Cellulolytic activity of wild type and mutant Trichoderma pseudokoningii

Strains of Trichoderma pseudokoningii are promising objects for genetic studies and cellulase production. Auxotrophic mutants with deficiencies in the biosynthesis of aminoacids, nucleotides and vitamins (up to five markers) in addition to morphological aspects like conidial colour were obtained from two strains of double auxotrophic mutants using UV radiation. In order to compare the cellulolytic capabilities of the T. pseudokoningii (wild type strain), some of its mutants and T. reesei QM9414 we performed semiquantitative cellulase assays and quantitative determination of the enzymes exoglucanase and endoglucanase. The semiquantitative test showed that the strains with minimal mycelial growth rate were better producers. Both tests revealed that two of the studied mutants, TG3 and TG4 presented a yield higher than the wild type, reaching 30% more exoglucanase and 70% more endoglucanase. These results indicate that the wild type was improved for cellulase production. Highly significant values of correlation were found for exoglucanase and endoglucanase activities, suggesting that these enzymes may be co-regulated in T. pseudokoningii.

Invited review Giardia duodenalis: Inter-strain variability of proteins, antigens, proteases, isoenzymes and nucleic acids

Giardia duodenalis isolates from asymptomatic or symptomatic patients and from animals present similarities and differences in the protein composition, antigenic profile, pattern of proteases and isoenzymes, as well as in nucleic acids analysis. In the present overview, these differences and similarities are reviewed with emphasis in the host-parasite interplay and possible mechanisms of virulence of the protozoon.

Morphology of testis and epididymis in an ethanol-drinking rat strain (UChA and UChB)

The objective of the present study was to analyze the prospective alterations of the testis and epididymis in a defined strain of alcoholic rats in order to contribute to our understanding of the effects of chronic alcoholism on reproduction. The testis and epididymis of the animals were submitted to morphological analysis by macroscopy, light microscopy and electron microscopy and to morphometric analysis. The UCh rats showed atrophy of the epithelium and reduction of testis and epididymis weight, liver hypertrophy and fat infiltration and alterations of the hypothalamus-pituitary axis. Ethanol induces changes in the weight and in the epithelium of the testis and epididymis and in the hypothalamus-pituitary axis of the UCh rats.

Morphology of the ventral lobe of the prostate and seminal vesicles in an ethanol-drinking strain of rats (UChA and UChB)

Chronic alcoholism alters reproduction and therefore may be responsible for alterations of prostate and seminal vesicles, which are the subject of this analysis in UCh ethanol-drinking rats. The prostate and seminal vesicles of 20 animals were submitted to macroscopic, light microscopy, electron microscopy and morphometric analysis. The UCh rats showed atrophy of the epithelium and reduction of the weight of the prostate and seminal vesicle, liver hypertrophy and fat infiltration and alterations of the hypothalamus-pituitary axis. Ethanol induces changes in the weight and in the epithelium of prostate and seminal vesicles and hypothalamus-pituitary axis of UCh rats.

Study of crystallite size and strain as a function of morphological evolution in zinc oxide powder obtained from hydroxycarbonate precursor

In this work, zinc oxide samples were obtained from hydroxycarbonate by thermal decomposition at 300°C. Zinc hydroxycarbonate samples were produced by homogeneous precipitation over different periods of time. The method used to obtain zinc oxide produces different morphologies as a function of the precursor precipitation time. Among the obtained particle shapes were porous spherical aggregates, spherulitic needle aggregates, and single acicular particles. This work investigated spherulitic needle-aggregate formation and the correlation among morphology, domain size, and microstrain. Transmission electron microscopy data revealed that the acicular particles that form the spherulitic needle aggregates consist of nanometer crystallites. Apparent crystallite size and microstrain in the directions perpendicular to (h00), (h0l), (hk0), and (00l) planes were invariable as a function of precursor precipitation time. From the results, it was possible to conclude that the precursor precipitation period directly influenced the morphology of the zinc oxide but did not influence average crystallite size and microstrain for ZnO samples. Therefore, using this route, it was possible to prepare zinc oxide with different morphologies without microstructural alterations. © 2001 International Centre for Diffraction Data.

