Effect of different substrates on composting of poultry litter

The objective was to evaluate the differences between distinct types of litter material and their combinations in the dynamics of degradation on the organic matter fractions and the quality of the final compound. The treatments were established according to material used as substrate for broiler litter: treatment 1 - rice husks; 2 - sugar cane bagasse; 3 - wood shavings; 4 - wood shavings + sugar cane bagasse; 5 - rice husks + sugar cane bagasse; and 6 - Napier grass. The following variables were monitored: temperature, levels of total solids (TS), volatile solids (VS), mass and volume of the pile, fibrous fraction, and levels and reductions of N, P and K during the process. Piles formed with Napier grass and sugar cane bagasse presented the highest average temperatures during composting. The greater average reductions in TS and VS were attained in piles with sugar cane bagasse (68.12 and 73.07%, for TS and VS, respectively). The reductions of greatest volume occurred in piles with sugar cane bagasse (52.08%), followed by Napier grass (50.56%). Poultry litters composed of rice husks and wood shavings presented 13.21 and 10.23% of lignin, respectively, which contributed to the lower degradation of fibrous fraction and degradability. Substrates with lower lignin content were those with greatest organic matter degradation rate and had reduced losses of N levels during the process. Composting performance is affected by the initial substrate used to compose the poultry litter.

Thermostable conidial and mycelial alkaline phosphatases from the thermophilic fungus Scytalidium thermophilum

An extracellular (conidial) and an intracellular (mycelial) alkaline phosphatase from the thermophilic fungus Scytalidium thermophilum were purified by DEAE-cellulose and Concanavalin A-Sepharose chromatography. These enzymes showed allosteric behavior either in the presence or absence of MgCl2, BaCl2, CuCl2, and ZnCl2. All of these ions increased the maximal velocity of both enzymes. The molecular masses of the conidial and mycelial enzymes, estimated by gel filtration, were 162 and 132 kDa, respectively. Both proteins migrated on SDS-PAGE as a single polypeptide of 63 and 58.5 kDa, respectively, suggesting that these enzymes were dimers of identical subunits. The best substrate for the conidial and mycelial phosphatases was p-nitrophenylphosphate, but,beta -glycerophosphate and other phosphorylated compounds also served as substrates. The optimum pH for the conidial and mycelial alkaline phosphatases was 10.0 and 9.5 in the presence of AMPOL buffer, and their carbohydrate contents were about 54% and 63%, respectively. The optimum temperature was 70-75 degreesC for both activities. The enzymes were fully stable up to 1 h at 60 degreesC. These and other properties suggested that the alkaline phosphatases of S. thermophilum might be suitable for biotechnological applications.

Efeitos da amonização sobre a ocorrência de fungos, composição química e digestibilidade in vitro de fenos de grama Seda (Cynodon dactylon (L.) Pers.)

The experiment was conducted with grama seda (Cynodon dactylon (L.) Pers.) hays stored with a low moisture content (12-15%) and without chemical treatment, and hays stored with a high moisture content (20-25%) and treated with anhydrous ammonia (NH3) al 0.5 and 1.0% of DM, and urea at 0.9 and 1.8% of DM. At 65 days after treatment (AT) under a plastic cover, the bales were opened and samples were taken at 3, 15 and 30 days to determine the chemical composition and in vitro digestibility (IVDMD) of the hays. For the identification of fungi, samples were taken at 0, 15 and 30 days AT. The data were analyzed according to a split-plot design with the effects of the chemical treatments studied in the main plot and the effects of the periods of post-treatment studied in the sub-plots, Fourteen genera of fungi were observed in the hays, not treated and treated with NH3 and urea, with a higher occurrence of Cladosporium, Curvularia, Aspergillus, and Penicillium. Treatment with anhydrous ammonia and 1.8% urea controlled the occurrence of Aspergillus; however, Penicillium decreased in hays treated with ammonia 30 days AT. Ammoniation did not influence the contents of ADF, cellulose and lignin in the hays, but NDF and hemicellulose decreased with the use of ammonia 30 days AT. The CP contents and the IVDMD increased with ammoniation. The CP contents decreased in hays treated with NH3 as days AT increase, while hays treated with urea did not change.

