Vânia Maria Maciel Melo

Larvicidal activity of the essential oil from Lippia sidoides cham. against Aedes aegypti linn

The aim of this work was to study the larvicidal activity of Lippia sidoides essential oil against Aedes aegypti larvae. The essential oil and its hydrolate (saturated solution of essential oil in water) were obtained by vapor extraction and their chemical composition determined by GL-chromatography coupled to mass spectroscopy. Bioassays were run with the essential oil, pure and diluted hydrolate and with their main constituents thymol and carvacrol. The results obtained showed that L. sidoides essential oil and its hydrolate have larvicidal action against the mosquito A. aegypti, causing an almost instantaneous mortality. Thymol, an alkylated phenol derivative and one of the major components of L. sidoides essential oil, was identified as the active principle responsible for the larvicidal action, causing 100% larval mortality at the lowest tested concentration of 0.017% (w/v). These results suggest that the essential oil of L. sidoides is promising as larvicide against A. aegypti and could be useful in the search of newer, more selective, and biodegradable larvicidal natural compounds to be used in official combat programs and at home.

Antibacterial activity of extracts of six macroalgae from the northeastern brazilian coast

Hexane, chloroform and ethanol extracts of six marine macroalgae (Rhodophyta and Chlorophyta) from North Ceara coast (Northeast Brazil) were evaluated for antibacterial activity by the single disk method. Best results were shown by the hexane extracts of Amansia multifida against enteric Gram-negative strains such as Enterobacter aerogenes, Klebsiella pneumoniae, Pseudomonas aeruginosa, Salmonella typhi, S. cholerae-suis, Serratia marcescens, Vibrio cholerae and the Gram-positive bacteria Bacillus subtilis and Staphylococcus aureus.

Kinetic study of biosurfactant production by Bacillus subtilis LAMI005 grown in clarified cashew apple juice

In this work a low cost medium for the production of a biosurfactant by Bacillus subtilis LAMI005 and the kinetics of surfactin production considering the effect of initial substrate concentration were investigated. First, cashew apple juice supplementation for optimal production of biosurfactant by B. subtilis LAMI005 was studied. The medium formulated with clarified cashew apple juice and distilled water, supplemented with 1.0 g/L of (NH(4))(2)SO(4), proved to be the best among the nutrients evaluated. The crude biosurfactant had the ability to decrease the surface tension of water to 30 dyne/cm, with a critical micelle concentration (CMC) of 63.0 mg/L. Emulsification experiments indicated that this biosurfactant effectively emulsified kerosene (IE(24)=67%) and soybean oil (IE(24)=64%). Furthermore, the emulsion stability was always very high. It was shown by biochemical analysis, IR spectra, that there is no qualitative differences in the composition of the crude biosurfactant from a standard sample of surfactin from B. subtilis.

Larvicidal activity of the water extract of Moringa oleifera seeds against Aedes aegypti and its toxicity upon laboratory animals

In this work, biological effects of the water extract of Moringa oleifera seeds (WEMOS) were assessed on eggs and 3rd instar larvae of Aedes aegypti and on its toxicity upon laboratory animals (Daphnia magna, mice and rats). Crude WEMOS showed a LC50 value of 1260microg/mL, causing 99.2 +/- 2.9% larvae mortality within 24 h at 5200microg/mL, though this larvicidal activity has been lost completely at 80 masculineC/10 min. WEMOS did not demonstrate capacity to prevent egg hatching. After extensive dialyses of the crude WEMOS into watersoluble dialyzable (DF) and nondyalizable (NDF) fractions, only DF maintained its efficacy to kill larvae. Acute toxicity evaluations on daphnids (EC50 of 188.7microg/mL) and mice (LD50 of 446.5 mg/kg body weight) pointed out to low toxicity. Despite the thymus hypertrophy, WEMOS revealed to be harmless in orally and subacutelytreated rats. In conclusion, WEMOS has thermostable bioactive compounds against Ae. aegypti larvae with apparent molecular mass lower than 12 kDa and moderately toxic potential.

