Kinetic measurements during transient film growth on zinc

The electrochemical behaviour of zinc has been extensively studied in alkaline and acid media, but only a few studies have been reported in neutral solutions, particularly in deaerated media. Zinc passivation in neutral medium and the effect of the ClO4- ion on the nucleation and growth of the passive layer is studied in this paper by a transient technique at different electrolyte concentrations and applied potentials. ZnO growth rate was shown to decrease with increasing electrolyte concentration. Moreover, passive layer growth occurred followed by pitting nucleation and growth. Film growth and pit nucleation are explained by means of the Macdonald and Engell-Stolica models.

Semi-selective medium for Fusarium graminearum detection in seed samples

Fungi of the genus Fusarium cause a variety of difficult to control diseases in different crops, including winter cereals and maize. Among the species of this genus Fusarium graminearum deserves attention. The aim of this work was to develop a semi-selective medium to study this fungus. In several experiments, substrates for fungal growth were tested, including fungicides and antibiotics such as iprodiona, nystatin and triadimenol, and the antibacterial agents streptomycin and neomycin sulfate. Five seed samples of wheat, barley, oat, black beans and soybeans for F. graminearum detection by using the media Nash and Snyder agar (NSA), Segalin & Reis agar (SRA) and one-quarter dextrose agar (1/4PDA; potato 50g; dextrose 5g and agar 20g), either unsupplemented or supplemented with various concentrations of the antimicrobial agents cited above. The selected components and concentrations (g.L-1) of the proposed medium, Segalin & Reis agar (SRA-FG), were: iprodiona 0.05; nystatin 0,025; triadimenol 0.015; neomycin sulfate 0.05; and streptomycin sulfate, 0.3 added of ¼ potato sucrose agar. In the isolation from seeds of cited plant species, the sensitivity of this medium was similar to that of NSA but with de advantage of maintaining the colony morphological aspects similar to those observed in potato-dextrose-agar medium.

Influence of Insulin-like Growth Factor I (IGF-I) on the survival and the in vitro development of caprine preantral follicles

The aim of this study was to investigate the effects of the insulin-like growth factor -I (IGF-I) on survival, activation (transition from primordial to primary follicles) and growth of caprine preantral follicles cultured in vitro. Fragments of ovarian cortex were cultured for one and seven days in the absence or presence of IGF-I (0, 50 and 100ng/ml). The non-cultured and cultured tissues were processed and analyzed by histology and transmission electron microscopy. The culture for one day in a medium with 100ng/ml of IGF-I showed 86.7% of morphologically normal follicles. These results were similar (P>0.05) to the percentage of normal follicles found in the control (96.7%). It was also found that this medium increased the percentage of follicular activation (developing follicles) with one day of culture. The oocyte and follicular diameters remained similar to the control by culturing for one day in a medium containing 100ng/ml of IGF-I. The ultrastructural analysis did not confirm the integrity of the follicular fragments in a medium containing IGF-I (100ng/ml) after one and seven days of culture. In conclusion, this study demonstrated that the addition of 100 ng/ml of IGF-I in the culture medium enables the development of preantral follicles of goats with one day of culture. However, it is not sufficient to maintain the follicular integrity and the follicular survival rate after seven days of culture.

Influence of wheat straw and rhizosphere on seed germination, early seedling growth and bio-chemical attributes of Trianthema portulacastrum

Decomposing wheat (Triticum aestivum) straw and rhizosphere-infested soil were evaluated for their suppressive activity against horse purslane (Trianthema portulacastrum), a noxious summer weed in Pakistan. Two separate pot studies were carried out. Wheat straw was incorporated at 4, 6 and 8 g kg-1 soil five days before the sowing of horse purslane. Pots without straw incorporation were maintained as control. In a second study, soil was taken from 15 and 30 cm depths from a previously cropped wheat field immediately after its harvest and was used as growing medium. Soil from an intentionally uncropped area of the same field was used as control. Suppressive activity was measured in terms of germination dynamics, seedling growth, and biochemical attributes such as chlorophyll contents, total soluble phenolics, soluble protein and antioxidant enzymes. Germination, seedling growth, chlorophyll contents and soluble protein of horse purslane were all negatively influenced. Higher phenolics and enhanced activities of antioxidant enzymes were noticed in response to wheat residues incorporation and its rhizosphere soil. Both studies established that the phytotoxic influence of wheat straw and wheat-infested rhizosphere soil on horse purslane can further be exploited for horse purslane management as a sustainable approach.

