Evaluation of the Antioxidant, Antimicrobial, and Anti-Biofilm Effects of the Stem Bark, Leaf, and Seed Extracts from Hymenaea courbaril and Characterization by UPLC-ESI-QTOF-MS/MS Analysis.

Currently, biofilm-forming bacteria are difficult to treat by conventional antibiotic therapy and are, thus, becoming a clinical and epidemiological problem worldwide. Medicinal plants have been identified as novel alternative treatments due to their therapeutic and antimicrobial effects. In this context, the present study aimed to determine the total phenolic content, antioxidant capacity, and antimicrobial and anti-biofilm potential of nine extracts of Hymenaea courbaril (Fabaceae), popularly known as Jatobá. Furthermore, extracts that exhibited biofilm inhibitory activity against S. aureus (ATCC 25923) were selected for UPLC-HRMS/MS chemical analysis. Our results showed a high total phenolic content, mainly in the stem bark extract, and that the plant is rich in compounds with antioxidant activity. In the anti-biofilm analysis, leaf extracts stood out in comparison with chloramphenicol, with inhibition percentages of 78.29% and 78.85%, respectively. Through chemical analysis by UPLC-HRMS/MS, chrysoeriol-7-O-neohesperidoside, isorhamnetin-3-O-glucoside, and 3,7-di-O-methylquercetin were annotated for the first time in the leaves of H. courbaril. Therefore, these results showed the potential use of H. courbaril as an antioxidant and point to its use in antimicrobial therapy with an anti-biofilm effect.

Toxicological profile of the Hymenaea courbaril stem bark hydroalcoholic extract using in vitro bioassays and an alternative in vivo Caenorhabditis elegans model.

Hymenaea genus has been used in folk medicine in Brazil, but few studies investigated its toxicity profile. Thus, the aim of this study was to determine toxicological parameters of Hymenaea courbaril stem bark hydroalcoholic extract by utilizing three cell lines including murine macrophages (RAW 264.7), mouse fibroblast cells (L929) and human lung fibroblast (MRC-5), as well as Salmonella/microsome assay, and in vivo Caenorhabditis elegans model. The predominant detected phytoconstituents in the extract were coumarins, flavonoids, phenolics, tannins and saponins and by HPLC analysis, astilbin (AST) was found to be the main component. The DPPH assay demonstrated that H. courbaril hydroalcoholic extract exhibited potent antioxidant activity, with an IC50 of 3.12 µg/ml. The extract at concentrations of 400 and 800 µg/ml decreased cell viability 48 hr after treatment in L929 and MRC-5 cell lines. In the Raw 264.7 strain, just the highest concentration (800 µg/ml) lowered cell viability within 48 hr following exposure. The concentration of 100 µg/ml did not markedly affect cell viability in the trypan blue assay. In the alkaline comet assay the extract was found to be non-genotoxic. In the Ames test, the extract exhibited low mutagenic potential without metabolic activation, since only the highest concentrations produced an effect. H. courbaril extract only affected the survival of C. elegans at concentrations of 800 and 1600 µl/ml. These findings demonstrate that H. courbaril extract appears to exert low toxicity as evidenced in vitro and mutagenicity assays; however, the biological relevance of the response of C. elegans survival to safety assessments needs further studies.

Cerrado and Pantanal fruit flours affect gut microbiota composition in healthy and post-COVID-19 individuals: an in vitro pilot fermentation study.