Nutritional study of two Brazilian soybean (Glycine max) cultivars differing in the contents of antinutritional and toxic proteins

The research was conducted with two different recently released Brazilian soybean cultivars (Rio Balsas and Bays) to evaluate whether there is any correlation between the different levels of antinutritional and/or toxic proteins in the cultivars and their nutritive value as sources of protein for monogastric animals (rats). Furthermore, it is discussed, for the first time, the role of the dietary soyatoxin on the performance of rats fed on diets containing soyatoxin-rich (cv. Bays) and soyatoxin-free (cv. Rio Balsas) soybean cultivars. Feeding rats with diets containing raw soybean cultivars showed a lower growth rate, net protein utilization and digestibility, a much higher dry matter and nitrogen excretion and macroscopic alterations in internal organs when compared to rats fed on egg-white protein. The nutritional parameters measured for the diet based on raw Bays cultivar were poorer than those of the diet prepared with Rio Balsas. In the raw soybeans, trypsin inhibitor and lectin, and urease to a lesser extent, significantly affected at different fashion the soybean protein utilization. Heating treatment of the Bays seeds increased the growth rate, NPU, in vivo protein digestibility and practically eliminated or attenuated all the organ alterations observed. This study might be helpful in the choice of safe and nutritious soybean cultivars.

Purification and physicochemical characterization of a cotyledonary lectin from Luetzelburgia auriculata.

A lectin was purified from the cotyledons of Luetzelburgia auriculata (Fr. All) Ducke by affinity chromatography on agarose-N-acetyl-D-galactosamine. The lectin is a potent agglutinin for rabbit erythrocytes, reacts with human red cells, but is inactive against cow, sheep, and goat erythrocytes. Hemagglutination of rabbit erythrocytes was inhibited by either 0.39 mM N-acetyl-neuraminic acid or N-acetyl-D-galactosamin, 12.5 mM D-lactose or D-melibiose, 50 mM D-galactose or raffinose. Its hemagglutinating activity was lost at 80 degrees C, 5 min, and the activation energy required for denaturation was 104.75 kJ mol(-1). Chromatography on Sephadex G-100, at pH 7.6, showed that at this hydrogenic ionic concentration the native lectin was a homotetramer (123.5 kDa). By denaturing SDS-PAGE, LAA seemed to be composed of a mixture of 29 and 15 kDa polypeptide subunits. At acidic and basic pHs it assumed different conformations, as demonstrated by exclusion chromatography on Superdex 200 HR 10/30. The N-terminal sequence of the 29 kDa band was SEVVSFSFTKFNPNQKDII and the 15 kDa band contained a mixture of SEVVSFSFTKFNPNQKDII and KFNQIVAVEEDTDXESQPQ sequences, indicating that these bands may represent full-length and its endogenous fragments, respectively. The lectin is a glycoprotein having 3.2% neutral carbohydrate, with a pI of 5.8, containing high levels of Asp+Asn and Glu+Gln and hydroxy amino acids, and low amount or absence of sulfur amino acids. Its absorption spectrum showed a maximum at 280 nm and a epsilon (1%) x (1cm) of 5.2. Its CD spectrum was characterized by minima near 228 nm, maxima near 196 nm and a negative to positive crossover at 210 nm. The secondary structure content was 6% alpha-helix, 8% parallel beta-sheet, 38% antiparallel beta-sheet, 17% beta-turn, 31% unordered and others contribution, and 1% RMS (root mean square). In the fluorescence spectroscopy, excitation of the lectin solution at 280 nm gave an emission spectrum in the 285-445 nm range. The wavelength maximum emission was in 334.5 nm, typical for tryptophan residues buried inside the protein.

