Isolation and growth of the symbiotic fungus of Atta capiguara (Hymenoptera: Formicidae)

This work's objectives were to isolate and evaluate the growth of the symbiotic fungus of Atta capiguara Gonçalves on artificial medium, under different pH and temperature conditions. Isolation was accomplished using the following media: Sabouraud, oat-agar, PDA, and PDA with the addition of extracts from the grasses Paspalum sp. Flügge and Hyparrhenia rufa (Nees) Stapf.. The medium used in the growth study was PDA with the addition of a Paspalum sp. (0.22%, w/v) extract at initial pH values of 4.5, 6.0, and 7.5. Mycelium disks were transferred to plates containing the culture medium. The plates were maintained at temperatures of 20, 23, and 26 ± 1°C. Mycelial radial growth evaluations were performed at 7, 14, 21, 28, and 35 days of incubation. Fungus isolation was obtained in all media studied. The highest radial means were obtained at initial pH values of 6.0 and 7.5 and temperatures of 23 and 26± 1°C. Greater plot losses occurred at the initial pH condition of 7.5. In general, A. capiguara fungi can be grown in the medium studied, at an initial pH of 6.0 and temperatures of 23 or 26± 1°C. Radial growth evaluations at 14 and 28 days of incubation can be recommended for substrate studies involving the symbiotic fungus.

Communiols E-H: New polyketide metabolites from the coprophilous fungus Podospora communis

Communiols E-H (1-4), four new polyketide-derived natural products containing furanocyclopentane, furanocyclopentene, cyclopentene, or γ-lactone moieties, have been isolated from two geographically distinct isolates of the coprophilous fungus Podospora communis. The structures of these compounds were determined by analysis of NMR and MS data. © 2005 American Chemical Society and American Society of Pharmacognosy.

Evaluation of DNA polymorphisms amplified by arbitrary primers (RAPD) as genetically associated elements to differentiate virulent and non-virulent Paracoccidioides brasiliensis isolates

Randomly amplified polymorphic DNA (RAPD) analysis of 35 Paracoccidioides brasiliensis isolates was carried out to evaluate the correlation of RAPD profiles with the virulence degree or the type of the clinical manifestations of human paracoccidioidomycosis. The dendrogram presented two main groups sharing 64% genetic similarity. Group A included two isolates from patients with chronic paracoccidioidomycosis; group B comprised the following isolates showing 65% similarity: two non-virulent, six attenuated, five virulent, eight from patients with chronic paracoccidioidomycosis and two from patients with acute paracoccidioidomycosis. The virulent Pb18 isolate and six attenuated or non-virulent samples derived from it were genetically indistinguishable (100% of similarity). Thus, in our study, RAPD patterns could not discriminate among 35 P. brasiliensis isolates according to their differences either in the degree of virulence or in the type of the clinical manifestation of this fungal infection. © 2002 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.

Molecular typing of IberoAmerican Cryptococcus neoformans isolates

A network was established to acquire basic knowledge of Cryptococcus neoformans in IberoAmerican countries. To this effect, 340 clinical, veterinary, and environmental isolates from Argentina, Brazil, Chile, Colombia, Mexico, Peru, Venezuela, Guatemala, and Spain were typed by using M13 polymerase chain reaction-fingerprinting and orotidine monophosphate pyrophosphorylase (URA5) gene restriction fragment length polymorphsm analysis with Hhal and Sau961 in a double digest. Both techniques grouped all isolates into eight previously established molecular types. The majority of the isolates, 68.2% (n=232), were VNI (var. grubii, serotype A), which accords with the fact that this variety causes most human cryptococcal infections worldwide. A smaller proportion, 5.6% (n=19), were VNII (var. grubii, serotype A); 4.1% (n=14), VNIII (AD hybrid), with 9 isolates having a polymorphism in the URA5 gene; 1.8% (n=6), VNIV (var. neoformans, serotype D); 3.5% (n=12), VGI; 6.2% (n=21), VGII; 9.1% (n=31), VGIII, and 1.5% (n=5) VGIV, with all four VG types containing var. gattii serotypes B and C isolates.