Cerrado and Pantanal plants can provide fruits with high nutritional value and antioxidants. This study aims to evaluate four fruit flours (from jatobá pulp, cumbaru almond, bocaiuva pulp and bocaiuva almond) and their effects on the gut microbiota in healthy (HD) and post-COVID-19 individuals (PC). An in vitro batch system was carried out, the microbiota was analysed by 16S rRNA amplicon sequencing and the short-chain fatty acids ratio was determined. Furthermore, the effect of jatobá pulp flour oil (JAO) on cell viability, oxidative stress and DNA damage was investigated in a myelo-monocytic cell line. Beyond confirming a microbiota imbalance in PC, we identified flour-specific effects: (i) reduction of Veillonellaceae with jatobá extract in PC samples; (ii) decrease in Akkermansia with jatoba and cumbaru flours; (iii) decreasing trend of Faecalibacterium and Ruminococcus with all flours tested, with the exception of the bocaiuva almond in HD samples for Ruminococcus and (iv) increase in Lactobacillus and Bifidobacterium in PC samples with bocaiuva almond flour. JAO displayed antioxidant properties protecting cells from daunorubicin-induced cytotoxicity, oxidative stress and DNA damage. The promising microbiota-modulating abilities of some flours and the chemopreventive effects of JAO deserve to be further explored in human intervention studies.

In vivo evaluation of the toxic activity and genotoxicity of the Hymenaea courbaril L.'s resin in Drosophila melanogaster.

Due to the negative consequences carried by the usage of synthetic insecticides, a global interest into finding substitutes for these chemical compounds through natural products has arisen. When yielded to external attacks, plants generally produce metabolites to defend themselves. The physicochemical characteristics of this kind of compounds have allowed their usage as potential bioinsecticides. The Hymenaea courbaril L. (algarrobo) has proven to be a plant rich in metabolites with outstanding biological activity, in such a way that some of its extracts have been tested as insecticides. The goal of this study was to know the phytochemical composition of Hymenaea courbaril L.'s resin and perform evaluations in vivo of its toxic and genotoxic effects in the biological model Drosophila melanogaster. For this, two resin extracts were prepared and both a phytochemical analysis were carried out on them, having found in the ethanolic total extract the presence of terpenes, flavonoids and coumarins, while in the partial ethanolic extract only presence of terpenes and flavonoids was found. Drosophila larvae were submitted to different concentrations of the extracts and both the survival and the sexual ratio were evaluated, finding that larvae are more sensitive to the partial ethanolic extract. Subsequently, the induction of somatic mutation and mitotic recombination (SMART) was evaluated in the flies' eyes. The most significant affectations at a genotoxic level were found when larvae were tested with the partial extract, indicating that possibly the coumarins absence makes this insect more susceptible to damages at a genetic material level.

Caryophyllene Oxide, the Active Compound Isolated from Leaves of Hymenaea courbaril L. (Fabaceae) with Antiproliferative and Apoptotic Effects on PC-3 Androgen-Independent Prostate Cancer Cell Line.

Cancer treatment frequently carries side effects, therefore, the search for new selective and effective molecules is indispensable. Hymenaea courbaril L. has been used in traditional medicine in South America to treat several diseases, including prostate cancer. Leaves' extracts from different polarities were evaluated using the 3-(4,5-methyl-thiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) cell viability assay to determine the cytotoxicity in prostate p53-null cells, followed by bio-guided fractionations to obtain the most cytotoxic fraction considering the selectivity index. The most cytotoxic fraction was analyzed by GC/MS to identify the active compounds. The majority compound, caryophyllene oxide, induced early and late apoptosis, depolarized the mitochondrial membrane, leading to several morphological changes and shifts in apoptotic proteins, and caspases were evidenced. Depolarization of the mitochondrial membrane releases the pro-apoptotic protein Bax from Bcl-xL. The apoptosis process is caspase-7 activation-dependent. Caryophyllene oxide is a safe anti-proliferative agent against PC-3 cells, inducing apoptosis with low toxicity towards normal cells.

Prospection of Fungal Lignocellulolytic Enzymes Produced from Jatoba (Hymenaea courbaril) and Tamarind (Tamarindus indica) Seeds: Scaling for Bioreactor and Saccharification Profile of Sugarcane Bagasse.

