Organic compounds from plant extracts and their effect on soil phosphorus availability

The objective of this work was to evaluate the effect of organic compounds from plant extracts of six species and phosphate fertilization on soil phosphorus availability. Pots of 30 cm height and 5 cm diameter were filled with Typic Hapludox. Each pot constituted a plot of a completely randomized design, in a 7x2 factorial arrangement, with four replicates. Aqueous extracts of black oat (Avena strigosa), radish (Raphanus sativus), corn (Zea mays), millet (Pennisetum glaucum), soybean (Glycine max), sorghum (Sorghum bicolor), and water, as control, were added in each plot, with or without soluble phosphate fertilization. After seven days of incubation, soil samples were taken from soil layers at various depths, and labile, moderately labile and nonlabile P fractions in the soil were analysed. Plant extracts led to an accumulation of inorganic phosphorus in labile and moderately labile fractions, mainly in the soil surface layer (0-5 cm). Radish, with a higher amount of malic acid and higher P content than other species, was the most efficient in increasing soil P availability.

Nymphicidal effect of vegetal extracts of Annona mucosa and Anonna crassiflora (Magnoliales, Annonaceae) against rice stalk stink bug, Tibraca limbativentris (Hemiptera, Pentatomidae)

This study aimed to verify the chloroform-methanol nymphicidal action of extracts of Annona mucosa leaves and seeds and of A. crassiflora seeds on second instar nymphs of rice stalk stink bug, Tibraca limbativentris. For each extract the concentrations of 0.5%, 1.0%, 2.0%, 4.0%, 8.0%, and two control treatments (water and Tween80®) were used. The results show that the seed extracts of A. mucosa and A. crassiflora have insecticidal activity against the T. limbativentris nymphs with statistical significance for all concentrations when compared with controls. The seed extract of A. mucosa showed the higher toxicity with greater than 75% mortality at a concentration of 1.0% in the first 24 h after application. The leaf extract of A. mucosa presented the lowest toxicity with no more than 40% mortality. The seed extract of A. crassiflora showed intermediate toxicity among all the tested extracts, and the nymph's mortality exceeded 80% for the highest concentration after 120 h of application. Considering these results, we were able to observe that the seeds extract of A. mucosa may be an alternative for the control of bed bug nymphs T. limbatriventris, especially for small producers.

Development and validation of a LC-method for the determination of phenols in a pharmaceutical formulation containing extracts from Stryphnodendron adstringens

A sensitive RP-HPLC method with UV detection successfully measured phenol(s) in an ointment containing 3% Stryphnodendron adstringens extract. Chromatography used acetonitrile (0.05% trifluoroacetic acid):water (0.05% trifluoroacetic acid) (v/v), flow rate 0.8 mL min-1. Quantitation was accomplished by the external-standard method. Linearity for 2.00 to 16.00 μg mL-1 (gallic acid) and 1.14 to 18.24 μg mL-1 (gallocatechin) was established. Intra- and inter-day precision levels were under 5%. LOD and LOQ were 0.231 and 0.770 μg mL-1 (gallic acid) and 0.151 and 0.504 μg mL-1 (gallocatechin), respectively. Determination of phenols was unaffected by product excipients.

Simultaneous determination of four phenolic compounds in extracts of aerial parts of Ipomoea pes-caprae (L.) R. Br. (Convolvulaceae) by HPLC-UV

Among other applications, Ipomoea pes-caprae is popularly used to treat jellyfish stings, supporting the development of a product for dermatological use. Hydroethanolic spray-dried extract was chosen for the further development of phytomedicines, and a stability-indicative HPLC-UV method was developed and validated for the determination of isoquercitrin and isochlorogenic acids A, B and C. The method was developed using a C18 column (250 x 4.6 mm, 5 µm) with an acetonitrile:water mobile phase at pH 3.0 in a gradient run. The four constituents and other unidentified components of the extract were appropriately resolved without interference of degradation products after stress tests (acid, alkali, neutral, oxidant, photolysis). The method showed linearity in the isoquercitrin concentration range from 5.0-50.0 µg mL-1, with adequate precision (RSD% < 2.5% for the intra- and inter-day studies), accuracy (recovery of 100.0 ± 2.0%), and robustness. Both the herbal drug and spray-dried extract of I. pes-caprae were subjected to stability studies in accelerated and long-term conditions over four months. The samples maintained their characteristics and marker contents (< 10% of variation).