Production, characterization and properties of polysaccharide depolymerizing enzymes from a strain of Curvularia inaequalis

Xylanase, β-glucosidase, β-xylosidase, endoglucanase and polygalacturonase production from Curvularia inaequalis was carried out by means of solid-state and submerged fermentation using different carbon sources. β-Glucosidase, β-xylosidase, polygalacturonase and xylanase produced by the microorganisms were characterized. β-Glucosidase presented optimum activity at pH 5.5 whereas xylanase, polygalacturonase and β-xylosidase activities were optimal at pH 5.0. Maximal activity of β-glucosidase was determined at 60°C, β-xylosidase at 70°C, and polygalacturonase and xylanase at 55°C. These enzymes were stable at acidic to neutral pH and at 40-45°C. The crude enzyme solution was studied for the hydrolysis of agricultural residues.

Characterization of Rhizoctonia solani associated with soybean in Brazil

Rhizoctonia solani causes pre- and post-emergence damping-off, root and hypocotyl rot and foliar blight in soybean. Foliar blight has resulted in yield losses of 31-60% in north and northeast Brazil. The aim of this study was to characterize isolates of R. solani associated with soybean in Brazil. Among 73 Rhizoctonia isolates examined, six were binucleate and 67 were multinucleate. The multinucleate isolates were characterized according to hyphal anastomosis reaction, mycelial growth rate, thiamine requirement, sclerotia production, and RAPD molecular markers. Four isolates that caused hypocotyl rot belonged to AG-4 and using RAPD analysis they grouped together with the HGI subgroup. Another isolate that caused root and hypocotyl rots was thiamine auxotrophic, grew at 35 °C, and belonged to AG-2-2 IIIB. All 62 isolates that caused foliar blight belonged to AG-1 IA. RAPD analysis of R. solani AG-1 IA soybean isolates showed high genetic similarity to a tester strain of AG-1 IA, confirming their classification. The teleomorph of R. solani, Thanatephorus cucumeris was produced in vitro by one AG-1 IA isolate from soybean. The AG-4 and AG-2-2 IIIB isolates caused damping-off and root and hypocotyl rots of soybean seedlings cv. 'FT-Cristalina', under greenhouse conditions. The AG-2-2 IIIB isolate caused large lesions on the cortex tissue, that was distinct from the symptoms caused by AG-4 isolates. The AG-1 IA isolates caused foliar blight in adult soybean plants cv. 'Xingu' under the greenhouse and also in a detached-leaf assay.

Comparative experimental infection of Lacazia loboi in BALB/c and B10.A mice

Both hind foot pads of BALB/c and B10.A mice strains, were inoculated with a fungal suspension of Lacazia loboi obtained from a Jorge Lobo's disease patient. The suspension had 9 × 105 cells/ml and its viability index was 45%. The animals were sacrificed at different time periods varying from 24 h to 18 months after inoculation. The BALB/c mice developed an extensive granulomatous infiltrate, similar to the disease in humans, that progressively evolved. The number of fungal elements also increased as the disease progressed, and after the seventh month of inoculation, macroscopic changes of the foot pads were evident. Although the B10.A mice developed an exuberant granulomatous infiltrate, macroscopic changes were not detected. The number of fungal cells in the infected tissues increased in number, but they were lower then the numbers found in the BALB/c strain. The viability indexes were also lower for the B10.A strain. Considering the histopathological findings, the presence of macroscopic changes and the great amount of fungal cells in the infected tissues, the authors concluded that the BALB/c mice strain was more susceptible to L. loboi infection than the B10.A strain.

Morphology of the vas deferens in an ethanol-drinking strain of rats (UChA and UChB)

Chronic alcoholism alters reproduction and therefore may be responsible for alterations of vas deferens, which are the subject of this analysis in UCh ethanol-drinking rats. The proximal and distal segments of the vas deferens of 20 animals were submitted to macroscopic, light microscopy, electron microscopy and morphometric analysis. The UCh rats showed atrophy of the epithelium of the vas deferens and alterations of the hypothalamus-pituitary axis. Ethanol induces changes in the epithelium of the vas deferens and hypothalamus-pituitary axis of UCh rats.

Purification and characterization of xylanases from Aspergillus giganteus

A strain of Aspergillus giganteus cultivated in a medium with xylan produced two xylanases (xylanase I and II) which were purified to homogeneity. Their molar mass, estimated by SDS-PAGE, were 21 and 24 kDa, respectively. Both enzymes are glycoproteins with 50°C temperature optimum; optimum pH was 6.0-6.5 for xylanase I and 6.0 for xylanase II. At 50°C xylanase I exhibited higher thermostability than xylanase II. Hg2+, Cu 2+ and SDS were strong inhibitors, 1,4-dithiothreitol stimulated the reaction of both enzymes. Both xylanases are xylan-specific; kinetic parameters indicated higher efficiency in the hydrolysis of oat spelts xylan. In hydrolysis of this substrate, xylotriose, xylotetraose and larger xylooligosaccharides were released and hence the enzymes were classified as endoxylanases.