The lignocellulosic biomass comprises three main components: cellulose, hemicellulose, and lignin. Degradation and conversion of these three components are attractive to biotechnology. This study aimed to prospect fungal lignocellulolytic enzymes with potential industrial applications, produced through a temporal analysis using Hymenaea courbaril and Tamarindus indica seeds as carbon sources. α-L-arabinofuranosidase, acetyl xylan esterase, endo-1,5-α-L-arabinanase, ß-D-galactosidase, ß-D-glucosidase, ß-glucanase, ß-D-xylosidase, cellobiohydrolase, endoglucanase, lichenase, mannanase, polygalacturonase, endo-1,4-ß-xylanase, and xyloglucanase activities were determined. The enzymes were produced for eight filamentous fungi: Aspergillus fumigatus, Trametes hirsuta, Lasiodiplodia sp., two strains of Trichoderma longibrachiatum, Neocosmospora perseae, Fusarium sp. and Thermothelomyces thermophilus. The best producers concerning enzymatic activity were T. thermophilus and T. longibrachiatum. The optimal conditions for enzyme production were the media supplemented with tamarind seeds, under agitation, for 72 h. This analysis was essential to demonstrate that cultivation conditions, static and under agitation, exert strong influences on the production of several enzymes produced by different fungi. The kind of sugarcane, pretreatment used, microorganisms, and carbon sources proved limiting sugar profile factors.

Concentración foliar de macro- y micro-nutrimentos en cuatro leguminosas maderables del trópico estacionalmente seco de Costa Rica / Foliar concentration of macro- and micronutrients in four woody legumes from a tropical dry-seasonal forest in Costa Rica

Resumen La productividad de las plantaciones forestales con especies nativas se puede incrementar si se conocen bien los requerimientos nutricionales de cada una de las especies. Estos requerimientos pueden variar de acuerdo a las estrategias ecológicas de las especies y a las adaptaciones que tienen, como por ejemplo la capacidad de fijar nitrógeno. En el presente trabajo se presentan los "rangos satisfactorios" de concentración foliar de macronutrimentos y micronutrimentos para cuatro leguminosas del trópico estacionalmente seco de Guanacaste, Costa Rica. Se analizó la concentración foliar de N, P, Ca, Mg, K, S, Fe, Mn, Cu, Zn y B, Na en las especies Dalbergia retusa (cocobolo), Enterolobium cyclocarpum (guanacaste), Hymenaea courbaril (guapinol) y Schizolobium parahyba (gallinazo). La concentración de nutrimentos varió con las especies y según la capacidad de fijar nitrógeno o no y dentro de cada una de esas categorías de acuerdo a la tasa de crecimiento. La concentración total de nutrimentos fue mayor para las especies fijadoras de N que para las no fijadoras y dentro de cada uno de esos grupos la concentración total de nutrimentos fue mayor para las especies de crecimiento rápido comparado con aquellas de crecimiento más lento. De acuerdo a la cantidad de nutrimentos observados en las hojas, se recomienda que H. courbaril y D. retusa deberían ser sembradas en suelos relativamente pobres de nutrimentos, mientras que E. cyclocarpum y S. parahyba en suelos más fértiles. Para D. retusa e H. courbaril se presenta información nueva sobre concentraciones y se mejora la misma para algunos elementos no mencionados en la literatura para E. cyclocarpum y S. parahyba.

Abstract The information about nutritional aspects of trees in the tropical dry forest is scarce. This investigation aims to establish a normal range of foliar nutrient concentration to four forestry species as related to their growth rates and biological nitrogen fixation capacity in the seasonally dry ecosystem at Guanacaste, Costa Rica. Foliar samples for four species: Dalbergia retusa (cocobolo), Enterolobium cyclocarpum (guanacaste), Hymenaea courbaril (guapinol or jatobá) and Schizolobium parahyba (gallinazo or pachaco), all belonging to the Fabaceae family, were analyzed for N, P, Ca, Mg, K, S, Fe, Mn, Cu, Zn, B and Na. Nutrient concentration varied with species according to their ability to fix nitrogen and their rate of growth. The overall concentration of macronutrients was higher in nitrogen-fixing species compared with non-fixing species, and within each one of those groups, the species with higher growth rate presented more macronutrients than species with low growth rate. According to the concentration of foliar nutrients accumulated in the foliage, we recommend that H. courbaril and D. retusacould be grown in the region soils with medium fertility and E. cyclocarpum and S. parahyba in the more fertile soils. Values ranges of foliar concentrations considered as satisfactory presented in this work are new for D. retusa and H. courbaril and improve the scarce information available for E. cyclocarpum and S. parahyba. Rev. Biol. Trop. 66(3): 969-983. Epub 2018 September 01.