Use of botanical extracts, cassava wastewater and nematicide for the control of root-knot nematode on carrot

Aqueous extracts of several plant species have shown promising in controlling root-knot nematode, Meloidogyne incognita (Kofoid & White), one of the most limiting agents for carrot cultivation. The current study evaluated the effect of aqueous extracts from seven botanical species applied to 40, 50, 60, 70 and 80 days after sowing 'Nantes' carrots in soil infested with root-knot nematode. Three other treatments included cassava wastewater, distilled water (control), which were applied in the same periods of the extracts application, in addition to carbofuran 50G (80Kg/ha), which was applied once at 60 days after carrot sowing. Evaluations were performed at 90 days after inoculation to determine shoot and root fresh weight, as well as the diameter and the length of principal roots and the number of galls on primary and secondary roots. Plants treated with cassava wastewater, extracts of Ricinus communis L. seeds, Crotalaria juncea L. seeds, R. communis leaves + branches + fruits, Chenopodium ambrosioides L. leaves + branches + inflorescences and Azadirachta indica A. Juss. seeds showed the highest rates of total weight (root + shoot) and shoot weight. The extract of R. communis leaves + branches + fruits provides the highest total root weight and principal root diameter. Cassava wastewater and extracts of R. communis seeds provided the highest principal root weight. The extract of R. communis seeds and cassava wastewater can be considered promising for the alternative control of M. incognita.

Allelopathic interference of Sapindus saponaria root and mature leaf aqueous extracts on diaspore germination and seedling growth of Lactuca sativa and Allium cepa

Sapindus saponaria (soapberry) is a species that presents a great diversity of chemical compounds, such as saponins; however, few studies have examined the allelopathic effect of this species. Therefore, this study provides an evaluation of the allelopathic potential of aqueous extracts of the roots and mature leaves of S. saponaria on the germination of diaspores and seedlings growth of lettuce (Lactuca sativa) and onion (Allium cepa). The aqueous extract was prepared in the proportion of 100 g of dry plant material in 1,000 mL of distilled water (a concentration of 10% w v-1), and diluted with distilled water to 7.5, 5.0 and 2.5% concentrations. The mature leaf extracts caused delay and decrease in the germination process of the lettuce and onion diaspores, with inhibitory effect concentration-dependent, while the root extracts showed no allelopathic effects on the germination process. Both extracts caused abnormalities and inhibited the growth of shoot and root seedlings.

Inotropic effects of extracts of Psidium guajava L. (guava) leaves on the guinea pig atrium

