cracked seed. TEAC values rose sharply when soybeans were fermented with B. subtilis. Processed soybean extracts inhibited PC peroxidation in comparison with the control assay. On the basis of the results obtained, the relative contributions of vitamin E, TPC, and GSH to antioxidant capacity were calculated and results showed a very high TPC contribution and a low contribution of GSH and vitamin E activity. Optimum results for functional soybean flours were achieved when fermentation was carried out with B. subtilis inoculum.     

Diurnal and seasonal water variations of temperature,pH, redox potential and conductivity in gnammas (weathering pits): Implications for chemical weathering

A gnamma or weathering pit is a small-sized cavity sculpted into solid rock which refills with rainwater. This study describes the main hydrological processes in gnammas and the underlying chemical interactions with the environment. Four gnammas developed in a granite tor from central Spain were chosen for monitoring and studying the dynamics of the gnamma–water system. Parameters monitored were air and water temperature, pH, ORP (redox potential), and conductivity. These measurements describe the system dynamics, without the need for special sampling or other potential interferences under low water volumes. Additionally, chemical analyses were carried out to contrast the chemical properties with the chemical composition. As a consequence of water volumes less than 100 L, daily fluctuations tended to cause the largest amplitude changes. Photosynthesis and respiration/putrefaction processes in the gnamma are responsible for daily pH fluctuations up to 4 units, with pH values often greater than 9. ORP generally alternates from positive to negative values as the solution oscillates diurnally between basic and acidic conditions. Conductivity values ∼ 10 μS/cm occur most of the year, with only minor daily oscillations. During dry periods, conductivity increases significantly with maximum values > 400 μS/cm. These high conductivity values reflect atmospheric inputs by summer rainstorms after prolonged dry periods. Therefore, conductivity does not relate directly with chemical species dissolved from the host rock. Silica was the main weathering product of granite dissolution during the year, with highest concentrations (> 200 ppm) in the rainy season. Low Si values measured during the dry season are in accordance with observed precipitation of amorphous silica within the gnamma. Seasonal variations in concentration are also present in Al and Fe as a response to ORP variations and evaporative dynamics, causing mineral dissolution and new mineral precipitation during the year. Relations among the measured parameters are influenced by several factors, including biological processes, temperature effects and water level variations. Factors such as large pH oscillations, basic pH values, acids from biological activity, and alkali cations all increase silicate solubility, thus favoring granite dissolution. Overflow of water during the winter, when the water is chemically enriched, is an effective mechanism to remove weathering products.     

Nitrogen Release from Slow-Release Fertilizers in Soils with Different Microbial Activities

Soil microbial activity is recognized as an important factor affecting nitrogen (N) release from slow-release fertilizers. However,studies on the effect of size and activity of soil microflora on fertilizer degradation have provided contrasting results. To date, no clear relationships exist between soil microbial activity and the release of N from slow-release fertilizers. Hence, the aim of this study was to better understand such relationships by determining the release of N from three slow-release fertilizers in soils with different microbial activities. Soils were amended with urea-formaldehyde (UF), isobutylidene diurea (IBDU), and crotonylidene diurea (CDU). Urea, a soluble fertilizer, was used as the control. Fertilized soil samples were placed in a leaching system, and the release of N was determined by measuring ammonium-N and nitrate-N concentrations in leachates during 90 d of incubation. Non-linear regression was used to fit N leaching rate to a first-order model. In all the treated soils, N was released in the order: urea (89%–100%) IBDU (59%–94%) UF (46%–73%) CDU (44%–56%). At the end of incubation, N released from CDU did not differ (P 0.05) among soils. On the contrary, UF and IBDU released significantly lower (P 0.05) amounts of N in the soil with higher microbial activity and lower pH.The rate constant (K_0) for UF was lower (P 0.05) in the soil with lower pH. Taken together, our results indicated that soil microbial size and microbial activity had a marginal effect on fertilizer mineralization.     

Interactive effect between Cu‐adapted arbuscular mycorrhizal fungi and biotreated agrowaste residue to improve the nutritional status of Oenothera picensis growing in Cu‐polluted soils

The interactive effect of sugar beet (SB) agrowaste and arbuscular mycorrhizal (AM) fungi inoculation in response to increasing Cu levels was evaluated in the metallophyte Oenothera picensis. Plants were grown in a Cu‐added soil (0, 100, or 500 mg Cu kg^?1), in presence or absence of SB, and inoculated with: (1) indigenous Cu adapted mycorrhiza (IM) isolated from Cu‐polluted soils; (2) Claroideoglomus claroideum (CC); or (3) maintained uninoculated (control). Sugar beet application produced an increase in shoot biomass of 2 to 7 times, improving plant nutritional status and allowing their survival at the highest Cu concentrations. Moreover, AM fungi utilization had a positive effect promoting the plant establishment; nevertheless, Cu plant concentration as well as the mycorrhizal development in terms of AM colonization, AM spore density, and glomalin production were strictly dependent of the AM fungi strains used. Remarkable differences between AM fungi strains were observed at the highest soil Cu level where only plants colonized by IM were able to survive and grow when no SB residue was added. An interactive effect between AM fungi and SB produced a higher plant growth than plants without the amendment application, improving the plant establishment and allowing their survival at highest copper concentrations, suggesting that this combination could be used as a biotechnological tool for the phytoremediation of Cu‐polluted soils.     

