In vitro development of IVF-derived bovine embryos following cytoplasmic microinjection for the episomal expression of the IGF2 gene

Important genomic imprinting changes usually occur following the in vitro production (IVP) of bovine embryos, especially in the imprinting pattern of components of the IGF system. This study aimed to evaluate the effects of a transient episomal overexpression of the IGF2 gene in bovine IVP embryos following embryo cytoplasmic microinjection (CMI) at the 1-cell stage on embryo survival, early and late developmental kinetics and morphological quality up to Day 7 of development. Selected cumulus–oocyte complexes (COCs) were matured and fertilized in vitro and subsequently segregated into six experimental groups: non-CMI control group and five CMI groups at increasing doses (0, 10, 20, 40 and 80 ng/μl) of a GFP vector built for the episomal expression of bovine IGF2. Zygote CMI was effective in delivering the expression vector into the ooplasm, irrespective of the groups, with 58% of positive GFP fluorescence in Day 7 blastocysts. Considering developmental rates and late embryo kinetics, the 10-ng/μl CMI vector dose promoted a lower blastocyst rate (10.4%), but for blastocysts at more advanced stages of development (93.0% blastocysts and expanded blastocysts), and higher number of cells (116.0 ± 3.0) than non-CMI controls (23.3%, 75.0% and 75.0 ± 6.8 were obtained, respectively). In conclusion, CMI at the 1-cell stage did not compromise subsequent in vitro development of surviving embryos, with the 10-ng/μl group demonstrating a possible growth-promoting effect of the IGF2 gene on embryo development, from the 1-cell to the blastocyst stage.     

Near-Infrared Spectroscopy Coupled with Chemometrics Tools: A Rapid and Non-Destructive Alternative on Soil Evaluation

Near-infrared (NIR) spectroscopy is a rapid, non-destructive and accurate technique for analyzing a wide variety of samples, thus, the growing interest of using this technique in soil science. The objective of this study was to evaluate the potential of NIR spectroscopy to predict organic carbon (OC), total nitrogen (TN), available phosphorus (P) and available potassium (K) in the soil. NIR spectra from 20 cm³ of soil samples were acquired on the range of 750 to 2500 nm in diffuse reflectance mode, resolution of 16 cm^?1 and 64 scans. Eight models of calibration/validation were constructed. Calibration and validation models showed that the predictive potential of NIR varied with the specific soil property (OC, TN, P and K) under evaluation and according to the methodology employed in the model construction (cross-validation or test set). Good prediction models were obtained for OC and TN content based on the statistical parameters. Test set methodology was able to predict soil OC, TN, P, and K better than cross-validation methodology.     

Biofortification of soybean grains with foliar application of Li sources

Abstract

Biofortification of soybean grains with lithium (Li) is a strategy to improve a food with high social acceptance, in order to promote health benefits. The aim of this work was to evaluate the development of plants and the production of biofortified soybeans with two Li sources. The experiment was conducted in a completely randomized design with five replicates. Ten treatments were obtained in a 2x5 factorial scheme, comprising two sources of Li (LiOH - Li hydroxide and Li₂SO₄ - Li sulfate) and five doses (0, 30, 60, 90 and 120?mg kg^?1) in stages V4 and R1. The supply of Li promoted significant effects (p?≤?0.05) on the morphology, yield components and nutritional status of soybean plants. The highest grain yield was obtained with the use of Li₂SO₄ at the estimated dose of 45.7?mg kg^?1. The plants accumulated Li between 8.00 and 11.20?mg kg ^?1, respectively, with the use of Li₂SO₄ and LiOH. In the grains, the highest concentrations of Li were obtained with the application of 120?mg kg^?1. Li₂SO₄ and LiOH are good sources for biofortification of soybean grains.     

<Emphasis Type="Italic">Achromobacter insolitus</Emphasis> and <Emphasis Type="Italic">Zoogloea ramigera</Emphasis> associated with wheat plants (<Emphasis Type="Italic">Triticum aestivum</Emphasis>)

This study reports for the first time the presence of diazotrophic bacteria belonging to the genera Achromobacter and Zoogloea associated with wheat plants. These bacterial strains were identified by the analysis of 16S rDNA sequences. The bacterium IAC-AT-8 was identified as Azospirillum brasiliense, whereas isolates IAC-HT-11 and IAC-HT-12 were identified as Achromobacter insolitus and Zoogloea ramigera, respectively. A greenhouse experiment involving a non-sterilized soil was carried out with the aim to study the endophytic feature of these strains. After 40 days from inoculation, all the strains were in the inner of roots, but they were not detected in soil. In order to assess the location inside wheat plants, an experiment was conducted under axenic conditions. Fifteen days after inoculation, preparations of inoculated plants were observed by the scanning electron microscope, using the cryofracture technique, and by the transmission electron microscope. It was observed that all isolates were present on the external part of the roots and in the inner part at the elongation region, in cortex cells, but not in the endodermis or in the vascular bundle region. No colonizing bacterial cells were observed in wheat leaves.     