Phytoremediation of Brazilian tree species in soils contaminated by herbicides.

The phytoremediation potential of Brazilian trees species Calophyllum brasiliense, Eremanthus crotonoides, Hymenaea courbaril, Inga striata, and Protium heptaphyllum was investigated for contaminated soils containing ametryn and hexazinone. Visual injury, chlorophyll content, plant height, leaf temperature, leaf number, and water-efficient use were evaluated. Residual herbicides at soil substrates were analyzed by LC/MS. Among the species C. brasiliense and H. courbaril were tolerant to both herbicides. P. Heptaphyllum presented tolerance at ametryn treatment. E. crotonoides, I. striata, and P. heptaphyllum died in hexazinone treatment. A high content of residual ametryn was found for E. crotonoides. In ametryn treatment, residual herbicide has decreased for C. brasiliense and E. crotonoides species. C. brasiliense highlighted among others becoming a good agent for phytoremediation of soils contaminated with traces of ametryn and hexazinone.

Preparation of biosorbents from the Jatoba (Hymenaea courbaril) fruit shell for removal of Pb(II) and Cd(II) from aqueous solution.

In this study, the biosorption properties of Jatoba (Hymenaea courbaril) fruit shell for removal of Pb(II) and Cd(II) ions from aqueous solutions, and its potential as a low-cost biosorbent for water treatment, were investigated. The Jatoba fruit shell (JBin) was subjected to different treatments with heated water (JBH2O) and sodium hydroxide (JBNaOH) to modify its surfaces and improve its adsorption properties. The chemical modification of the surfaces of the resulting materials was confirmed by analyzing the compositions and structural features of the raw material and the chemically treated materials using SEM, FTIR, 13C NMR, and pHpzc. The ability of the biosorbents to remove the metal ions was investigated with batch adsorption procedures. The adsorption data were then examined in detail by applying adsorption models of Langmuir, Freundlich, and Dubinin-Radushkevich. The results showed that the experimental data were best described by the Langmuir model for the Pb-JBin and Cd-JBNaOH systems, the Freundlich model for the Pb-JBH2O and Pb-JBNaOH systems, and the Dubinin-Radushkevich model for Cd-JBin and Cd-JBH2O systems. The maximum adsorption capacities of JBNaOH obtained using the Langmuir model reached values of 30.27 and 48.75 mg g-1 for Cd(II) and Pb(II), respectively. The adsorption kinetic studies showed that the pseudo-second-order model was the best fitted to the experimental data, and adsorptions for Pb-JBH2O and Cd-JBH2O are controlled by intraparticle diffusion mechanism.

First isolation of Cryptococcus neoformans genotype VNI MAT-alpha from wood inside hollow trunks of Hymenaea courbaril.