Many pharmacological effects have been ascribed to extracts of Psidium guajava L. (guava) leaves. However, in spite of its widespread use in Brazilian folk medicine and a reasonable number of scientific reports about it, we could not find any study dealing with its action on the mammalian myocardium. In the present study, by measuring isometric force, we observed that the crude extract of P. guajava (water-alcohol extract obtained by macerating dry leaves) depresses the guinea pig atrial contractility in a concentration-dependent fashion (N = 8 hearts, 15 trials). The compound with cardiac activity was concentrated by extraction in a Soxhlet apparatus using 17 M glacial acetic acid after removing the less polar fractions (hexane, chloroform, acetone, ethanol and methanol), suggesting that this compound is a highly polar substance. In the isolated guinea pig left atrium the acetic acid fraction (10-800 mg/l) of P. guajava 1) reversibly decreased myocardial force in a concentration-dependent fashion (EC50 = 0.07g/l, N = 5 hearts, 9 trials, P<0.05), 2) increased the atrial relaxation time measured at 20% of the force amplitude up to 35% (91 ± 15 to 123 ± 30 ms, N = 3 hearts, 6 trials, P<0.05), 3) abolished the positive staircase effect (Bowditch phenomenon) in a concentration-dependent fashion suggesting a decrease of the cellular inward calcium current (N = 4 hearts, 8 trials, P<0.05), and 4) its inotropic effect was abolished by cholinergic receptor blockade with 1.5 mM atropine sulfate, indicating a cholinergic involvement in the mechanism of action of the extract (N = 7 hearts, 15 trials, P<0.05). The acetic acid extract was 20 times more potent than crude extract (EC50 = 1.4 g/l). The results showed that extracts from P. guajava leaves depress myocardial inotropism.

Immunomodulatory properties of Alternanthera tenella Colla aqueous extracts in mice

Plants from the genus Alternanthera are thought to possess antimicrobial and antiviral properties. In Brazilian folk medicine, the aqueous extract of A. tenella Colla is used for its anti-inflammatory activity. The present study investigated the immunomodulatory property of A. tenella extract by evaluating the antibody production in male albino Swiss mice weighing 20-25 g (10 per group). The animals received standard laboratory diet and water ad libitum. The effect of A. tenella extract (5 and 50 mg/kg, ip) was evaluated in mice immunized with sheep red blood cells (SRBC 10%, ip) as T-dependent antigen, or in mice stimulated with mitogens (10 µg, Escherichia coli lipopolysaccharide, LPS, ip). The same doses (5 and 50 mg/kg, ip) of A. tenella extract were also tested for antitumor activity, using the Ehrlich ascites carcinoma as model. The results showed that 50 mg/kg A. tenella extract ip significantly enhanced IgM (64%) and IgG2a (50%) antibody production in mice treated with LPS mitogen. The same dose had no effect on IgM-specific response, whereas the 5 mg/kg treatment caused a statiscally significant reduction of anti-SRBC IgM-specific antibodies (82%). The aqueous extract of A. tenella (50 mg/kg) increased the life span (from 16 ± 1 to 25 ± 1 days) and decreased the number of viable tumor cells (59%) in mice with Ehrlich ascites carcinoma. The present findings are significant for the development of alternative, inexpensive and perhaps even safer strategies for cancer treatment.

In vitro and in vivo antitumor activity of crude extracts obtained from Brazilian Chromobacterium sp isolates

Natural products produced by microorganisms have been an important source of new substances and lead compounds for the pharmaceutical industry. Chromobacterium violaceum is a Gram-negative β-proteobacterium, abundant in water and soil in tropical and subtropical regions and it produces violacein, a pigment that has shown great pharmaceutical potential. Crude extracts of five Brazilian isolates of Chromobacterium sp (0.25, 2.5, 25, and 250 µg/mL) were evaluated in an in vitro antitumor activity assay with nine human tumor cells. Secondary metabolic profiles were analyzed by liquid chromatography and electrospray ionization mass spectrometry resulting in the identification of violacein in all extracts, whereas FK228 was detected only in EtCE 308 and EtCE 592 extracts. AcCE and EtCE 310 extracts showed selectivity for NCI/ADR-RES cells in the in vitro assay and were evaluated in vivo in the solid Ehrlich tumor model, resulting in 50.3 and 54.6% growth inhibition, respectively. The crude extracts of Chromobacterium sp isolates showed potential and selective antitumor activities for certain human tumor cells, making them a potential source of lead compounds. Furthermore, the results suggest that other compounds, in addition to violacein, deoxyviolacein and FK228, may be involved in the antitumor effect observed.