Effects of iron-ore particles on propagule release, growth and photosynthetic performance of Sargassum vulgare C. Agardh (Phaeophyta, Fucales)

The effect of iron-ore particles on the propagule release and growth of Sargassum vulgare C. Agardh was tested under treatments with different concentrations of iron-ore particles: 0.1, 1.0, 10.0 g.L-1 and a solution of 10.0 g.L-1 of filtered iron-ore. Filtered seawater was used as control. Photosynthesis vs. irradiance (P-I) curves were calculated for S. vulgare in the presence of iron-ore and in seawater. There was no significant difference in the number of propagules released by the receptacles or in the percentage of zygote formation among the treatments. The released propagules acted like aggregation centers for the particles, those more heavily coated with iron (10.0 g.L-1) exhibiting the highest sinking velocity (32.6 ± 9.8 mm.s-1). No difference in the percentage of embryo survival was detected during the first week in culture. After four weeks the embryos grew in all treatments. Maximum frond development (5.3 ± 0.8 mm) was observed in treatment of seawater enriched with Provasoli's medium (PES) while initial filoids did not develop in three treatments without PES and with iron-ore (0.1 g.L-1, 1.0 g.L-1 and 10.0 g.L-1). The values for Pmax, alpha and respiration showed no significant differences between the P-I curves. The calculated value for I K was 106.26 µmol.m-2.s-1 to the control curve and 981.49 µmol.m-2.s-1 to the iron-ore curve. The results indicate that the iron-ore particles in high concentration reduce the growth of S. vulgare as they recovered the embryos, juveniles and young plants. In contrast, the presence of the particles did not affect the release of gametes, percentage of zygote formation or the percentage of embryo survival.

Effect of nitrate and ammonium on the growth and protein concentration of Microcystis viridis Lemmermann (Cyanobacteria)

Cyanobacteria are a very important group in aquatic systems, particularly in eutrophic waters. Therefore studies about their success in the environment are essential. Many hypotheses have tried to explain the dominance of Cyanobacteria, and several emphasized the importance of various nitrogen sources for the success of the group. In this study, we measured the effect of ammonium and nitrate on the growth and protein concentration of Microcystis viridis (Cyanobacteria). This species is well-known because bloom formation in eutrophic waters. The study was carried out, in experimental batch cultures, using the WC medium with different nitrogen sources: ammonium, nitrate, ammonium + nitrate (50% ammonium + 50% nitrate) and ammonium at different concentrations (to test for possible NH4+ toxicity). Protein, ammonium and nitrate concentrations were measured at end of each experiment, whereas samples for cell counts were taken daily. Results showed that Microcystis viridis grew faster with ammonium (µ = 0.393 day-1) than with nitrate (µ = 0.263 day-1) and ammonium + nitrate (µ = 0.325 day-1). This pattern is explained by the metabolism of ammonium that presents higher uptake and assimilation rates than nitrate. Maximum cell concentration, however, was higher in the ammonium + nitrate treatment, followed by nitrate treatment. Higher protein concentration were observed in the treatment with nitrate. In the ammonium toxicity test, no difference between the control and NH4+ at 50% was found. Thus, the ammonium concentrations used in these experiments were not toxic. Our results suggest that Cyanobacteria is able to grow on both nitrogen sources even if ammonium may allow faster growth and bloom development.