Cryptococcal infection is transmitted by the inhalation of Cryptococcus spp. propagules. Information about the Cryptococcus species inhabiting plants might be clinically relevant due to the epidemiological role of these habitats as possible sources of human infection. The aim of this study was to increase the knowledge about the environmental occurrence of cryptococcosis agents. Hollow tree vegetal debris of nine plant species was sampled quarterly over a 12-month period. Melanized colonies were screened for Cryptococcus neoformans and C. gattii by biochemical tests, followed by URA5-RFLP molecular analysis, M13 fingerprinting assays, and mating-typing with the specific a and α primers. The susceptibility to fluconazole of all of the confirmed species colonies was determined using the AFST-EUCAST broth dilution method. We found that the typical Brazilian flora tree Hymenaea courbaril yielded a high cryptococcal burden (median, 10(2) CFU/g) during the summer, autumn and winter seasons. C. neoformans VNI molecular type MAT alpha was identified in all of the samples. The fingerprinting analyses showed great molecular variability with no correlation with the susceptibility profile to fluconazole (MIC range 4 to ≥64 mg/l). To our knowledge, this study is the first describing the association between C. neoformans and Hymenaea courbaril. These observations extend the known geographic distribution of and substantiate a new urban environmental niche for C. neoformans and also emphasize the genetic diversity of the environmental C. neoformans VNI molecular type isolates.

Structure and rheological properties of a xyloglucan extracted from Hymenaea courbaril var. courbaril seeds.

Hymenaea courbaril var courbaril seed xyloglucan was efficiently extracted with 0.1M NaCl, followed by ethanol precipitation (yield=72±5% w/w). Its amorphous structure was identified by the pattern of X-ray diffraction. The monosaccharide composition was determined by GC/MS analysis of the alditol acetates and showed the occurrence of glucose:xylose:galactose:arabinose (40:34:20:6). One-(1D) and two-dimensional-(2D) NMR spectra confirmed a central backbone composed by 4-linked ß-glucose units partially branched at position 6 with non-reducing terminal units of α-xylose or ß-galactose-(1→2)-α-xylose disaccharides. The xyloglucan solution was evaluated by dynamic light scattering and presents a polydisperse and practically neutral profile, and at 0.5 and 1.0% (w/v) the solutions behave as a viscoelastic fluid. The polysaccharide did not show significant antibacterial or hemolytic activities. Overall our results indicate that xyloglucan from H. courbaril is a promising polysaccharide for food and pharmaceutical industries.

Analysis of xyloglucans by ambient chloride attachment ionization tandem mass spectrometry.

Xyloglucan oligomers obtained upon enzyme digestion from Hymenaea courbaril, Arabidopsis Columbia-0 and mur3 were ionized and analyzed by using chloride anion attachment electrospray ionization (ESI) and tandem mass spectrometry. MW determination and structural elucidation of several xyloglucan oligomers was performed directly from the mixture solutions without sample pretreatment or derivatization. Sodium cation attachment was used to determine the number of xyloglucans present in the mixtures and their MWs. However, tandem mass spectrometry results showed that structure elucidation based on the sodium adducts is ambiguous. Chloride anion also forms stable adducts with these xyloglucans upon ESI. These adducts can be readily identified due to the chlorine isotope pattern. The mass spectral profile of xyloglucans obtained for the mixtures matches the HPAEC results, thus validating this methodology for the determination of the xyloglucan composition and the MW of each xyloglucan. Upon collisional activation in MS(2) experiments, the chloride anion adducts readily lose HCl, which helps verify the molecular weight of each xyloglucan. Isolating the resulting anion (deprotonated oligomer) and subjecting it to further collision-activated dissociation experiments (MS(n); n=3-4) yields useful structural information that allows the differentiation between isomeric anions and hence determination of the sequence of the xyloglucan oligomers. The deprotonated oligomers fragment by a stepwise loss of sugar units from the reducing end.

Phytochemical study guided by the myorelaxant activity of the crude extract, fractions and constituent from stem bark of Hymenaea courbaril L.