Antioxidant activity in vivo and in vitro of Halimeda incrassata aqueous extracts

The aim of the present paper was to provide the evidences for the antioxidant activity in Halimeda incrassata (Ellis) Lamouroux aqueous extracts obtained after simple water extraction of the fresh algae at room temperature (23°C). Previously in the literature, only antioxidant activity associated to carotenoids fractions of seaweeds has been reported. From different species of seaweeds, Halimeda incrassata aqueous extract exhibited the highest antioxidant activity on the inhibition of TBARS formed during the spontaneous lipid peroxidation of rat brain homogenates with an IC50 of 0.340mg.mL-1. Halimeda incrassata aqueous extract (0.5mg.mL-1), was also capable of decreasing the in vitro generation of hydrogen peroxide by two distinct metabolic pathways involving glutamic and malonic acids. Also, Halimeda incrassata (at doses of 50, 100 and 200mg.Kg-1) showed a neuroprotective effect in vivo on the gerbil model of bilateral carotid occlusion because of decreasing the locomotor and exploratory activity induced by ischemia. In summary, Halimeda incrassata aqueous extracts exhibit antioxidant properties in different in vitro as well as in vivo models which could be explained by the presence of several hydrosoluble compounds. Further studies on this way are necessary to elucidate the precise structure of these compounds. Low toxicity of most seaweeds to humans, but particularly of Halimeda genus may favor its use as functional food.

Phenols, flavonoids and antioxidant activity of aqueous and methanolic extracts of propolis (Apis mellifera L.) from Algarve, South Portugal

Propolis is a resinous substance collected by honeybees to seal honeycomb, which has been used in folk medicine due to its antimicrobial and antioxidant properties. In the present study, water and methanol were used to extract phenols and flavonoids from propolis collected in thirteen different areas in the Algarve region during the winter and spring. The ABTS•+, DPPH•, and O2•- scavenging capacity, and metal chelating activity were also evaluated in the propolis samples. Methanol was more effective than water in extracting total phenols (2.93-8.76 mg/mL) (0.93-2.81 mg/mL). Flavones and flavonols were also better extracted with methanol (1.28-2.76 mg/mL) than with water (0.031-0.019 mg/mL). The free radical scavenging activity, ABTS (IC50=0.006-0.036 mg/mL), DPPH (IC50=0.007-0.069 mg/mL) and superoxide (IC50=0.001-0.053 mg/mL), of the samples was also higher in methanolic extracts. The capacity for chelating metal ions was higher in aqueous extracts (41.11-82.35%) than in the methanolic ones (4.33-29.68%). Propolis from three locations of Algarve region were richer in phenols and had better capacity for scavenging free ABTS and DPPH radicals than the remaining samples. These places are part of a specific zone of Algarve known as Barrocal.

Developing novel one-step processes for obtaining food-grade O/W emulsions from pressurized fluid extracts: processes description, state of the art and perspectives

Abstract In this work, a novel on-line process for production of food-grade emulsions containing oily extracts, i.e. oil-in-water (O/W) emulsions, in only one step is presented. This process has been called ESFE, Emulsions from Supercritical Fluid Extraction. With this process, emulsions containing supercritical fluid extracts can be obtained directly from plant materials. The aim in the conception of this process is to propose a new rapid way to obtain emulsions from supercritical fluid extracts. Nowadays the conventional emulsion formulation method is a two-step procedure, i.e. first supercritical fluid extraction for obtaining an extract; secondly emulsion formulation using another device. Other variation of the process was tested and successfully validated originating a new acronymed process: EPFE (Emulsions from Pressurized Fluid Extractions). Both processes exploit the supercritical CO2-essential oils miscibility, in addition, EPFE process exploits the emulsification properties of saponin-rich pressurized aqueous plant extracts. The feasibility of this latter process was demonstrated using Pfaffia glomerata roots as source of saponin-rich extract, water as extracting solvent and clove essential oil, directly extracted using supercritical CO2, as a model dispersed phase. In addition, examples of pressurized fluid-based coupled processes applied for adding value to food bioactive compounds developed in the past five years are reviewed.