Conditioned medium from activated spleen cells supports the survival of rat retinal cells in vitro

Cytokines are a heterogeneous group of molecules that have been associated with several functions in the nervous system, such as survival and differentiation of neuronal and glial cells. In the present study, we demonstrated that conditioned medium from spleen cells activated with concanavalin A increased neuritogenesis and survival of retinal cells, as measured by biochemical and morphological criteria. Our data showed that conditioned medium induced a five-fold increase in the amount of protein after 120 h in vitro. This effect was not inhibited by the blockade of voltage-dependent L-type calcium channels with 5.0 µM nifedipine. However, the use of an intracellular calcium chelator (15.0 µM BAPTA-AM) inhibited this effect. Our results support the idea that factors secreted by activated lymphocytes, such as cytokines, can modulate the maintenance and the differentiation of rat retinal cells in vitro, indicating a possible role of these molecules in the development of retinal cells, as well as in its protection against pathological conditions

Effect of conditioned medium of mesenchymal stem cells on the in vitro maturation and subsequent development of mouse oocyte

Mesenchymal stem cells (MSCs) secrete a variety of cytokines and growth factors in addition to self-renewal and multiple forms of differentiation. Some of these secreted bioactive factors could improve meiotic maturation in vitro and subsequent embryo developmental potential. The aim of the present study was to determine whether in vitro maturation (IVM) of mouse oocyte with or without cumulus cells could be improved by contact with conditioned medium (CM) of MSCs as well as the efficiency of CM to support follicular growth and oocyte maturation in the ovarian organ of mice cultured on soft agar. The developmental potential of matured oocyte was assessed by blastocyst formation after in vitro fertilization (IVF). Germinal vesicle stage oocytes with or without cumulus cells were subjected to IVM in either CM, Dulbecco's modified Eagle's medium (DMEM), α-minimum essential medium (α-MEM) or human tubal fluid (HTF). Approximately 120 oocytes were studied for each medium. CM produced a higher maturation rate (91.2%) than DMEM (54.7%), α-MEM (63.5%) and HTF (27.1%). Moreover, CM improved embryo development to blastocyst stage significantly more than DMEM and HTF (85 vs 7% and 41.7%, respectively) but there was no significant difference compared with α-MEM (85 vs 80.3%). The behavior of cortical granules of IVM oocytes cultured in CM revealed cytoplasmic maturation. Moreover, CM also supported preantral follicles growth well in organotypic culture on soft agar resulting in the maturation of 60% of them to developmentally competent oocytes. The production of estrogen progressively increased approximately 1-fold every other day during organ culture, while a dramatic 10-fold increase in progesterone was observed 17 h after human chorionic gonadotropin stimulus at the end of culture. Thus, CM is an effective medium for preantral follicle growth, oocyte maturation, and sequential embryo development.

Effects of the conditioned medium of mesenchymal stem cells on mouse oocyte activation and development

Mesenchymal stem cells (MSCs) have been reported to secrete a variety of cytokines and growth factors acting as trophic suppliers, but little is known regarding the effects of conditioned medium (CM) of MSCs isolated from femurs and tibias of mouse on the artificial activation of mouse oocytes and on the developmental competence of the parthenotes. In the current study, we investigated the effect of CM on the events of mouse oocyte activation, namely oscillations of cytosolic calcium concentration ([Ca²+]i), meiosis resumption, pronucleus formation, and parthenogenetic development. The surface markers of MSCs were identified with a fluorescence-activated cell sorter. The dynamic changes of the spindle and formation of pronuclei were examined by laser-scanning confocal microscopy. Exposure of cumulus-oocyte complexes to CM for 40 min was optimal for inducing oocyte parthenogenetic activation and evoking [Ca²+]i oscillations similar to those evoked by sperm (95 vs 100%; P > 0.05). Parthenogenetically activated oocytes immediately treated with 7.5 µg/mL cytochalasin B (CB), which inhibited spindle rotation and second polar body extrusion, were mostly diploid (93 vs 6%, P < 0.01) while CB-untreated oocytes were mostly haploid (5 vs 83%, P < 0.01). Consequently, the blastocyst rate was higher in the CB-treated than in the CB-untreated oocytes. There was no significant difference in developmental rate between oocytes activated with CM and 7% ethanol (62 vs 62%, P > 0.05), but the developmental competence of the fertilized oocytes was superior to that of the parthenotes (88 vs 62%, P < 0.05). The present results demonstrate that CM can effectively activate mouse oocytes, as judged by the generation of [Ca²+]i oscillations, completion of meiosis and parthenogenetic development.