ETHNOPHARMACOLOGICAL RELEVANCE: Hymenaea courbaril L. (Caesalpinoideae) is used in Brazilian folk medicine to treat anemia, kidney problems, sore throat and other dysfunctions of the respiratory system, such as bronchitis and asthma, although such properties are yet to be scientifically validated. AIM OF THE STUDY: In order to give a scientific basis to support the traditional use of Hymenaea courbaril, this study was designed to evaluate antioxidant, myorelaxant and anti-inflammatory properties of the ethanol extract from stem bark and its fractions. The myorelaxant effect of astilbin, a flavonoid isolated from the bioactive ethyl acetate fraction (EAF), has also been evaluated. MATERIAL AND METHODS: In the present study ethanol extract from stem bark (EEHC) and fractions were analyzed using bioassay-guided fractionation. The following activities were investigated: antioxidant by 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay, myorelaxant on rat tracheal smooth muscle, and anti-inflammatory using ovalbumin-induced leukocytosis and airway hyperresponsiveness in rats. RESULTS: The results of the present investigation show that the whole extract of Hymenaea courbaril and some of its fractions strongly scavenged DPPH radical. The extract showed myorelaxant activity on rat trachea, being EAF its highest efficient fraction. Bio-guided study allowed the isolation of astilbin, a well-known flavonoid. The activity induced by this compound indicates that it may be partly responsible for the myorelaxant effect of EAF. EAF reduced contractions that depended on divalent cation inflow through voltage-operated Ca(2+) channels (VOCCs) or receptor-operated Ca(2+) channels (ROCCs), but it was more potent to inhibit VOCC- than ROCC-dependent contraction induced by Ca(2+) addition in ACh-enriched Ca(2+)-free medium. Oral pretreatment of antigen-challenged animals with EAF prevented airway hyperresponsiveness on KCl-induced contraction and reduced the number of total white cells, particularly eosinophils and neutrophils in bronchoalveolar lavage. CONCLUSIONS: This study provided scientific basis that Hymenaea courbaril presents potential antioxidant, myorelaxant and anti-inflammatory actions, which support its use in folk medicine to treat inflammatory airway diseases.

Crescimento de Hymenaea courbaril L. var. stilbocarpa (Hayne) Lee et Lang. e Enterolobium contortisiliquum (Vell) Morong (Leguminosae) sob diferentes níveis de sombreamento / Growth of Hymenaea courbaril L. var. stilbocarpa (Hayne) Lee et Lang. e Enterolobium contortisiliquum (Vell) Morong (Leguminosae) under different shading levels

Hymenaea courbaril L. var. stilbocarpa (Hayne) Lee et Lang. é uma espécie clímax tolerante a sombra, ao passo que Enterolobium contortisiliquum (Vell.) Morong. é uma espécie pioneira. O desenvolvimento destas espécies pode refletir a habilidade de adaptação aos diferentes fatores ambientais (luz, água e temperatura) no local em que estão crescendo. O suprimento inadequado de um desses fatores pode reduzir o vigor da planta e limitar seu desenvolvimento. O presente trabalho teve como objetivo avaliar os efeitos do nível de sombreamento no crescimento e a concentração de pigmentos fotossintéticos em duas espécies de leguminosas arbóreas, Hymenaea courbaril L. var. stilbocarpa (Hayne) Lee et Lang. e Enterolobium contortisiliquum (Vell.) Morong. O experimento foi conduzido no Setor de Olericultura do Centro Universitário Luterano de Ji-Paraná (CEULJI/ULBRA)/Rondônia. Durante a formação das mudas, ambas as espécies foram expostas a quatro tratamentos de sombra: 0 % (controle - sol pleno); 30 %; 50 % e 80 %. Cada tratamento foi constituído com três repetições de cada espécie; o delineamento experimental foi inteiramente casualisado. Quatro meses após a semeadura, as seguintes análises foram realizadas: número de folhas, altura da planta, comprimento do sistema radicular, massa seca total e concentração de pigmentos fotossintéticos. O tratamento sob sol pleno afetou negativamente o crescimento de ambas as espécies. As mudas crescidas sob 50% e 80% apresentaram melhor desenvolvimento. Conforme o aumento do sombreamento houve um decréscimo na razão clorofila a/b e um aumento nas concentrações de clorofila total e carotenóides totais.