High sensitivity low cost capacitive strain gage

This paper describes a high senstivity low cost capacitive strain gage sensor. The theory, design, and sensor construction details are presented. It consists of eight capacitive sensors connected in two full bridges. The capacitive strain gage sensor structure was designed in order to produce high sensitivity and low dependence with temperature. By using a simple signal conditioning circuit constituted by a differential amplifier, a band-pass filter, and a precision rectifier the device can measure forces with resolution of 0.009 N and precision of 98.7%. It is rugged, presents linear response, and good repeatability. It presents sensitivity of 8.7 V/N and fall time of 12 ms.

Production of CGTase by a bacillus Alkalophilic CGII strain isolated from wastewater of a manioc flour industry

GCTase production by a new strain of Bacillus alkalophillc CGII isolated from Brazilian wastewater of manioc flour industry was examined. The growth medium used was composed by 1.5% starch, 1.5% nitrogen and 1% Na 2CO3. Higher activity was obtained with starch, maltodextrin and galactose. When glucose was added to the medium, no enzyme production was observed. High enzyme activity and growth were reached when aeration was increased (88.6 U/mL). The enzyme characterization showed an optimum pH and temperature 8.0 and 55°C for starch hydrolyses, respectively. Mg+ and Ca++ showed small activation; however, Hg + and Cu+ showed a strong enzyme inhibition.

Evaluation of serum- and animal protein-free media for the production of infectious bronchitis virus (M41) strain in a continuous cell line

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Antibiótico antifungico produzido por um estreptomiceto da região de Araraquara

With the significant increase in the incidence of invasive fungal infections during the last decade, mainly in patients with cancer, AIDS and other hospitalized patients who stay for long periods in intensive care units, there is an urgent need to screen for new antifungal agents possessing some advantages over known ones. This article reports a search in the field for a microorganism producing antibacterial and antifungal substances. Strains from soil samples collected in the region of Araraquara, Brazil, were isolated and analyzed for their antimicrobial potential against standard microorganisms (fungi Candida albicans and Aspergillus oryzae and bacteria Staphylococcus aureus and Escherichia coli). Out of the 64 strains isolated, 34 produced detectable antimicrobial activity. The streptomycete strain Ar4014 was chosen for further study, owing to its good antimicrobial activity against Candida albicans. Two of the fermentation media tested, 608-K and 602-B, were found to be best for the production and extraction of the antibiotic from Ar4014. After chromatographic separation of the crude extract on a silica column, the active fractions obtained showed UV-VIS absorption peaks characteristic of normal pentaenic antibiotics. The antibiotic was provisionally designated Ara 4014-75.

Pro- and anti-inflammatory cytokines produced by human monocytes challenged in vitro with Paracoccidioides brasiliensis

Monocytes and macrophages play a central role in innate and adaptive immune response against systemic fungal infections. Imbalances in suppressor or stimulatory cytokine secretion caused by these cells may influence disease development, microorganism death, and the nature of the adaptive immune response. This study analyzed the monocyte cytokine profiles of healthy individuals challenged with high and low virulent strains of P. brasiliensis and mRNA cytokine expression kinetics by reverse transcription polymerase chain reaction (RT-PCR). Peripheral blood monocytes from healthy volunteers were cultured in vitro with and without virulent (Pb18) or low virulence (Pb265) strains from P. brasiliensis viable yeast cells. Interleukin-1 beta (IL-1β), IL-6, IL-8, IL-10, tumor necrosis factor-alpha (TNF-α), and transforming growth factor-beta (TGF-β1) were measured in culture supernatants by enzyme immunoassay (ELISA), and mRNA cytokine expression was determined by RT-PCR at 0, 4, 8, 12, 18 and 48 hr. Both P. brasiliensis strains induced monocyte production of IL-1β, IL-6, IL-10 and TNF-α. Pb18 induced higher levels of IL-1β, IL-6, and IL-10 than Pb265. IL-8 and TGF-β1 levels were not significantly different from those cultured without stimulus. The mRNA cytokine expression was similar to supernatant cytokines measured by ELISA. In vitro monocyte challenge with virulent P. brasiliensis strain induces earlier and higher levels of pro- and anti-inflammatory cytokines than low virulence strain.