Xylanases from fungi: properties and industrial applications

Xylan is the principal type of hemicellulose. It is a linear polymer of beta-D-xylopyranosyl units linked by (1-4) glycosidic bonds. In nature, the polysaccharide backbone may be added to 4-O-methyl-alpha-D-glucuronopyranosyl units, acetyl groups, alpha-L-arabinofuranosyl, etc., in variable proportions. An enzymatic complex is responsible for the hydrolysis of xylan, but the main enzymes involved are endo-1,4-beta-xylanase and beta-xylosidase. These enzymes are produced by fungi, bacteria, yeast, marine algae, protozoans, snails, crustaceans, insect, seeds, etc., but the principal commercial source is filamentous fungi. Recently, there has been much industrial interest in xylan and its hydrolytic enzymatic complex, as a supplement in animal feed, for the manufacture of bread, food and drinks, textiles, bleaching of cellulose pulp, ethanol and xylitol production. This review describes some properties of xylan and its metabolism, as well as the biochemical properties of xylanases and their commercial applications.

Purification and biochemical characterization of an endoxylanase from Aspergillus versicolor

An endoxylanase (beta-1,4-xylan xylanohydrolase, EC 3.2.1.8) was purified from the culture filtrate of a strain of Aspergillus versicolor grown on oat wheat. The enzyme was purified to homogeneity by chromatography on DEAE-cellulose and Sephadex G-75. The purified enzyme was a monomer of molecular mass estimated to be 19 kDa by SDS-PAGE and gel filtration. The enzyme was glycoprotein with 71% carbohydrate content and exhibited a pI of 5.4. The purified xylanase was specific for xylan hydrolysis. The enzyme had a K-m of 6.5 mg ml(-1) and a V-max of 1440 U (mg protein)(-1). (C) 1998 Federation of European Microbiological Societies. Published by Elsevier B.V. B.V. All rights reserved.

PURIFICATION AND PROPERTIES OF SHIKIMATE DEHYDROGENASE FROM CUCUMBER (CUCUMIS-SATIVUS L)

Shikimate dehydrogenase (SDH, EC 1.1.1.25) extracted from cucumber pulp (Cucumis sativus L.) was purified 7-fold by precipitation with ammonium sulfate and elution from columns of Sephadex G-25, DEAE-cellulose, and hydroxyapatite. Two activity bands were detected on polyacrylamide gel electrophoresis at the last purification step. pH optimum was 8.7, and molecular weight of 45 000 was estimated on a Sephadex G-100 column. SDH was inhibited competitively by protocatechuic acid with a K(i) value of 2 x 10-4 M. K(m) values of 6 x 10-5 and 1 x 10-5 M were determined for shikimic acid and NADP+, respectively. The enzyme was completely inhibited by HgCl2 and p-(chloromercuri)benzoate (PCMB). NaCl and KCl showed partial protection against inhibition by PCMB. Heat inactivation between 50 and 55-degrees-C was biphasic, and the enzyme was completely inactivated after 10 min at 60-degrees-C. Incubation of SDH with either NADP+ or shikimic acid protected the enzyme against heat inactivation.

Identification of polycyclic aromatic hydrocarbons and methoxylated phenols in wood smoke emitted during production of charcoal

More than 130 organic substances in dichloromethane-methanol (4: 1) extracts of particulate matter and the gaseous phase from wood burning for the production of charcoal have been identified by capillary gas chromatography coupled with low-resolution mass spectrometry (GC-MS), use of GC retention indices, and comparison with authentic standards. Many of the substances identified are methoxyphenols (derivatives of syringol and guaiacol), polycyclic aromatic hydrocarbons (PAH), oxidized PAH (oxy-PAH), and levoglucosan, the last being a monosoccharide derivative from the thermal breakdown of cellulose. The amount of unsubstituted PAH was greater than that of methyl- and dimethyl-substituted homologs.

Influence of temperature on the properties of the xylanolytic enzymes of the thermotolerant fungus Aspergillus phoenicis

This study reports on the effects of growth temperature on the secretion and some properties of the xylanase and beta-xylosidase activities produced by a thermotolerant Aspergillus phoenicis. Marked differences were observed when the organism was grown on xylan-supplemented medium at 25 degreesC or 42 degreesC. Production of xylanolytic enzymes reached maximum levels after 72 h of growth at 42 degreesC; and levels were three- to five-fold higher than at 25 degreesC. Secretion of xylanase and beta-xylosidase was also strongly stimulated at the higher temperature. The optimal temperature was 85 degreesC for extracellular and 90 degreesC for intracellular beta-xylosidase activity, independent of the growth temperature. The optimum temperature for extracellular xylanase increased from 50 degreesC to 55 degreesC when the fungus was cultivated at 42 degreesC. At the higher temperature, the xylanolytic enzymes produced by A. phoenicis showed increased thermo stability, with changes in the profiles of pH optima. The chromatographic profiles were distinct when samples obtained from cultures grown at different temperatures were eluted from DEAE-cellulose and Biogel P-60 columns.