Hymenaea courbaril L. var. stilbocarpa (Hayne) Lee et Lang. is a clímax shadow tolerant specie and Enterolobium contortisiliquum (Vell.) Morong., by the other hand, is considered as a pioneer specie. The development of these species may reflect its adaptation ability to different environmental elements (light, water and temperature) at its growth site. An inadequate supply of some of these elements may reduce plant vigor and limit its development. This work aimed to evaluate the effects of shade intensity on the growth and photosynthetic pigment concentration of two arboreal legumes that are typical to northern Brazil, namely, the Hymenaea courbaril L. var. stilbocarpa (Hayne) Lee et Lang. and the Enterolobium contortisiliquum (Vell.) Morong. The experiment was carried out at the Centro Universitário Luterano de Ji-Paraná (CEULJ/ULBRA)/Rondônia. During the formation of the seedlings, both species were exposed to four treatments of shade: 0 % (control - without shade); 30 %; 50 % e 80 %. Each treatment was carried out with three replications for each species; the experimental design was completely randomized. Four month after sowing, the following analyses were carried out: number of leaves, plant height, roots length, dry mass and photosynthetic pigment concentration. The treatment under 0% shade impaired the seedling growth of both species. The most favorable treatments were 50 % and 80 % shade. According to the shade intensity, the chlorophyll a/b ratio decreased and the chlorophyll and carotenoids increased.

Produção de biomassa e estrutura do pasto de capim-andropogon em sistema silvipastoril e monocultura / Biomass production and pasture structure of Andropogon grass in a silvopastoral system and in monoculture

A produção de biomassa e as características estruturais do capim-andropogon foram avaliadas nas idades de rebrota de 35, 49 e 63 dias, em três ambientes de um sistema silvipastoril: sob a copa de pau-d'arco, sob a copa de jatobá, em área aberta; local fora da copa de qualquer árvore e em sistema de monocultura. O delineamento experimental foi o de parcelas subdivididas com cinco repetições. As parcelas foram representadas pelos ambientes e as subparcelas, pelas idades de rebrota. A luminosidade sob pau-d'arco, sob jatobá e em área aberta foi de 74, 62 por cento e 82 por cento, respectivamente, das áreas a pleno sol. Houve interação ambiente versus idade de rebrota. O sombreamento sob a copa de pau-d'arco e de jatobá não afetou a produção de matéria seca do capim-andropogon, contudo reduziu a altura do pasto aos 63 dias de rebrota. O percentual de folhas da forragem aos 35 dias de rebrota foi mais elevado na monocultura. Nas outras idades de rebrota, os quatro ambientes não diferiram entre si.

The biomass production and pasture structure of Andropogon grass were evaluated at 35, 49, and 63 days of regrowth, in three silvopastoral environments: under Pau-d'Arco and Jatobá trees, in open field and in monoculture system. The experiment was designed in split plot with five replications, the plots were the environments and the split plots were the regrowth ages. The luminosities under Pau-d'Arco, Jatobá, and open field were: 74, 62, and 82 percent, respectively, relative to the areas under full sun. There was interaction among environments and ages of regrowth. The shading under the canopy of Pau-d'Arco and Jatobá did not affect the production of dry matter of Andropogon grass; however, reduced the height of the grass at 63 days of regrowth. The percentage of leaves of forage at 35 days of regrowth was higher in monoculture. At other ages of regrowth, the four environments did not differ among themselves.

An efficient and rapid DNA minipreparation procedure suitable for PCR/SSR and RAPD analyses in tropical forest tree species

An efficient and rapid DNA minipreparation modified method for frozen samples was developed for five tropical tree species: Copaifera langsdorffii, Hymenaea courbaril, Eugenia uniflora, Tabebuia roseo alba and Cariniana estrellensis. This procedure that dispenses the use of liquid nitrogen, phenol and the addition of proteinase K, is an adaptation of the CTAB-based DNA extraction method. The modifications included the use of PVP to eliminate the polyphenols, only one chloroform-isoamyl alcohol step and the addition of RNase immediately after extraction with chloroform. The yields of the DNA samples ranged from 25.7 to 42.1 µg from 100 mg leaf tissue. The DNA samples extracted by this method were successfully used for PCR (SSR and RAPD) analyses in these five and other twelve tropical tree species.