Morphological, physical and pedogenetic attributes related to water yield in small watersheds in Guarapari/ES, Brazil

Soil characteristics related to the genesis, land use and management are important factors in water dynamics in watersheds. This study evaluated physical, morphological and pedogenetic attributes related to water yield potential in small watersheds in Guarapari, ES, Brazil. The following representative profiles were selected, morphologically described and sampled in area of Atlantic Forest domain: Lithic Udifolists, Oxyaquic Udifluventes, Typic Paleudults, Typic Hapludults, Typic Hapludox, Oxic Dystrudepts and Typic Endoaquents. Samples were collected in the soil profiles for physical analysis. Measurements of field-saturated hydraulic conductivity and soil penetration resistance were perfomed in some profiles, which were under different uses. The Endoaquents of Limão Creek can be considered efficient as temporary water reservoirs. However, the use of artificial drainage tends to reduce this effect. Differential erosion was detected by the sand texture on the surface of the Typic Paleudults due to the low degree of clay flocculation, slope, high resistance to the penetration and low hydraulic conductivity of the Bt horizon, making it necessary to adopt soil management practices to increase the water infiltration. Under pasture, mainly in the cattle trails where the trampling is more intense, there was high resistance to penetration in the superficial layers of the Typic Hapludults. The Typic Hapludox have the greatest potential for water yield in the small watersheds because of its greater extent in the headwaters and their morphological and physical characteristics, which can result in increased aquifer recharge.

Effect of cattle manure on sunflower production and water use in two types of soil

The aim of the present study was to evaluate water consumption, use efficiency and yield components of sunflower variety Embrapa 122 V/2000 cultivated in two types of soil (Fluvissol and Haplic Luvisol) subjected to increasing doses of cattle manure. The experiment was carried out in a greenhouse at Universidade Estadual da Paraíba. The experimental design was completely randomized in a factorial scheme. The irrigation was performed every other day, replacing the water absorbed by the plants. The water consumption and the use efficiency were evaluated, being the use efficiency determined by the ratio of the total dry mass of sunflower and the amount of water used to produce it in each treatment. Plants were harvested at 95 days after sowing when the following parameters were evaluated: number of seeds per plant, weight of seeds per plant, weight of 1000 seeds and the outer diameter of the capitulum (head). The results showed that the sunflower was positively affected by cattle manure application, increasing the production components and the water use efficiency, regardless of the type of soil. Excepting for the 1000 seeds weight and the water use efficiency, the type of soil affected significantly the water use, the number and weight of seeds per plant. The plants cultivated in Haplic Luvisol had a better performance.

Nitrate leaching through climatologic water balance in a fertigated coffee plantation

Nitrate losses from soil profiles by leaching should preferentially be monitored during high rainfall events and during irrigation when fertilizer nitrogen applications are elevated. Using a climatologic water balance, based on the models of Thornthwaite and Penman Monteith for potential evapotranspiration, drainage soil water fluxes below the root zone were estimated in a fertigated coffee crop. Soil solution extraction at the depth of 1 m allowed the calculation of nitrate leaching. The average nitrate concentration in soil solution for plots that received nitrogen by fertigation at a rate of 400 kg ha-1, was 5.42 mg L-1, surpassing the limit of the Brazilian legislation of 10.0 mg L-1, only during one month. For plots receiving 800 kg ha-1 of nitrogen, the average was 25.01 mg L-1, 2.5 times higher than the above-mentioned limit. This information indicates that nitrogen rates higher than 400 kg ha-1 are potentially polluting the ground water. Yearly nitrate amounts of leaching were 24.2 and 153.0 kg ha-1 for the nitrogen rates of 400 and 800 kg ha-1, respectively. The six times higher loss indicates a cost/benefit problem for coffee fertigations above 400 kg ha-1.