Interactions between different media and follicle‐stimulating hormone supplementation on in vitro culture of preantral follicles enclosed in ovarian tissue derived from collared peccaries (Pecari tajacu Linneaus, 1758)

The aim was to verify the effect of follicle‐stimulating hormone (FSH) supplementation to α‐MEM+ or TCM199+ media on the in vitro development of ovarian preantral follicles (PFs) derived from collared peccaries. Ovaries (n = 5 pairs) were collected and divided into fragments destined to control group (non‐cultured) or treatments that were cultured for 7 days. The PFs morphology, growth and activation were evaluated by classical histology. The immunohistochemistry markers Ag‐NOR and PCNA were used for nuclear proliferation analysis, and the picrosirius red labelling was used for ovarian extracellular matrix (ECM) evaluation. After 7‐day culture, only the TCM199+ treatment maintained the proportion of intact PFs similar to day 1 (63.2%), but no differences were found among treatments (p > .05). In addition, a significant increase in the growing follicles proportion was verified for all the treatments, indicating follicular activation (p > .05). By the Ag‐NOR analysis, only the TCM199+/FSH maintained the nuclear proliferation similar to the first day (p > .05). The picrosirius red staining revealed that the ECM remained intact in all the treatments (p > .05). We suggest the use of TCM199+ medium supplemented of FSH for the in vitro development of peccaries PFs under 7‐day culturing conditions.     

Antioxidant and Antimicrobial Potential of the Bifurcaria bifurcata Epiphytic Bacteria

Surface-associated marine bacteria are an interesting source of new secondary metabolites. The aim of this study was the isolation and identification of epiphytic bacteria from the marine brown alga, Bifurcaria bifurcata, and the evaluation of the antioxidant and antimicrobial activity of bacteria extracts. The identification of epiphytic bacteria was determined by 16S rRNA gene sequencing. Bacteria extracts were obtained with methanol and dichloromethane (1:1) extraction. The antioxidant activity of extracts was performed by quantification of total phenolic content (TPC), 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity and oxygen radical absorbance capacity (ORAC). Antimicrobial activities were evaluated against Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis, Salmonella enteritidis, Staphylococcus aureus, Saccharomyces cerevisiae and Candida albicans. A total of 39 Bifurcaria bifurcata-associated bacteria were isolated and 33 were identified as Vibrio sp. (48.72%), Alteromonas sp. (12.82%), Shewanella sp. (12.26%), Serratia sp. (2.56%), Citricoccus sp. (2.56%), Cellulophaga sp. (2.56%), Ruegeria sp. (2.56%) and Staphylococcus sp. (2.56%). Six (15.38%) of the 39 bacteria Bifurcaria bifurcata-associated bacteria presented less than a 90% Basic Local Alignment Search Tool (BLAST) match, and some of those could be new. The highest antioxidant activity and antimicrobial activity (against B. subtilis) was exhibited by strain 16 (Shewanella sp.). Several strains also presented high antimicrobial activity against S. aureus, mainly belonging to Alteromonas sp. and Vibrio sp. There were no positive results against fungi and Gram-negative bacteria. Bifurcaria bifurcata epiphytic bacteria were revealed to be excellent sources of natural antioxidant and antimicrobial compounds.     

An Effect of Various Nitrogen Forms Used as Fertilizer on Lupinus albus L. Yield and Protein, Alkaloid and α-Galactosides Content

The influence of various forms of nitrogen supplied to the ground as N₂, NH, NO, [NH + NO], –NH₂, and Nd (N‐deficiency) on Lupinus albus L. cvs. Butan (sweet) and Bac (bitter) yield and protein, alkaloid and α‐galactoside biosynthesis in their seeds were studied. The experiments were performed in a greenhouse on perlite using, in all cases, the same dose of P, K, Mg and micronutrients (B, Zn, Mn, Cu, Mo, Fe). The Nd treatment contained only macro‐ and micronutrients without any nitrogen support. We found that different nitrogen forms used as a fertilizer had a significant influence on the development and yield structure of both lupin cultivars. It was found that the plants developed most favourably when nitrogen was supplied as N₂ and [NH + NO]. For the other forms, the anomalies like necrosis, chlorosis, leaves with small surface of assimilation were noticed. However, these observations were not always reflected by all parameters of yield structure. The forms of nitrogen had a pivotal influence on yield of generative and vegetative parts of lupin. Moreover, significant differences in protein, alkaloid and α‐galactoside content in seeds were observed.     