Este trabalho teve como objetivo otimizar um protocolo econômico, rápido e eficaz de minipreparação de DNA genômico, para as espécies florestais Copaifera langsdorffii (Óleo de Copaíba), Hymenaea courbaril (Jatobá), Eugenia uniflora (Pitanga), Tabebuia roseo alba (Ipê Branco) e Cariniana estrellensis (Jequitibá Branco). Este método é uma adaptação da técnica de extração CTAB de Doyle e Doyle (1990), o qual consiste principalmente na adição de PVP para eliminar polifenoles, somente uma etapa de extração com clorofórmio-álcool isoamílico e a adição da RNase A imediatamente após a extração com clorofórmio. O método também dispensa o uso de nitrogênio líquido, o uso do fenol e a adição de proteinase K. Os DNAs das espécies florestais extraídos apresentaram alto rendimento e boa qualidade, com rendimento de 25.7 a 42.1 µg de DNA a partir de 100 mg de tecido foliar congelado. Com este protocolo, em apenas 1 dia de trabalho, uma pessoa pode completar o isolamento do DNA de aproximadamente 50 amostras de folhas (dependendo da capacidade da centrífuga). O DNA obtido pode ser usado para métodos de análise baseados em PCR (SSR e RAPD).

Funcionalidad de una mezcla de gomas de acasia glomerosa, Enterolobium cyclocarpum e Hymenaea courbaril en la preparación de helados de bajo contenido calórico / Functionality of a mixture of gums from acacia glomerosa, Enterolobium cyclocarpum and Hymenea courbaril in the low fat ice-cream preparation

Acacia glomerosa, Anterolobium cyclocarpum e Hymenaea courbaril, especies arbóreas ampliamente disemindas en Venezuela producen gomas con buen rendimiento. La mezcla de estas gomas se ensayó, como estabilizante en la preparación de helados de bajo contenido calórico. La viscosidad, el porcentaje de derretido y las propiedades sensoriales fueron determinados. Estas características físico-químicas se compararon con las exhibidas por los productos obtenidos con base en mezcla de gomas comerciales. La mezcla de gomas de las especies estudiadas incrementó la viscocidad del sistema, por lo tanto, favoreció una excelente incorporación y una distribución uniforme del aire; aportó textura y estabilidad durante el almacenamiento; mejoró la capacidad de expansión de la espuma y las propiedades de derretido. Por otra parte, aportó buenas características sensoriales, como la indica el alto puntaje alcanzado; apariencia (7.78), cremosidad (7.62), sabor (8.01) y bajo puntaje en presencia de cristales de hielo (3.06). Este trabajo evidenció la funcionalidad como estabilizante de la mezcla de gomas investigadas en la preparación de helados bajo en calorías.

Acacia glomerosa, Enterolobium cyclocarpum and Hymenaea courbaril, species widely disseminated in Venezuela, South America, produce gum with good yield. The mixture of these gums was tested as stabilizer in the low fat ice cream preparation. Viscosity, overrun, foam expansion, meltdown, shape factor and sensory properties were determined. These quality characteristics were compared with those exhibited by the product obtained with a mixture of commercial gums. The mixture tested provided the suitable viscosity for the ice cream mix with the corresponding overrun and texture. It gave better foaming properties and air incorporation. On the other hand, it was demonstrated the good sensory attributes by the highest score i.e. appearance (7.78), creaminess (7.62), flavor (8.01) and lowest score of iciness (3.06). This work showed good functionality as stabilizer of the mixture of gums investigated in the low fat ice cream preparation.