José Humberto de Queiroz

Low-temperature clean-up method for the determination of pyrethroids in milk using gas chromatography with electron capture detection

This paper describes a new gas-chromatography with electron capture detection (GC-ECD) method for determination of some pyrethroids in milk samples. The extraction of the pyrethroids was carried out by liquid-liquid extraction with clean-up by precipitation at low temperature, without additional stages for removal of fat interferences. The method was efficient with recoveries of 93.0+/-0.1% for cipermethrin and 84.0+/-0.3% for deltamethrin. The quantification limits were 0.75 microg L(-1) for both pyrethroids. The method was simple, of easy execution, and used only small quantities of organic solvent. After optimization and validation, the method was used for the determination of residues of the pyrethroids cipermethrin and deltamethrin in milk and in lactea drink commercialized in Viçosa (MG, Brazil). Some samples presented contamination with deltamethrin at levels below the maximum contamination limits established by the FAO.

Potencial da palha de cana-de-açúcar para produção de etanol

Sugarcane straw biomass accounts for 1/3 of the energy potential of sugarcane and represents a rich source of sugars. Studies have been intensified for the use of this biomass along with bagasse for the production of cellulosic ethanol. Development of this technological path will allow for taking full advantage of sugarcane, increasing ethanol production without expanding the area cultivated. However, in order for this technology to be viable certain challenges must be overcome, including establishment of appropriate conditions of pretreatment and hydrolysis of these materials for release of fermentable sugars.

Optimization and validation of liquid–liquid extraction with low temperature partitioning for determination of carbamates in water

Using a 2(3) experimental design, liquid-liquid extraction with low temperature partitioning (LLE-LTP) was optimized and validated for analysis of three carbamates (aldicarb, carbofuran and carbaryl) in water samples. In this method, 2.0 mL of sample is placed in contact with 4.0 mL of acetonitrile. After agitation, the sample is placed in a freezer for 3 h for phase separation. The organic extract is analyzed by high performance liquid chromatography with ultraviolet detection (HPLC-UV). For validation of the technique, the following figures of merit were evaluated: accuracy, precision, detection and quantification limits, linearity, sensibility and selectivity. Extraction recovery percentages of the carbamates aldicarb, carbofuran and carbaryl were 90%, 95% and 96%, respectively. Even though extremely low volumes of sample and solvent were used, the extraction method was selective and the detection and quantification limits were between 5.0 and 10.0 microg L(-1), and 17.0 and 33.0 microg L(-1), respectively.

Novel lactones from Aspergillus versicolor

Abstract Two novel aromatic lactones were isolated from Aspergillus versicolor fermentation broth. The structures were determined mainly by 1D and 2D NMR spectroscopy.

Ação ovicida do extrato bruto enzimático do fungo Pochonia chlamydosporia sobre ovos de Ancylostoma sp

INTRODUCTION Ancylostoma sp is a potentially zoonotic geohelminth. METHODS This study aimed to evaluate in vitro the action of crude enzyme extract of Pochonia chlamydosporia (VC4) on eggs of Ancylostoma sp in 2% water-agar and in fecal cultures. RESULTS The percentage reduction in Ancylostoma sp egg eclosion was 76.8% in Petri dishes of the treated group compared to the control group. CONCLUSIONS The crude enzyme extract of Pochonia chlamydosporia was effective at reducing Ancylostoma sp egg eclosion and can be used as biological control of this nematode.

Optimizing protease production from an isolate of the nematophagous fungus Duddingtonia flagrans using response surface methodology and its larvicidal activity on horse cyathostomins.

Protease production from Duddingtonia flagrans (isolate AC001) was optimized and the larvicidal activity of the enzymatic extract was evaluated on infective horse cyathostomin larvae (L3). Duddingtonia flagrans was grown in liquid medium with eight different variables: glucose, casein, bibasic potassium phosphate (K2HPO4), magnesium sulphate (MgSO4), zinc sulphate (ZnSO4), ferrous sulphate (FeSO4), copper sulphate (CuSO4) and temperature. The Plackett-Burman analysis showed a significant influence of MgSO4, CuSO4 and casein (P < 0.05) on protease production by D. flagrans in liquid medium. Central composite design indicated that the highest proteolytic activity was 39.56 U/ml as a function of the concentrations of casein (18.409 g/l), MgSO4 (0.10 g/l) and CuSO4 (0.50 mg/l). A significant difference (P < 0.01) was found for the larval number between the treated and control groups at the end of the experiment. A reduction of 95.46% in the number of free-living larvae was found in the treated group compared with the control. The results of this study suggest that protease production by D. flagrans (AC001) in liquid medium was optimized by MgSO4, CuSO4 and casein, showing that the optimized enzymatic extract exerted larvicidal activity on cyathostomins and therefore may contribute to large-scale industrial production.

Otimização do pré-tratamento hidrotérmico da palha de cana-de-açúcar visando à produção de etanol celulósico

The sugarcane industry has huge potential for biorefinery concept application, given its development in recent years. In this context, cane sugar straw has become an attractive raw material for biofuel production. This study aims to investigate the chemical composition of cane sugar straw from different regions of Brazil, and to optimize a hydrothermal pretreatment stage for cellulosic ethanol production. Results of chemical characterization of the cane sugar straw for the regions assessed indicated little influence of place on straw chemical composition. Hydrothermal pretreatment showed high efficiency in hemicellulose removal. Hydrothermal pretreatments operating with temperatures of 190 and 210 oC presented satisfactory results, reaching values close to 100% hydrolysis.