Purification and characterization of a phytate-degrading enzyme from germinated faba beans (Vicia faba Var. Alameda)

A phytate-degrading enzyme was purified approximately 2190-fold from germinated 4-day-old faba bean seedlings to apparent homogeneity with a recovery of 6% referred to the phytase activity in the crude extract. It behaves as a monomeric protein of a molecular mass of approximately 65 kDa. The phytate-degrading enzyme belongs to the acidic phytases. It exhibits a single pH optimum at 5.0. Optimal temperature for the degradation of sodium phytate is 50 degrees C. Kinetic parameters for the hydrolysis of sodium phytate are K(M) = 148 micromol L(-1) and k(cat) = 704 s(-1) at 35 degrees C and pH 5.0. The faba bean phytase exhibits a broad affinity for various phosphorylated compounds and hydrolyzes phytate in a stepwise manner. The first hydrolysis product was identified as D/L-myo-inositol(1,2,3,4,5)pentakisphosphate.     

Functional properties of purified vicilins from cowpea (Vigna unguiculata) and pea (Pisum sativum) and cowpea protein isolate

The major storage globulins (vicilins) of cowpea (Vigna unguiculata) and pea (Pisum sativum) seeds were purified by ammonium sulfate precipitation, and a semipurified cowpea protein isolate (CPI) was prepared by isoelectric precipitation. Some of the functional properties of these proteins, including solubility, foaming, and emulsifying capacities, were investigated and compared. The solubility of purified cowpea vicilin was reduced at pH 5.0, increasing markedly below and above this value. Pea vicilin exhibited poor solubility between pH 5.0 and pH 6.0, and CPI was little soluble in the pH range from 4.0 to 6.0. At neutral pH, the emulsifying activity indexes (EAI) of purified pea vicilin and CPI were 194 and 291 m(2)/g, respectively, which compare quite favorably to EAIs of 110 and 133 m(2)/g for casein and albumin, respectively. Remarkably, purified cowpea vicilin exhibited an EAI of 490 m(2)/g, indicating a very high emulsifying activity. Purified cowpea and pea vicilins exhibited lower foaming capacities and foam stablity indexes (FSI) than CPI. FSI values of 80 and 260 min were obtained for purified pea and cowpea vicilin, respectively, whereas a FSI value of 380 min was obtained for CPI. These results are discussed in terms of the possible utilization of purified vicilins or protein isolates from pea and cowpea in the food processing industry.     

Antitumor and Antimicrobial Potential of Bromoditerpenes Isolated from the Red Alga,Sphaerococcus coronopifolius

Cancer and infectious diseases continue to be a major public health problem, and new drugs are necessary. As marine organisms are well known to provide a wide range of original compounds, the aim of this study was to investigate the bioactivity of the main constituents of the cosmopolitan red alga, Sphaerococcus coronopifolius. The structure of several bromoditerpenes was determined by extensive spectroscopic analysis and comparison with literature data. Five molecules were isolated and characterized which include a new brominated diterpene belonging to the rare dactylomelane family and named sphaerodactylomelol (1), along with four already known sphaerane bromoditerpenes (2–5). Antitumor activity was assessed by cytotoxicity and anti-proliferative assays on an in vitro model of human hepatocellular carcinoma (HepG-2 cells). Antimicrobial activity was evaluated against four pathogenic microorganisms: Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus and Candida albicans. Compound 4 exhibited the highest antimicrobial activity against S. aureus (IC₅₀ 6.35 µM) and compound 5 the highest anti-proliferative activity on HepG-2 cells (IC₅₀ 42.9 µM). The new diterpene, sphaerodactylomelol (1), induced inhibition of cell proliferation (IC₅₀ 280 µM) and cytotoxicity (IC₅₀ 720 µM) on HepG-2 cells and showed antimicrobial activity against S. aureus (IC₅₀ 96.3 µM).     