Quantitative changes in the normal and apoptotic thymocytes of pigs treated with anabolic doses of the beta2 adrenergic agonist clenbuterol

Although the beta2 adrenergic agonists have been seen to have important effects on the mechanisms regulating the development and death of T-cells in the thymus, the side-effects on the immune system of anabolic treatments of these substances have hardly been considered. In order to evaluate the effects exerted by the beta2 adrenergic agonist clenbuterol on the thymocyte population, the thymus of eight pigs treated with anabolic doses of this substance was studied by morphometric methods, regarding apoptotic (terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling (TUNEL)-positive) and normal (TUNEL-negative) cells. The thymus of another eight pigs fed without clenbuterol served as a control. The clenbuterol treatment had a clear effect on the thymocyte size, decreasing their mean nuclear area. The T-cell apoptosis index was also affected by the clenbuterol, significantly increasing the apoptosis percentage in the treated group with respect to the control. In the light of our results, the clenbuterol induced thymocyte apoptosis throughout the thymus and caused morphometric changes in the thymocyte population, which was in line with the immunosuppressive role attributed to other beta2 adrenergic agonists.     

Multiple generalized follicular cysts in a stallion

This case report describes a case of multiple follicular cysts in a 4-year-old Spanish purebred stallion. The lesions ranged in size from 0.5 to 3 cm in diameter, and were firm, well circumscribed and nonpruritic. They developed over a 2-year period with a generalized distribution affecting all body regions. Five nodules were removed and histopathologically corresponded to simple epidermal cysts (infundibular and isthmus-catagen) with squamous epithelium and a keratin filled cavity. Lesions were not evident at birth but their number, early age of detection, slow growth and lack of previous trauma suggested that they were congenital. To the authors' knowledge, this condition has not previously been reported either in young horses or in Spanish purebred horses.     

Novel Prunus rootstock somaclonal variants with divergent ability to tolerate waterlogging

Plants require access to free water for nutrient uptake, but excess water surrounding the roots can be injurious or even lethal because it blocks the transfer of free oxygen between the soil and the atmosphere. Genetic improvement efforts in this study were focused on the increased tolerance in roots to waterlogging. Among a pool of clones generated in vitro from leaf explants of rootstock Mr.S.2/5 of Prunus cerasifera L., the S.4 clone was flood tolerant whereas the S.1 clone was sensitive. The S.4 clone formed adventitious roots on exposure to flooding. Moreover, the chlorophyll content and mitochondrial activity in the leaf and root, soluble sugar content, alcohol dehydrogenase activity and ethylene content were different between the clones. The sorbitol transporter gene (SOT1) was up-regulated during hypoxia, the alcohol dehydrogenase genes (ADH1 and ADH3) were up-regulated in the leaves and down-regulated in the roots of the S.4 clone during hypoxia, and the 1-aminocyclopropane-1-oxidase gene (ACO1) was up-regulated in the leaves and roots of the S.4 clone during hypoxia and down-regulated in the wild-type roots. In addition, in the S.4 root, hypoxia induced significant down-regulation of a glycosyltransferase-like gene (GTL), which has a yet-undefined role. Although the relevant variation in the S.4 genome has yet to be determined, genetic alteration clearly conferred a flooding-tolerant phenotype. The isolation of novel somaclonals with the same genomic background but with divergent tolerance to flooding may offer new insights in the elucidation of the genetic machinery of resistance to flooding and aid in the selection of new Prunus rootstocks to be used in various adverse environments.     

Strecker degradation of phenylalanine initiated by 2,4-decadienal or methyl 13-oxooctadeca-9,11-dienoate in model systems

The reaction of 2,4-decadienal and methyl 13-oxooctadeca-9,11-dienoate with phenylalanine was studied to determine if alkadienals and ketodienes are able to produce the Strecker-type degradation of amino acids to the corresponding Strecker aldehydes. When reactions were carried out at 180 degrees C, both carbonyl compounds degraded phenylalanine to phenylacetaldehyde, among other compounds. The yield of the phenylacetaldehyde produced depended on the reaction pH and increased linearly with both the amount of the lipid and the reaction time. The yield of this conversion was approximately 8% when starting from decadienal and approximately 6% when starting from methyl 13-oxooctadeca-9,11-dienoate, and the reaction rate was lower for the ketone than for the aldehyde. Simultaneous to these reactions, the lipid was converted into pyrrole, pyridine, or aldehyde derivatives as a result of several competitive reactions. In particular, 9-14% of the decadienal was converted into hexanal under the assayed conditions. All these reactions are suggested to be produced as a consequence of the oxidation of the alkadienal or the ketodiene to the corresponding epoxyalkenal or unsaturated epoxyketone, which were identified in the reaction mixtures by GC-MS. All these results suggest that alkadienals and ketodienes, which are quantitatively important secondary lipid oxidation products, can degrade amino acids to their corresponding Strecker aldehydes. Therefore, under appropriate conditions, these products are not final products of the lipid oxidation and can participate in carbonyl-amine reactions analogously to other lipid oxidation products with two oxygenated functions.     

Quantitative structure-antifungal activity relationships of some benzohydrazides against Botrytis cinerea

Fourteen benzohydrazides have been synthesized and evaluated for their in vitro antifungal activity against the phytopathogenic fungus Botrytis cinerea. The best antifungal activity was observed for the N',N'-dibenzylbenzohydrazides 3b-d and for the N-aminoisoindoline-derived benzohydrazide 5. A quantitative structure-activity relationship (QSAR) study has been developed using a topological substructural molecular design (TOPS-MODE) approach to interpret the antifungal activity of these synthetic compounds. The model described 98.3% of the experimental variance, with a standard deviation of 4.02. The influence of an ortho substituent on the conformation of the benzohydrazides was investigated by X-ray crystallography and supported by QSAR study. Several aspects of the structure-activity relationships are discussed in terms of the contribution of different bonds to the antifungal activity, thereby making the relationships between structure and biological activity more transparent.