Interaction between growth differentiation factor 9, insulin-like growth factor I and growth hormone on the in vitro development and survival of goat preantral follicles

The objective of this study was to determine the effects of GDF-9, IGF-I, and GH alone or combined on preantral follicle survival, activation and development after 1 and 7 days of in vitro culture. Either fresh (non-cultured) or cultured ovarian tissue was processed for histological and fluorescence analysis. For all media tested, the percent of normal follicles was greater when compared to minimum essential medium supplemented (MEM+) alone, except when ovarian tissue was cultured with GDF-9/IGF-I or GDF-9/GH (P < 0.05). Fluorescence analysis showed that the percent of viable follicles after 7 days of culture was similar for non-cultured tissue and for all treatments tested. The percent of primordial follicles was reduced (P < 0.05) and there was a significant and concomitant increase in the percent of intermediate and primary follicles in all treatments tested after 7 days of culture when compared to non-cultured tissue. After 7 days of culture, the highest percent of intermediate follicles was observed with IGF-I/GH (61.3%), and the highest percent of primary follicles was achieved with IGF-I (57.7%). After 7 days of culture in MEM+ containing GDF-9, IGF-I and GH alone or in all associations, a significant increase in follicular diameter was observed when compared to MEM+ alone and non-cultured tissue. In conclusion, GDF-9, IGF-I and GH alone or in combination maintain preantral follicle survival and promote primordial follicle activation. Nevertheless, the data showed that IGF-I/GH and IGF-I alone are efficient in promoting the transition from primordial to intermediate follicles and from intermediate to primary follicles, respectively.

Fusarium graminearum growth inhibition mechanism using phenolic compounds from Spirulina sp

The application of natural antifungal substances is motivated by the need for alternatives to existing methods that are not always applicable, efficient, or that do not pose risk to consumers or the environment. Furthermore, studies on the behaviour of toxigenic species in the presence of natural fungicides have enabled their safe application in the food chain In this study, Spirulina LEB-18 phenolic extract was assessed for its antifungal activity on 12 toxigenic strains of Fusarium graminearum isolated from barley and wheat. The susceptible metabolic pathways were assessed through the determination of structural compounds (glucosamine and ergosterol) and enzyme activity of the microorganisms' primary metabolism. The results indicate that phenolic extracts reduced the growth rate of the toxigenic species investigated. The IC50 was obtained by applying 3 to 8% (p/p) of phenolic compounds in relation to the culture medium. The use of this natural fungicide proved promising for the inhibition of fungal multiplication, especially in terms of the inactivation of enzymatic systems (amylase and protease) of Fusarium graminearum.

Ability of a Lactobacillus rhamnosus strain cultured in milk whey based medium to bind aflatoxin B1

This study aimed to compare Lactobacillus rhamnosus growth in MRS (de Man, Rogosa and Sharpe) broth and a culture medium containing milk whey (MMW) and to evaluate aflatoxin B1 (AFB1) adsorption capacity by bacterial cells produced in both culture media. L. rhamnosus cells were cultivated in MRS broth and MMW (37 °C, 24 hours), and bacterial cell concentration was determined spectrophotometrically at 600 nm. AFB1 (1 µg/ml) adsorption assays were conducted using 1 x 10(10) non-viable L. rhamnosus cells (121 °C, 15 minutes) at pHs 3.0 and 6.0 and contact time of 60 minutes. AFB1 quantification was performed by High Performance Liquid Chromatography. Bacterial cell concentration in MMW was higher (9.84 log CFU/ml) than that in MRS broth (9.63 log CFU/ml). There were no significant differences between AFB1 binding results at the same pH value (3.0 or 6.0) for the cells cultivated in MRS broth (46.0% and 35.8%, respectively) and in MMW (43.7% and 25.8%, respectively), showing that MMW can adequately replace the MRS broth. Therefore, it can be concluded that the use of L. rhamnosus cells cultivated in MMW offers advantages such as reduction in large scale production costs, improvement of environmental sustainability, and being a practicable alternative for decontamination of food products susceptible to aflatoxin contamination.