Epidemiologia de candidíase hospitalar: Importância da identificação específica

Nosocomial infections with Candida species are recognized as a significant cause of morbidity and mortality in both seriously ill immunocompetent and immunocompromised patients. Infections with Candida albicans and non-albicans Candida species have become a significant cause of infection in humans. Several of the more commonly Candida spp isolates are less susceptible to the antifungal drugs currentlly applied in clinical treatment, a factor that means significant difficulties for effective treatment. The modern mycology laboratory has an important role to play in several aspects relating to these organisms, including therapy, detection, identification and epidemiological analysis. In this study, we have provided an initial comparison of differences in species distribution among Candida isolates from four general hospitals of São Paulo,SP. Overall, 40 isolates of C. albicans, C. parapsilosis and C. tropicalis were isolated respectively in 35%, 50% and 15%, revealed a tendency of higher frequency of non-albicans species. The species distribution in patients with candidemia showed that the most commonly species were C. parapsilosis (45,4%), followed by C. albicans (36,4%) and C. tropicalis (18,2%); thus, we have an increase of non-albicans species. The three different species were include in 6, 3, and 4 different biotypes, respectively C. albicans, C. parapsilosis e C. tropicalis. This study emphasizes the importance of periodic evaluation of Candida species distribution especially in centers caring for patients at risk.

Isolamento de fungos produtores de enzimas pectinolíticas

Soil samples collected in the campus, UNESP, Araraquara, SP, were employed to isolate and characterize fungi strains with potential pectinolytic enzymes. These enzymes have arisen great interest due to its increasing application in the food industry. Two hundred forty six strains were isolated based on the appearance of colony on PDA medium, morphology (septate mycelia, nonseptate conidiophore, black conidia, and clublike spore-bearing head), after 48 h of growth at 30°C. Strains were selected in solid medium containing pectin citrus as sole carbon source and 0.5% rutenium red. The characterization of pectinolytic production was performed in solid culture and batch fermentation medium containing pectin citrus. The enzyme pectinolytic production was evaluated at 30°C, without agitation in 100 mL of medium containing 2% pectin citrus, 0.2% ammonium sulphate, 0.2% magnesium sulphate, and 0.05% potassium phosphate. The maximum pectinolytic activity (15U/mL) was observed in the medium after Aspergillus sp CFCF-0492 growth, while Aspergillus sp CFCF-CC1 showed the higher level of the final biomass. The pectinolytic activity is more preserved when the fungi-spores were maintained in agar-Czapeck medium.

Cryptococcosis outbreak in psittacine birds in Brazil

An outbreak of cryptococcosis occurred in a breeding aviary in São Paulo, Brazil. Seven psittacine birds (of species Charmosyna papou, Lorius lory, Trichoglossus goldiei, Psittacula krameri and Psittacus erithacus) died of disseminated cryptococcosis. Incoordination, progressive paralysis and difficulty in flying were seen in five birds, whereas superficial lesions coincident with respiratory alterations were seen in two birds. Encapsulated yeasts suggestive of Cryptococcus sp. were seen in faecal smears stained with India ink in two cases. Histological examination of the birds showed cryptococcal cells in various tissues, including the beak, choana, sinus, lungs, air sacs, heart, liver, spleen, kidneys, intestines and central nervous system. High titres of cryptococcal antigen were observed in the serum of an affected bird. In this case, titres increased during treatment and the bird eventually died. Yeasts were isolated from the nasal mass, faeces and liver of one bird. Cryptococcus neoformans var. gattii serovar B was identified based on biochemical, physiological and serological tests. These strains were resistant (minimum inhibitory concentration 64 μ g/ml) to fluconazole. This is the first report of C. neoformans var. gattii occurring in psittacine birds in Brazil. © 2004 ISHAM.