Purification of a novel antibacterial and haemagglutinating protein from the purple gland of the sea hare, Aplysia dactylomela Rang, 1828

Physicochemical characterisation and antibacterial and haemagglutinating properties of a new protein isolated from purple fluid of the Aplysia dactylomela are reported. The purification procedure consisted basically of ammonium sulphate fractionation, ion exchange, exclusion molecular and hydrophobic interaction chromatography. The highly purified protein, designated dactylomelin-P, is a single chain protein of 60,000 Da by SDS-polyacrylamide gel electrophoresis and 56,200 Da by gel filtration on calibrated Superose column at pH 7.5 and contains less than 0.05% of its weight in neutral carbohydrates. Dactylomelin-P has two biological activities, antibacterial and haemagglutinating. The antibacterial action is bacteriostatic but not bactericidal. The haemagglutinating activity is preferentially against rabbit erythrocytes. The glycoprotein fetuin was able to abolish the haemagglutinating activity but not the antibacterial one even when used at concentrations 10 fold higher. This is the first time that a chimeroprotein is described in the purple fluid of sea hares, which may be involved in the chemical defence mechanism of these organisms.

New approach for petroleum hydrocarbon degradation using bacterial spores entrapped in chitosan beads.

Spores of Bacillus subtilis LAMI008 were entrapped in 3-mm chitosan beads and cross-linked with 0.3% glutaraldehyde for n-hexadecane biodegradation and biosurfactant recovery. When exposed to nutrients, the spores generated vegetative cells without morphological alterations as revealed by atomic force microscopy. The entrapped cells degraded almost 100% of 1% of n-hexadecane in medium supplemented with 1% glucose and produce biosurfactant within 48 h, as well as free cells. The number of viable cells inside the beads was maintained throughout the n-hexadecane degradation process and the released biosurfactant was not used as a carbon source. Entrapment of bacterial spores in chitosan beads overcomes problems with stability, storage, and long term cell viability encountered with vegetative cells. This approach can potentially be utilized for biodegradation of complex compounds by entrapping spores of different species of bacteria.

Effect of proteins from the red seaweed Hypnea musciformis (Wulfen) Lamouroux on the growth of human pathogen yeasts

A protein fraction, rich in lectin, obtained from the red seaweed Hypnea musciformis by precipitation with ammonium sulfate (F40/70) was screened for chitinase and b-1,3-glucanase activity and assessed for antifungal potential against the human pathogen yeasts Candida albicans and C. guilliermondii. The F40/70 fraction showed chitinase and b-1,3-glucanase enzymes, with specific activities of 276.43 and 1880.7 Units.mg -1 protein, respectively. It was capable of inhibiting the growth of C. guilliermondii at the concentrations of 45, 100 and 450 µg protein.ml -1 but it showed only a discrete inhibition against C. albicans irrespective of the tested concentrations. The inhibitory action was shown to be fungistatic and the presence of the glycoprotein fetuin, for which the lectin in the fraction had affinity, abolished the antifungal action. The complete growth recovery following fetuin treatment indicated that chitinase and b-1,3-glucanase were not involved in the growth inhibition of these yeasts.

Antimicrobial effect of a crude sulfated polysaccharide from the red seaweed Gracilaria ornata

The aim of this study was to determine the yield, chemical composition, specific rotation (SR), infrared (IR) spectroscopy and the effect on bacterial growth of a crude sulfated polysaccharide (SP) from the red marine alga G. ornata (Go). Go-1 (25°C), Go-2 (80°C), and Go-3 (80°C) were sequentially extracted and yielded 9.2%. The contents of sulfate (5.88-10.3%) and proteins (0.1-3.7%) were small. The values of SR were [µ]D20°f -19.0, -51.0, and -56.5, respectively. IR spectrums showed the presence of galactose-4 sulfate and absence of 3,6-anydrogalactose-2 sulfate, galactose-6 sulfate and galactose-2 sulfate. SR and IR techniques confirmed SPs. Go-3 was tested on the growth of bacteria (Bacillus subtilis, Staphylococcus aureus, Enterobacter aerogens, Escherichia coli, Pseudomonas aeruginosa, Salmonela choleraesuis and Salmonela typhi), but only E. coli was inhibited.