Production of medium-chain-length polyhydroxyalkanoate by Pseudomonas oleovorans grown in sugary cassava extract supplemented with andiroba oil

Pseudomonas oleovorans were grown on sugary cassava extracts supplemented with andiroba oil for the synthesis of a mediumchain- length polyhydroxyalkanoate (PHA MCL). The concentration of total sugars in the extract was approximately: 40 g/L in culture 1, 15 g/L in cultures 2 and 3, and 10 g/L in culture 4. Supplementation with 1% andiroba oil and 0.2 g/L of (NH4)2HPO4 was performed 6.5 hours after growth in culture 3, and supplementation with the same amount of andiroba oil and 2.4 g/L of (NH4)2HPO4 was performed at the beginning of growth in culture 4. The synthesis resulted mainly in 3-hydroxy-decanoate and 3-hydroxy-dodecanoate units; 3-hydroxy-butyrate, 3-hydroxy-hexanoate; and 3-hydroxy-octanoate monomers were also produced but in smaller proportions. P. oleovorans significantly accumulated PHA MCL in the deceleration phase of growth with an oxygen limitation but with sufficient nitrogen concentration to maintain cell growth. The sugary cassava extract supplemented with andiroba oil proved to be a potential substrate for PHA MCL production.

Slow-release and organic fertilizers on early growth of Rangpur lime

Slow-release and organic fertilizers are promising alternatives to conventional fertilizers, as both reduce losses by leaching, volatilization and problems of toxicity and/or salinity to plants. The objective of this work was to evaluate the effect of different rates of the organic fertilizer Humato-Macota® compared with the slow-release fertilizer Osmocote® on the growth and nitrogen content in the dry matter of Rangpur lime. A field experiment was conducted in a factorial completely randomized design with an additional treatment (4 x 4 +1). The first factor consisted of four Humato­Macota® rates (0, 1, 2, and 3%) applied to the substrate; the second factor consisted of the same Humato-Macota® concentrations, but applied as fortnightly foliar sprays; the additional treatment consisted of application of 5 kgm-3 Osmocote® 18-05-09. Means of all growth characteristics (plant height, total dry matter, root/shoot ratio and leaf area) and the potential quantum yield of photosystem II (Fv/Fm) were higher when plants were fertilized with the slow-release fertilizer. The organic fertilizer applied alone did not meet the N requirement of Rangpur lime.

Root growth of Arabidopsis thaliana (L.) Heynh. treated with humic acids isolated from typical soils of Rio de Janeiro state, Brazil

Humic substances isolated from soil organic matter had been used as stimulators of plant metabolism. Arabidopsis thaliana (L.) Heynh. with only five chromosomes, short cycle and size, is an important model to evaluate the physiological effects of these substances, which are qualitatively and quantitatively influenced by morphogenesis, mineralogy and chemistry of soils. The objective of this study was to evaluate the ambience effects on bioactivity of humic acids. A and B horizons of four typical soils of the North Fluminense were sampled. After isolation and purification, humic acids were applied to plants in increasing concentrations. The number and length of lateral roots and main root length were evaluated and, subsequently, the concentrations of maximum stimulation were determined by dose-response curves and regression equations. The results showed that more stable humic acids isolated from soil in less advanced stages of weathering, high activity clay and high base saturation resulted in better physiological stimulants for Arabidopsis.