Response of stevia to nitrogen fertilization and harvesting regime in northeastern Portugal

The adaptation of stevia to the growing conditions of NE Portugal is assessed, including the tolerance of this species to cold temperatures, and the potential to produce biomass when grown as an annual crop and when subjected to various nitrogen (N) rates and two harvesting regimes. Almost all the plants died during the winter of 2014 (minimum temperatures peaked at ?8.0°C), making it necessary to replant the crop the following spring. With the best cutting regime (double cut) and N rate (150 kg N ha^?1), 1514.4 and 2390.0 kg ha^?1 of dry leaves were produced, respectively, in 2014 and 2015. Leaf chlorophyll concentrations estimated by the SPAD (Soil and Plant Analysis Development)-502 chlorophyll meter and a NDVI (Normalized Difference Vegetation Index) carried out by the Field Scout CM 1000 spectroradiometer showed significant differences among N rates, proving to be good indicators of plant N nutritional status. Based on the leaf analysis, provisional sufficiency ranges for N are proposed, namely 25–35 g kg^?1 for mid-summer and 15–25 g kg^?1 for early autumn. The fluorescence of chlorophyll a and the transient fluorescence intensity performed by the OS-30p+ fluorometer failed to show any stress induced by no-N control treatments in comparison to N-treated plants.     

Effect of the simultaneous interaction among ascorbic acid, iron and pH on the oxidative stability of oil-in-water emulsions

The objective of this study was to demonstrate how different factors can simultaneously influence the oxidative stability of an oil-in-water emulsion, and how these factors can be used to enlarge the variation range of oxidation markers, expressed as peroxide value (PV) and TBARS. Initially, a Plackett-Burman design was used to screen seven factors (temperature, pH, and iron, copper, ascorbyl palmitate, ascorbic acid, and sodium chloride concentrations). A temperature elevation of 30 to 60 °C reduced PV and TBARS, a pH change from 3.0 to 7.0 increased PV and reduced TBARS, and the presence of ascorbic acid (1 mmol/L) had no significant effect on PV but increased TBARS (p < 0.05). Thus, the temperature was fixed at 30 °C, and an emulsion was formulated with different combinations of ascorbic acid, iron, and pH according to a central composite rotatable design. Regression models were fitted to PV and TBARs responses and optimized to get the higher values of both markers of oxidation. The optimized emulsion contained 1.70 mmol/L AH (ascorbic acid) and 0.885 mmol/L FeSO(4) · 7H(2)O (1.0 mmol/L Fe(2+)) at pH 5.51 and 30 °C. The range of variation observed for oxidation markers in the optimized emulsion model (PV, 0-4.27 mequiv/L; TBARS, 0-13.55 mmol/L) was larger than the variation observed in the nonoptimized model (PV, 0-1.05 mequiv/L; TBARS, 0-1.00 mmol/L). The antioxidant activity of six compounds (Trolox, α-tocopherol, caffeic acid, gallic acid, catechin, and TBHQ) was evaluated using the optimized emulsion conditions. After application of the Tukey HSD post hoc statistical test, the samples that were not different (p < 0.05) in the nonoptimized emulsions showed a significant difference in the optimized emulsions. Considering the importance of the interactions on oxidation studies, our model represents a significant improvement in a direct methodology that can be applied to evaluate natural compounds under different combination of factors.     

Analysis of Acid Alizarin Violet N Dye Removal Using Sugarcane Bagasse as Adsorbent

With the development of the textile industry, there has been a demand for dye removal from contaminated effluents. In recent years, attention has been directed toward various natural solid materials that are capable of removing pollutants from contaminated water at low cost. One such material is sugarcane bagasse. The aim of the present study was to evaluate adsorption of the dye Acid Violet Alizarin N with different concentrations of sugarcane bagasse and granulometry in agitated systems at different pH. The most promising data (achieved with pH 2.5) was analyzed with both Freundlich and Langmuir isotherms equations. The model that better fits dye adsorption interaction into sugarcane bagasse is Freundlich equation, and thus the multilayer model. Moreover, a smaller bagasse granulometry led to greater dye adsorption. The best treatment was achieved with a granulometry value lower than 0.21 mm at pH 2.50, in which the total removal was estimated at a concentration of 16.25 mg mL⁻¹. Hence, sugarcane bagasse proves to be very attractive for dye removal from textile effluents.