Efeito de diferentes relações volumoso: concentrado no consumo, digestibilidade aparente e partição da digestão da dieta de bovinos

Three steers equipped with ruminal and duodenal cannulas were fed roughage:concentrate ratios 80:20, 60:40 and 40:60 in order to study intake and apparent, rumen and post-rumen digestibilities. The roughage was ''coast cross'' (Cynodon dactylon) hay (5.67% CP and 83.30% NDF). Undigestible neutral detergent fiber (NDF) was used as dry matter (DM) flow marker. DM intake means were 77.99, 91.03 and 91.81g DM/kg BW0.75, for the 20, 40 and 60% concentrate diets, respectively. DM intake for the 20% diet was statistically (P < 0.05) different from the other two diets. Apparent digestion coefficient (%) of DM (50.48, 57.32 and 61.33), organic matter (OM) (52.03, 58.91 and 62.76) and gross energy (GE) (48.95, 56.40 and 60.00) increased significantly with the increase in concentrate ratio of the diets. For the following components the apparent digestion coefficients were not statistically different: NDF (44.54, 45.28 and 42.53), ADF (40.69 44.39 and 43.60), cellulose (51.54, 54.34 and 52.03), hemicellulose (49.63, 46.78 and 39.18) and starch (86.59, 91.89 and 93.21). DM, OM, NDF, ATF, cellulose and starch ruminal and post-ruminal digestibilities were not statistically different. But the ruminal digestibilities of hemicellulose (94.81, 90.26 and 85.99) and EG (93.85, 83.30 and 78.77) decreased significantly as the concentrate ratio of the diets increased. The post-ruminal digestibility of hemicellulose (5.19, 9.74 and 14.03%) and GE (6.12, 16.20 and 21.23%) increased as the concentrate ratio of the diets increased.

Measurements of ambient ozone using indigo blue-coated filters

Ozone monitoring techniques utilize expensive instruments that are often large and heavy. These instruments are not easy to handle in the field, and their size also limits some sampling schemes, principally for indoor ozone determination. We have developed a lightweight, inexpensive, and sensitive method that offers flexibility to undertake measurements of ambient ozone in many environments, both indoor and outdoor. The method is based on the reaction of ozone with indigo blue dye. The indigo molecule contains 1 carbon double bond (C = C) that reacts with ozone and results in nearly colorless reaction products. During sample collection, 2 cellulose filters coated with 40 mu L of 1.0 x 10(-3) M indigo blue were used. The determinations were done spectrophotometrically at 250 and 600 nm. The analytical parameters studied were sampling time and flow rate. Analytical curves were constructed with concentrations ranging from 37 to 123 parts per billion by volume (ppbv) of standard ozone, at 0.4 L/min and 15 min sampling time. The detection limits achieved were 6 and 9 ppbv, respectively, at 250 and 600 nm. Considering interferences, measurements made at 250 nm gave more reliable and specific values for ozone.

ISOLATION AND IN-VITRO HYDROLYSIS OF GLOBULIN G1 FROM LENTILS (LENS-CULINARIS, MEDIK)

The major globulin fraction from lentil seeds was investigated with respect td in vitro hydrolysis by trypsin and chymotrypsin. Globulin was isolated by a NaCl-ascorbate extraction procedure and purified by DEAE-cellulose chromatography and gelfiltration chromatography on Sepharose CL-6B. The purity and identification of the protein were performed by PAGE. The native globulin, with a molecular weight of 375 kD, was resolved by SDS-PAGE into twelve polypeptides with molecular weights ranging from 61 to 14.5 kD. Native and heated globulin GI was hydrolyzed with trypsin and chymotrypsin. SDS-PAGE indicated that native globulin was more resistant to digestion than heated protein. Amino acid analysis of the major globulin revealed that glutamic acid was present in the largest concentration, followed by aspartic acid, arginine and leucine. As is also the case for other legumin-like globulins, lentil GI was deficient in sulfur-containing amino acids.

Partial purification and some properties of a T2 ribonuclease from Aspergillus flavipes

A ribonuclease was partially purified from the culture medium of Aspergillus flavipes (IZ:1501), after 96 h of cultivation by chromatography on DEAE-cellulose and Sephadex G100 columns. The molecular weight of the RNase was estimated to be 40 kD by gel filtration using Sephadex G100, and the optimum pH and temperature were 4.0 and 50-55 degrees C, respectively. Catalytic activity was inhibited by Zn+2, Fe+3, Hg+2 and Ag+ ions. The enzyme did not show an exact base specificity and produced four kinds of 3'-nucleotides from yeast RNA.