Microbiota fúngica de um prédio com laboratórios da região de Araraquara-SP

From September 2000 to January 2001, airborne fungi were isolated from the building of the Clinical Analyses laboratories, including its didactic and research rooms, in Araraquara São Paulo State, Brazil, by using Andersen, MAS-100® (MERCK) machine, with Sabouraud chloramphenicol medium. After 5 days of incubation at 25°C, the colonies of the fungi were counted, resulting in the identification of 21 taxa. Cladophialophora spp. was the most isolated in internal and external environments as well, followed by Penicillium spp. and Mycelia spp. In accordance with the resolution n° 9, January 2003 (ANVISA), fungi considered unacceptable were found in nine internal environments and one of these presented the amount of fungi above of the acceptable limit. Among the obtained fungi, at least 16 taxa were reported as opportunistic, nine were related to plant diseases and seven were associated to allergy problems.

Epidemiologia das dermatofitoses em instituições públicas da região de Araraquara - SP

Dermatophytosis on skin, hair and nails are the most common infectious process in the world. A total of 94 individuals from Public Institutions from the city of Araraquara - Sao Paulo/Brazil, with suspected of dermatophytic lesions were examined in order to determine the incidence and etiology of dermatophytosis. 105 specimens were collected from August to December of 2001 in the Mycology Laboratory of the Faculdade de Ciências Farmacêuticas. It was observed that 47 samples were positive for dermatophytes. Trichophyton rubrum was the prevalent specie (59.6%), followed by Microsporum canis (17%), T. tonsurans (10.6%), T. mentagrophytes (8.5%) and Epidermophyton floccosum (4.3%). T. rubrum was the most frequent in interdigital lesions (81.5%) and M. canis was the main dermatophyte involved in scalp lesions (58.3%). Therefore, it was observed a predominance of antropophilic and zoophilic species, respectively. These results are in agreement with statistical data from South and Southeast regions of Brazil, as well as from other parts of the world in which these fungi were the most frequently isolates from tinea pedis and tinea capitis. In this study, it was also observed a high percentage of T. tonsurans (41.7%) in tinea capitis and this result was different from the statistical data collected until now in our region.

Bioatividade de plantas medicinais no controle de Sclerotinia isolado de Stevia rebaudiana (Bert.) Bertoni

The microorganism Sclerotinia was isolated from roots of Stevia rebaudiana (Bert.) Bertoni in plantations in the northwest of Parana and submitted to the cultivation in the presence of extracts and vegetable balsams of Tarragon (Artemisia draconculus), Thyme (Thymus vulgaris), Manjerona (Origanum majorona), Mint citrata (Mintpiperita var. citrata), Purple Basil (Ocimum basilicum L.), Andiroba (Carapa guanensis) and Copaíba (Copaifera reticulata Ducke). The first five oils were extracted by steam drags, after the drying of the vegetable in greenhouse with circulation of air at 45°C. The last two were used in natura. A suspension (100ìl) of fungus previously cultivated, was added to each plate. The results show that after 7 days of incubation the thyme oils 10ìl, purple basil 25ìl, manjerona 25ìl, mint citrata 50ìl, tarragon 50ìl were capable to inhibit the growth of Sclerotinia, while the andiroba oil only reached this result with 200ìl. The copaiba balsam, even in the concentration of 500ìl, was unable to inhibit the growth of the microorganism.

Endophytic yeasts and filamentous fungi associated with southern Brazilian apple (Malus domestica) orchards subjected to conventional, integrated or organic cultivation

Endophytic fungi were isolated from leaves, flowers and fruit of healthy apple trees (Malus domestica, BORKH.) growing in southern Brazilian orchards under three different cultivation systems (conventional, integrated and organic), during two vegetative cycles. The greatest total number of endophytic isolates was obtained from the orchards under organic cultivation when compared to integrated and conventional cultivation systems. Filamentous fungi from the genera Colletotrichum, Xylaria and Botryosphaeria were the most frequent ones and the most representative yeast genera were Sporobolomyces, Rhodotorula, Debaryomyces and Cryptococcus. It is suggested that some isolates may be used as indicators of the different management systems. © 2005 Wiley-VCH Verlag GmbH & Co. KGaA.