Interaction of the nematophagous fungus Duddingtonia flagrans on Amblyomma cajannense engorged females and enzymatic characterisation of its chitinase

Abstract The present work aimed to evaluate the production and the characterisation of a chitinase from nematophagous fungus Duddingtonia flagrans (AC001) and observe the interaction of this fungus on engorged females of Amblyomma cajennense under laboratory conditions. In assay A, the engorged females of A. cajennense were separated and immersed for 5 seconds in a fungal suspension of 106 conidia/ml of the fungus D. flagrans and placed in Petri dishes, in the dark. In assay B, wheat bran supplemented with 1% chitin and liquid minimal medium was used [K2HPO4 (5.0 g/l), MgSO4 (0.10 g/l), ZnSO4 (0.0050 g/l), FeSO4 (0.001 g/l) e CuSO4 (0.50 mg/l)], as a substrate for chitinase production. To demonstrate the presence of chitinase in the crude extract obtained after the enzymatic extraction, a purification process was developed using a specific adsorption technique. The results from assay A demonstrated the interaction of the D. flagrans conidia produced from chitin-agar on engorged females of A. cajennense. In the assay B, D. flagrans produced a chitinase successfully, with a high value for enzyme activity. The molecular mass of semi-purified enzyme was estimated at approximately 34 kDa. It was concluded that the fungus produced a chitinase and has some entomopathogenic activity, as demonstrated here for the first time; however, it is strongly suggested that further studies are needed to elucidate the molecular mechanism of infection of target organisms by this fungus.

Ovicidal action of a crude enzymatic extract of the fungus Pochonia chlamydosporia against cyathostomin eggs.

The aims of this study were to test the action of the fungal extract of Pochonia chlamydosporia (VC4) on the hatching of cyathostomin eggs plated in Petri dishes containing 2% water-agar (2% WA) and its enzymatic activity in fecal cultures, in two experimental assays (A and B). The fungus P. chlamydosporia (VC4) was cultured in Erlenmeyer flasks (250ml) containing 50ml of liquid minimal medium supplemented with 0.2% gelatin for production of the crude enzymatic extract. Approximately 1kg of fresh feces was collected directly from the rectum of crossbred horses naturally infected with cyathostomins. The fecal material was used to obtain eggs and prepare fecal cultures. For assay A, one thousand eggs were plated on 4.5cm diameter Petri dishes together with 5ml of VC4 fungal filtrate and incubated at 26 degrees C in the dark for 24h. The control group consisted of 1000 eggs in Petri dishes containing 10ml of distilled water, which were incubated under the same conditions. After 24h, the total number of cyathostomin larvae present in each plate of the treated and control groups was counted. For assay B, about 20g of feces were added with 10ml of fungal extract of P. chlamydosporia (VC4) and incubated at 26 degrees C for 8 days. Third stage larvae (L(3)) were recovered at the end of this period. Significant difference (p<0.01) was found for the number of larvae between the treated group and the control at end of assay A. A 72.8% reduction in the hatching of cyathostomin eggs was found in the plates of the treated group compared with the control group. At the end of 8 days, the fungal extract of P. chlamydosporia (VC4), in assay B, was effective in reducing the number of L(3) cyathostomins in the treated group by 67.0% compared with the control group. Significant difference (p<0.01) was found between the means of L(3) recovered from the treated group and the control group. The results of this work showed that crude enzymatic extract of P. chlamydosporia (VC4) was effective in reducing hatching of cyathostomin eggs and therefore could be used as a biological control agent of this nematode.

Pochonia chlamydosporia fungal activity in a solid medium and its crude extract against eggs of Ascaridia galli.

The present study aimed to evaluate the ovicidal activity (type 3 effect) of VC1 and VC4 isolates of Pochonia chlamydosporia in a solid medium and the action of a crude extract of P. chlamydosporia against eggs of Ascaridia galli. To evaluate ovicidal activity in culture medium, 1000 A. galli eggs were plated on Petri dishes containing 2% water-agar with grown fungal isolates (VC1 or VC4) and without fungus (control group) and were examined at 1, 3 and 5 days post-inoculation (assay A). Then, to test the action of crude extracts of P. chlamydosporia (VC1 or VC4), 500 eggs of A. galli were plated on Petri dishes of 4.5 cm diameter with 5 ml of fungal filtrate from each tested isolate. The control group consisted of 500 eggs of A. galli with 10 ml of distilled water on each Petri dish (assay B). Fungal isolates were effective (P < 0.01) at destroying these eggs, showing a type 3 effect at the studied intervals. On the other hand, the crude extract of isolates (VC1 or VC4) reduced the number of A. galli eggs in the treated group compared with the control group by 64.1% and 56.5%, respectively. The results of the present study show that P. chlamydosporia is effective at destroying eggs of A. galli and could therefore be used in the biological control of nematodes.