Polyethylene glycol and polyvinylpirrolidone effect on bacterial rRNA extraction and hybridization from cells exposed to tannins

In order to detect fluctuations in ruminal microbial populations due to forage tannins using 16S ribosomal RNA (rRNA) probes, recovery of intact rRNA is required. The objective of this work was to evaluate the effect of polyethylene glycol (PEG) and polyvinylpirrolidone (PVP) on extraction of bacterial rRNA, in the presence of tannins from tropical legume forages and other sources, that hybridize with oligonucleotide probes. Ruminococcus albus 8 cells were exposed to 8 g/L tannic acid or 1 g/L condensed tannins extracted from Acacia angustissima, banana (Musa sp.) skin, Desmodium ovalifolium, red grape (Vitis vinifera) skin and Inga edulis, or no tannins. Cells were rinsed with Tris buffer pH 7 containing either 8% PEG or 6% PVP prior to cell lysis. Total RNA samples rinsed with either PEG or PVP migrated through denaturing agarose gels. The 16S rRNA bands successfully hybridized with a R. albus species-specific oligonucleotide probe, regardless of tannin source. The effect of rinsing buffers on the density of 16S rRNA bands, as well as on the hybridization signals was compared. There were significant effects (P<0.01) when the controls were compared to either buffer treatments due to tannin type, buffer used and the interaction of tannin type and buffer. The significant interaction indicates the influence of tannin type on the parameters evaluated.

Toxicity of neem oil to Bemisia tabaci biotype B nymphs reared on dry bean

The objective of this work was to determine the most susceptible nymphal stage of Bemisia tabaci biotype B to neem (Azadirachta indica A. Juss.) oil applied to dry bean (Phaseolus vulgaris L.) in a screenhouse. A solution of commercial oil (Dalneem) extracted from neem seeds was sprayed directly on each nymphal instar at 0, 0.1, 0.25, 0.5, 1 and 2% concentrations for lethal concentration (LC) determination, and at 0, 0.5 and 1% concentrations for lethal time (LT) determination. The number of living and dead nymphs was recorded five days after spraying for LC determination, and daily during six days for LT determination. The LC50 estimated for fourth instar nymphs occurred at 0.56% concentration. For all instars, LC50 and LC95 were estimated at 0.32 and 2.78% concentrations, respectively. The estimated values of LT50 at 1% concentration were 2.46, 4.45, 3.02 and 6.98 days for the first to fourth instars, respectively. The LT50 occurred at five days for 0.5% and at four days for 1% concentration in all instars. A mortality rate of over 80% was observed on the 6th day for the first to third instars at 1% concentration. The first three nymphal stages were more susceptible to neem oil when compared to the fourth nymphal stage.

Predicting rapeseed oil content with near-infrared spectroscopy

The objective of this work was to establish a calibration equation and to estimate the efficiency of near-infrared reflectance (NIR) spectroscopy for evaluating rapeseed oil content in Southern Brazil. Spectral data from 124 half-sib families were correlated with oil contents determined by the chemical method. The accuracy of the equation was verified by coefficient of determination (R²) of 0.92, error of calibration (SEC) of 0.78, and error of performance (SEP) of 1.22. The oil content of ten genotypes, which were not included in the calibration with NIR, was similar to the one obtained by the standard chemical method. NIR spectroscopy is adequate to differentiate oil content of rapeseed genotypes.

Influence of rootstocks and pruning times on yield and on nutrient content and extraction in 'Niagara Rosada' grapevine

The objective of this work was to evaluate the influence of rootstocks and pruning times on yield and on nutrient content and extraction by pruned branches and harvested bunches of 'Niagara Rosada' grapevine in subtropical climate. The rootstocks 'IAC 766', 'IAC 572', 'IAC 313', 'IAC 571-6', and '106-8 Mgt' were evaluated. Treatments consisted of a combination between five rootstocks and three pruning times. At pruning, fresh and dry matter mass of branches were evaluated to estimate biomass accumulation. At harvest, yield was estimated by weighing of bunches per plant. Branches and bunches were sampled at pruning and at harvest, respectively, for nutrient content analysis. Nutrient content and dry matter mass of branches and bunches were used to estimate total nutrient extraction. 'Niagara Rosada' grapevine grafted onto the 'IAC 572' rootstock had the highest yield and dry matter mass of bunches, which were significantly different from the ones observed in 'Niagara Rosada'/'IAC 313'. 'Niagara Rosada' grafted onto the 'IAC 572' rootstock extracted the largest quantity of K, P, Mg, S, Cu, and Fe, differing from 'IAC 313' and 'IAC 766' in K and P extraction, and from '106-8 Mgt' in Mg and S extraction. Winter pruning results in higher yield, dry matter accumulation by branches, and total nutrient content and extraction.