Optimization of xylanase biosynthesis by Aspergillus japonicus isolated from a Caatinga area in the Brazilian state of Bahia

The objective of this research was to investigate the potential of xylanase production by Aspergillus japonicus and to determine the effects of cultivation conditions in the process, aiming toward optimization of enzyme production. The best temperature, as well as the best carbon source, for biomass production was determined through an automated turbidimetric method (Bioscreen-C). The enzyme activity of this fungus was separately evaluated in two solid substrates (wheat and soybean bran) and in Vogel medium, adding other carbon sources. Temperature effects, cultivation time, and spore concentrations were also tested. The best temperature for enzyme and biomass production was 25°C; however, the best carbon source for growth (determined by the Bioscreen C) did not turn out to be a good inducer of xylanase production. Maximum xylanase activity was achieved when the fungus was cultivated in wheat bran (without the addition of any other carbon source) using a spore concentration of 1 × 107 spores/mL (25°C, pH 5.0, 120 h). A. japonicus is a good xylanase producer under the conditions presented in these assays. © 2006 Academic Journals.

Fatty acid production by four strains of Mucor hiemalis grown in plant oil and soluble carbohydrates

Four Mucor hiemalis strains (M1, M2, M3 and M4), isolated from soil at a depth of 0 - 15 cm in the Juréia-Itatins Ecology Station (JIES), in the state of São Paulo, Brazil and were evaluated for the production of γ-linolenic (GLA) and other unsaturated fatty acids. Five growth variables (temperature, pH, carbon source, nitrogen source, and vegetable oils) were studied. Liquid media containing 2% vegetable oil (palm oil, canola oil, soybean oil, sesame oil, or sunflower oil) or 2% carbohydrate (fructose, galactose, glycerol, glucose, lactose, maltose, sucrose, sorbitol or xylose) and 1% yeast extract as a nitrogen source were used. The greatest biomass production was observed with M3 and M4 strains in palm oil (91.5 g l -1) and sunflower oil (68.3 g l -1) media, respectively. Strain M4 produced greater quantities of polyunsaturated acids in medium containing glucose. The GLA production in the M4 biomass was 1,132.2 mg l -1 in glucose medium. Plant oils were inhibitors of fatty acid production by these strains. © 2007 Academic Journals.

Production and partial characterization of keratinase produced by a microorganism isolated from poultry processing plant wastewater

A Streptomyces was isolated from poultry plant wastewater, showed high keratinolytic activity when cultured on feather meal medium. Optimum keratinolytic activity was observed at 40°C and pH 8.0. The enzyme also showed to be stable between 40 and 60°C. The keratinolytic activity was not inhibited by EDTA, DMSO and Tween 80. On the other hand, CaCl2, ZnCl2, and BaCl2 slightly inhibited the keratinolytic activity. The Streptomyces isolated might be useful in leather, keratin waste treatment, animal feeding industry, and also cosmetic industry. © 2008 Academic Journals.

Induction and secretion of elastinolytic and proteolytic activity in cultures of Paracoccidioides brasiliensis

P. brasiliensis parasitizes various human tissues and proteinases exported by this fungus may allow it to metabolize and invade host tissues. The influence of the culture medium on the production of proteinases by P. brasiliensis isolates was studied and the export of these enzymes was followed as a function of culture time. The fungus was grown in neopeptone, BSA, elastin or collagen medium. The culture medium was assayed for azocollytic, elastinolytic and caseinolytic activity. Proteolytic activity was also analysed by electrophoresis of the culture medium on gelatin and casein substrate gels. P. brasiliensis expressed relatively high levels of azocoll, elastin and casein degrading activity in all types of medium, except in neopeptone medium. Generally, expression of azocollytic activity peaked during the third week of culture and caseinolytic activity during the fourth week of culture. Azocollytic activity was highest at pH 4.0 and caseinolytic activity at pH 8.0. Elastinolytic activity was also highest at pH 8.0. This activity, as well as the others, may provide the fungus with a source of carbon and nitrogen and may also be responsible for the invasion of host tissues, such as pulmonary elastic fiber, by P. brasiliensis.