Antiproliferative Activity of Fucan Nanogel

Sulfated fucans comprise families of polydisperse natural polysaccharides based on sulfated L-fucose. Our aim was to investigate whether fucan nanogel induces cell-specific responses. To that end, a non toxic fucan extracted from Spatoglossum schröederi was chemically modified by grafting hexadecylamine to the polymer hydrophilic backbone. The resulting modified material (SNFuc) formed nanosized particles. The degree of substitution with hydrophobic chains was close to 100%, as estimated by elemental analysis. SNFfuc in aqueous media had a mean diameter of 123 nm and zeta potential of −38.3 ± 0.74 mV, as measured by dynamic light scattering. Nanoparticles conserved their size for up to 70 days. SNFuc cytotoxicity was determined using the MTT assay after culturing different cell lines for 24 h. Tumor-cell (HepG2, 786, H-S5) proliferation was inhibited by 2.0%–43.7% at nanogel concentrations of 0.05–0.5 mg/mL and rabbit aorta endothelial cells (RAEC) non-tumor cell line proliferation displayed inhibition of 8.0%–22.0%. On the other hand, nanogel improved Chinese hamster ovary (CHO) and monocyte macrophage cell (RAW) non-tumor cell line proliferation in the same concentration range. The antiproliferative effect against tumor cells was also confirmed using the BrdU test. Flow cytometric analysis revealed that the fucan nanogel inhibited 786 cell proliferation through caspase and caspase-independent mechanisms. In addition, SNFuc blocks 786 cell passages in the S and G2-M phases of the cell cycle.     

Hydrogen peroxide,iodine solution and methylene solution highly enhance the hatching rate of freshwater ornamental fish species

The aim of this study was to evaluate the effect of hydrogen peroxide, iodine solution (PVP) and methylene blue on eggs disinfection of three ornamental fish species, Danio rerio, Pterophyllum scalare and Gymnocorymbus ternetzi. The main idea was to create conditions to enhance the hatching rates. Eggs of each species were exposed to different concentrations of hydrogen peroxide (5, 10, 15 and 25 mg/L), PVP (0.25, 0.5, 0.75 and 1 mg/L) and methylene blue (0.5 1, 2 and 3 mg/L). The optimal doses ranged between species and chemicals: for G. ternetzi, the concentrations that high enhanced the hatching rate were 1 mg/L for the PVP treatment, 25 mg/L for the hydrogen peroxide treatment and 3 mg/L for methylene blue treatment; for P. scalare, the best results were achieved with 25 mg/L for hydrogen peroxide treatment and 3 mg/L for methylene blue treatment. By contrast, for all the different chemical did not increased the D. rerio hatching rate. Results showed that hydrogen peroxide and methylene blue are the most versatile, effective and safe to use in these species. On the other hand, PVP can be used but with many precautions due to very low safety margin. Results clearly show that the optimal concentration of chemicals for eggs disinfection is fish species dependent and it is completely wrong to extrapolate concentrations between different chemicals and fish species. Our study suggests that P. scalare can be used as a model in study of effectiveness of new chemicals with potential to disinfect water and increase hatching rates.     

The effects of arsenic on the growth and nutritional status of Anadenanthera peregrina,a Brazilian savanna tree

Anadenanthera peregrina is a Brazilian savanna tree species that occurs naturally in arsenic (As)‐contaminated areas, and its As resistance has been associated with arbuscular mycorrhizal–fungi (AMF) symbiosis. A plant's ability to survive in stressful environments is correlated with its nutrition status, which can be affected by As uptake. The present study evaluated the influence of As on the concentrations and distribution of nutrients in the roots and shoots of A. peregrina grown in the absence of AMF. These plants were grown in substrates spiked with 0, 10, 50, and 100 mg As kg^–1 for 25 d under greenhouse conditions, and the concentrations of essential macro‐ (P, K, Ca, Mg, N, and S) and micro‐ (Fe, Mn, Cu, Zn, B, and Mo) nutrients in the roots and shoots were then determined. Enhanced As levels increased the concentrations of P, S, and N and decreased Ca, Mg, and Fe. Although the deleterious effects of As on the plants were striking, the internal As levels were high, which indicated some tissue tolerance of A. peregrina.