Biometric analysis of protein and oil contents of soybean genotypes in different environments

The objective of this work was to identify by biometric analyses the most stable soybean parents, with higher oil or protein contents, cultivated at different seasons and locations of the state of Minas Gerais, Brazil. Forty-nine genotypes were evaluated in the municipalities of Viçosa, Visconde do Rio Branco, and São Gotardo, in the state of Minas Gerais, from 2009 to 2011. Protein and oil contents were analyzed by infrared spectrometry using a FT-NIR analyzer. The effects of genotype, environment, and genotype x environment interaction were significant. The BARC-8 soybean genotype is the best parent to increase protein contents in the progenies, followed by BR 8014887 and CS 3032PTA276-3-4. Selection for high oil content is more efficient when the crossings involve the Suprema, CD 01RR8384, and A7002 genotypes, which show high mean phenotypic values, wide adaptability, and greater stability to environmental variation.

Seed biometric parameters in oil palm accessions from a Brazilian germplasm bank

The objective of this work was to evaluate the morphological diversity of oil palm seeds and to cluster the accessions according to their morphological characteristics. Forty-one accessions from the oil palm germplasm bank of Embrapa Amazônia Ocidental were evaluated - 18 of Elaeis oleifera and 23 of E. guineensis. The groups were formed based on morphological characteristics, by principal component analysis. In E. oleifera, four groups were formed, tied to their region of origin, but with significant morphological differences between accessions from the same population. For tenera-type E. guineensis seeds, three widely divergent groups were formed, especially as to external parameters, which differentiated them from the other ones. The parameter endocarp thickness stood out in intra- and inter-population differentiation. For dura-type E. guineensis, three groups were formed, with larger seeds and thicker endocarps, which differed from all the other ones. The variability observed for seed characteristics in the analyzed accessions allows the establishment of different groups, to define strategies for genetic improvement.

Fish oil replacement by soybean oil in the diet of fat snook juveniles

The objective of this work was to evaluate fish oil replacement by soybean oil in diets, as for the effects on the performance and body composition of juveniles of fat snook (Centropomus parallelus). The experiment was carried out in a randomized block design, with three treatments (lipid sources) and six replicates, in a 60-day period. Fat snook juveniles (24.17±0.28g) were distributed in 18 experimental tanks of 200 L each, equipped with aeration and heating systems, under continuous water renovation (800% per day). Three isoproteic (44% CP) and isoenergetic (4,635 kcal CE kg-1) diets were formulated to comprise three replacement rates (0, 50, and 100%) of fish oil by soybean oil. Biometric analyses were done to evaluate fish performance, and two entire specimens from each replicate were used for body composition analyses. The zootechnical indices of weight gain (38.68±5.41 g), feed conversion (1.38±0.10), and specific growth at 1.70±0.18% weight gain per day were considered satisfactory. Lipid source substitution does not affect the performance and body composition of fat snook juveniles, which suggests that soybean oil can replace fish oil in diet formulation.

In vitro maintenance, under slow-growth conditions, of oil palm germplasm obtained by embryo rescue

The objective of this work was to evaluate the in vitro maintenance of oil palm (Elaeis guineensis and E. oleifera) accessions under slow-growth conditions. Plants produced by embryo rescue were subject to 1/2MS culture medium supplemented with the carbohydrates sucrose, mannitol, and sorbitol at 1, 2, and 3% under 20 and 25±2ºC. After 12 months, the temperature of 20°C reduced plant growth. Sucrose is the most appropriate carbohydrate for maintaining the quality of the plants, whereas mannitol and sorbitol result in a reduced plant survival.

Grouping sunflower genotypes for yield, oil content, and reaction to Alternaria leaf spot using GGE biplot

Abstract:The objective of this work was to evaluate the suitability of the multivariate method of principal component analysis (PCA) using the GGE biplot software for grouping sunflower genotypes for their reaction to Alternaria leaf spot disease (Alternariaster helianthi), and for their yield and oil content. Sixty-nine genotypes were evaluated for disease severity in the field, at the R3 growth stage, in seven growing seasons, in Londrina, in the state of Paraná, Brazil, using a diagrammatic scale developed for this disease. Yield and oil content were also evaluated. Data were standardized using the software Statistica, and GGE biplot was used for PCA and graphical display of data. The first two principal components explained 77.9% of the total variation. According to the polygonal biplot using the first two principal components and three response variables, the genotypes were divided into seven sectors. Genotypes located on sectors 1 and 2 showed high yield and high oil content, respectively, and those located on sector 7 showed tolerance to the disease and high yield, despite the high disease severity. The principal component analysis using GGE biplot is an efficient method for grouping sunflower genotypes based on the studied variables.