Valor nutritivo do feno de Brachiaria decumbens Stapf cv. Basilisk submetido a tratamento com amônia anidra ou uréia

The effects of the ammoniation of Brachiaria decumbens hay was evaluated. The hay bales were distributed into a complete randomized block design, with four replications and they were submitted to the treatments: untreated or treated with anhydrous ammonia (NH3)(2,0 and 3,0% of the DM) or with urea (3,6 and 5,4% of the DM). All the hays bales remained under plastic cover for 45 days. After three days of aeration, samples were collected for the determination of the chemical composition, nitrogenous compounds fraction and the in vitro dry matter (IVDDM) and organic matter (IVDOM) digestibility. In the metabolic study, Saanen goats breed was used in a 5x5 Latin squares design, where the apparent digestibility, the voluntary intake and the nutritive value index were evaluated. The ammoniation increased the contents of the total N, N ammonia (N-NH3) and non-protein N, with high effect on the levels of 3,0% of NH3 and 5,4% of urea. There were no differences between the level of 3,0% of NH3 and 5,4% of urea for the total N, N-NH3 and NPN. However, the treatment with 3,0% of NH3 allowed a larger fixation of N in ADIN and NDIN forms. The ammoniation increased the IVDMD and IVDMO and reduced the contents of neutral detergent fiber (NDF), hemicellulose, acid detergent fiber (ADF) and lignin, but it did not alter the cellulose and gross energy contents. The ammoniation increased the DM, OM, CP, NDF, ADF, hemicellulose, cellulose and gross energy apparent digestibility and as well as the voluntary intake of DM, digestible DM, digestible OM, digestible protein, digestible energy and the nutritive value index. The ammoniation increased the hay nutritive value index, but there were no differences between the levels of NH3 and urea.

Uso de indicadores para estimar fluxos de matéria seca ileal em eqüinos fistulados

This experiment was carried out with six half-bred Bretao-Campolinacolts with ileum fistulated to evaluate the difference in the ileum dry matter flow estimated by chromic oxide, cellulose, NDF, lignin and fecal lignin through the collection of ileum samples digesta at 28 h intervals, totalizing six samples per animal, starting at 10:00 a.m. The animals were fed ad libitum with the following diets: R1: clephantgrass, R2: elephantgrass plus ground corn, and R3: elephantgrass plus ground corn plus soybean meal. The data was statistically described, based on the coefficient of variation. The values of dry matter prececal digestion coefficients were, respectively, for diets 1, 2 and 3, at six schedules, for cellulose (-16.4; 21.4 and 6.6%), NDF(-34.7; 28.8 and -12.8%), to lignin (-51.5; -5.1 and -25.7%), in two schedules for cellulose (-13.4; 25.6 and 21.5%), fecal lignin (-37.1, 16.6 and -6.4%) and chromic oxide (-219.3, 36.4 and 9.5%). The coefficients of variation were, respectively, for the diets i, 2 and 3, at six schedules, for cellulose (148.3; 107.5 and 522.7%), NDF (95.4; 80.9 and 314.3%), lignin (210.2; 752.3 and 209.6%), at two schedules for cellulose(148.5; 80.7 and 70.0%), fecal lignin, (262.4; 177.9 and 723.5%) and chromic oxide (141.1; 158.9 and 473.4%). In diet i, the ileum dry matter now were over estimated for all markers, showing that chosen collection lime to estimate the flows were not adequate. Based on the coefficient of variation of the diets 2 and 3, the cellulose at two schedules was the most marker indicator to determine the ileum dry matter flow.

BIOSENSORS FOR GLUCOSE NEEDLE-SHAPED FOR IN-VIVO MONITORING

Different procedures for obtaining a needle biosensor for the determination of glucose to be inserted subcutaneously in vivo, have been compared. Platinum wires with a diameter of 75 mum, teflon-coated were inserted in hypodermic needles and fixed with a two-component epoxy resin. Using a dip-coating procedure, several layers were deposited on electrodes. The first coating was cellulose acetate, the second was immobilized glucose oxidase (GOD) mixed with bovine serum albumin (BSA) and glutaraldheyde, the third coating was a polyurethane coating obtained with commercially available products. A large number of electrodes have been tried and statistically evaluated but they seem to be affected by poor reproducibility evidenced by a large spreading in successive calibration curves. Then, the polyurethane coating has been replaced by a thin polycarbonate membrane salinized and fixed on the tip of the needle. Reproducible results were achieved and first results of in vivo measurements on rabbits are reported.