COMPARISON OF RNA EXTRACTION METHODS FOR Passiflora edulis SIMS LEAVES

ABSTRACT Functional genomic analyses require intact RNA; however, Passiflora edulis leaves are rich in secondary metabolites that interfere with RNA extraction primarily by promoting oxidative processes and by precipitating with nucleic acids. This study aimed to analyse three RNA extraction methods, Concert™ Plant RNA Reagent (Invitrogen, Carlsbad, CA, USA), TRIzol® Reagent (Invitrogen) and TRIzol® Reagent (Invitrogen)/ice -commercial products specifically designed to extract RNA, and to determine which method is the most effective for extracting RNA from the leaves of passion fruit plants. In contrast to the RNA extracted using the other 2 methods, the RNA extracted using TRIzol® Reagent (Invitrogen) did not have acceptable A260/A280 and A260/A230 ratios and did not have ideal concentrations. Agarose gel electrophoresis showed a strong DNA band for all of the Concert™ method extractions but not for the TRIzol® and TRIzol®/ice methods. The TRIzol® method resulted in smears during electrophoresis. Due to its low levels of DNA contamination, ideal A260/A280 and A260/A230 ratios and superior sample integrity, RNA from the TRIzol®/ice method was used for reverse transcription-polymerase chain reaction (RT-PCR), and the resulting amplicons were highly similar. We conclude that TRIzol®/ice is the preferred method for RNA extraction for P. edulis leaves.

Aplicação de SPME (Solid Phase Micro-Extraction) na análise de águas potáveis de três localidades do estado de São Paulo

The technique of solid phase microextraction (SPME) was used for the extraction of halogenated contaminants of water samples from three cities of the State of São Paulo and the extracts were submitted to gas chromatographic analysis with electron capture detection (GC-ECD). In the samples of water collected at the city of São Paulo the detected level of trihalomethanes (THM) expressed as the sum of chloroform, dibromochloromethane and dichlorobromomethane, were higher than the permissible limit established by the Brazilian regulation. In the samples collected at the two other cities the level of any of the three THM remained below the sensitivity of the ECD.

Extrações de óleos de sementes de citros e suas atividades sobre a formiga cortadeira Atta sexdens e seu fungo simbionte

Seeds Citrus oils (C. sinensis, C. limon and C. reticulata) extraction with hexane in a soxhlet apparatus and through supercritical fluid (CO2) were done. Besides triglycerides, the oils obtained with hexane comprised volatile compounds such as terpenes and fatty alcohols, esters, and aldehydes. However, the oils obtained by extraction with supercritical fluid presented only triglycerides. These results indicate that the extraction using supercritical fluid presents better selectivity. The activity of the oils on the development of the ant symbiotic fungus, Leucoagaricus gongylophorus, showed week activity and the topic insecticide assay showed better activity for the tangerine seed oil.

Avaliação do óleo essencial de Aloysia sellowii (Briquet) moldenke (Verbenaceae) do Sul do Brasil

This work was performed with the aim of evaluating the chemical variability among samples of Aloysia sellowii (Verbenaceae) collected in different geographical regions as well as the application of supercritical CO2 for obtaining essential oil. Thus, samples were collected in different localities and oils were isolated by hydrodistillation and supercritical CO2. Results showed existence of two chimiotypes in the species (cineole and sabinene). The supercritical CO2 extraction process was appropriate for extraction of A. sellowii essential oil at 40 ºC, 110 bar, 2 mL/min of flow and 10 min of extraction time.

Solid phase extraction of iron and lead in environmental matrices on amberlite xad-1180/pv

A solid phase extraction procedure using Amberlite XAD-1180/Pyrocatechol violet (PV) chelating resin for the determination of iron and lead ions in various environmental samples was established. The procedure is based on the sorption of lead(II) and iron(III) ions onto the resin at pH 9, followed by elution with 1 mol/L HNO3 and determination by flame atomic absorption spectrometry. The influence of alkaline, earth alkaline and some transition metals, as interferents, are discussed. The recoveries for the spiked analytes were greater than 95%. The detection limits for lead and iron by FAAS were 0.37 µg/L and 0.20 µg/L, respectively. Validation of the method described here was performed by using three certified reference materials (SRM 1515 Apple Leaves, SRM 2711 Montana Soil and NRCC-SLRS-4 Riverine Water). The procedure was successfully applied to natural waters and human hair.

Sistema aquoso bifásico: uma alternativa eficiente para extração de íons

Solvent extraction has been successfully applied to metal ion preconcentration and often meant the use of toxic organic diluents. However, regulatory pressure is increasingly focusing on the use and disposal of organic solvents, and thus the development of nonhazardous alternatives is important. In this review, we examine the application of aqueous biphasic systems (ABS) to extraction of ions, analyzing their potential and limitations and suggest that ABSs could be an efficient substitute for oil/water biphasic systems. ABSs are formed by mixing certain inorganic salts and water-soluble polymers, or by mixing two water-soluble polymers.