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.

Post-selection and return of foraged material by Acromyrmex subterraneus brunneus (Hymenoptera: Formicidae)

Two colonies of Acromyrmex subterraneus brunneus Forel (Hymenoptera: Formicidae) were studied regarding their behavior during cultivation of the fungus garden to determine a) the existence of post-selection of foraged material by the workers, and b) if present, the mechanism of this discrimination and how this material is returned. Many studies on plant processing by leaf-cutting ants have been carried out, but none of them has investigated the decision-making process of workers in the case of erroneous food selection. For this purpose, material with different degrees of moisture and hardness (floral sponge, polystyrene, plastic and clay) were individually offered to the colonies and the tasks performed by the different size categories were carefully recorded. Three tasks, i.e., foraging, cultivation of the fungus garden and return of the foraged material, were studied and subdivided into 14 subtasks. Analysis of all inert materials as a whole showed the presence of post-selection of foraged material through the return of material inadequate for the workers and the fungus. Discrimination of the inert material was observed at the time of shredding, probably based on parameters such as physical resistance to cutting and moisture content. A. s. brunneus workers showed flexibility in their activities during substrate processing. The observed post-selection of foraged material provides strong evidence for the cognitive abilities of worker ants and of the colony as a whole. Polymorphism and a complex society represent vital characteristics for the ecological success of this species.

The effect of trail length on task partitioning in three Acromyrmex species (Hymenoptera: Formicidae)

By comparing the behavior of three Acromyrmex (Hymenoptera, Formicidae) species during foraging on artificial trails of different lengths, we observed the occurrence of task partitioning and its relation to the food distance from the nest. Task partitioning was verified by leaf cache formation along the trail and leaf direct transferring among workers. There was significant difference between the number of leaf fragments carried directly to the fungus chamber and those transferred direct or indirectly, via cache, depending upon the trail length. Task partitioning could be a strategy used by leaf-cutting ants that allows the workers to use food sources far from their nests.

Effects of crude extracts of Agaricus blazei on DNA damage and on rat liver carcinogenesis induced by diethylnitrosamine

The effects of crude extracts of the mushroom Agaricus blazei Murrill (Agaricaceae) on both DNA damage and placental form glutathione S-transferase (GST-P)-positive liver foci induced by diethylnitrosamine (DEN) were investigated. Six groups of adult male Wistar rats were used. For two weeks, animals of groups 3 to 6 were treated with three aqueous solutions of A. blazei (mean dry weight of solids being 1.2, 5.6, 11.5 and 11.5 mg/ml, respectively). After this period, groups 2 to 5 were given a single ip injection 200 mg/kg DEN and groups 1 and 6 were treated with 0.9% NaCl. All animals were subjected to 70% partial hepatectomy at week five and sacrificed 4, 24 and 48 h or 8 weeks after DEN or 0.9% NaCl treatments (10th week after the beginning of the experiment). The alkaline comet assay and GST-P-positive liver foci development were used to evaluate the influence of the mushroom extracts on liver cell DNA damage and on the initiation of liver carcinogenesis, respectively. Previous treatment with the highest concentration of A. blazei (11.5 mg/ml) significantly reduced DNA damage, indicating a protective effect against DEN-induced liver cytotoxicity/genotoxicity. However, the same dose of mushroom extract significantly increased the number of GST-P-positive liver foci.

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.

Mango cultivars behavior in relation to fusarium subglutinans

The mango malformation, caused by the fungus Fusarium subglutinans Wollenweb & Reinking, is probably one of the diseases that causes more damages in the mangoes production in Brazil and other producing countries. This fungus was isolated of a Tommy Atkins plant with advanced symptoms of malformation, purified and prepared to be inoculated in 15 cultivars of national and imported mangoes. Initially, 10 cultivars had been inoculated in July/2000: Bourbon IAC - 100, Coração de Boi, Keitt, Parvin, Primor de Amoreira, Sensation, Smith, Surpresa, Tommy Atkins and Van Dyke. Another group was inoculated in December/2000, on the cultivars: Adams, Bhadauran, Palmer, Princesa and Zill, and with others cultivars Primor de Amoreira, Sensation and Tommy Atkins, that were inoculated in the first time, with the purpose to compare the two times of inoculation. Throughout the evaluated data at the two times of inoculation, after 11 months of evaluation, were maden the analysis of the variance in function of the involved sources of variation and the test of Duncan to compare the averages of inoculated cultivars. With the results obtained, the cultivars Bhadauran, Palmer, Parvin, Sensation, Van Dyke and Zill presents less percentage of plants with malformation symptoms or less progression of symptoms in relation to the others inoculated cultivars, under protected environmental conditions, where the assays were carried out.

Potential of Brazilian eugenia myrtaceae - As ornamental and as a fruit crop

Brazilian Myrtaceae comprises several genera of trees and shrubs used for ornamental and fruit production. In addition to the well known Guava, Pitanga and Jaboticaba, other species could be used for fruitculture, due the value and quality of their fruits and adaptation to some climate conditions mainly the subtropical one. Nine species of Eugenia were evaluated at Jaboticabal, located at 48° W and 21° S in São Paulo state in a germplasm bank. The average rain by year is 1431 mm and the temperature 22,2° C at an altitude of 575 m. The species are Eugenia klozschiana Berg. (Pero-do-campo), E. stipitata Mc Vaugh (Araça-boi), E. tomentosa Camb. (Cabeludinha), E. dysentherica DC. (Cagaita), E. brasiliensis Berg. (Grumixama), E.pitanga (Pitanga-anã), E. luchsnathiana Berg. (Pitomba), E. uvalha Camb. (Uvaia) and E. involucrata DC. (Cereja-do-rio-grande). The evaluations comprised tree development, fruit quality and leaf and flower morphological studies. The main results are: the trees of Pera-do-campo and Pitanga-anã are small shrubs of 1 to 2 m height, Araça-boi and Cabeludinha are small trees, 3 to 5 m high, and the other species are tall trees, with 5 to 10 m height. The species adapted well to the subtropical conditions, except for Araça-boi, which is native to the Amazonian region and exhibited a severe fungus disease infection. In relation to fruit quality, all the species had edible fruit, some were sweet and juicy, Cabeludinha, Grumixama, Pitomba, Cereja-do-rio grande and Pitanga-anã, while others had high acidity (Araça-boi, Pera-do-campo, Cagaita and Uvaia and were more suitable for processing. Simple, single leaves were characteristic of all species, but with different sizes and shapes., With the addition of color, smell and other characteristics, leaf size and shape were useful for comparative classification. Flower components and structure are described also.

rDNA-RFLP identification of Candida species in immunocompromised and seriously diseased patients

PCR was used to amplify a targeted region of the ribosomal DNA of 76 Candida spp. isolates from immunocompromised and seriously diseased patients. Thirty-seven strains isolated from different anatomical sites of 11 patients infected with HIV (Vitória, ES, Brazil), 26 isolates from patients under treatment at Odilon Behrens Hospital and 13 isolates from skin and urine samples from São Marcos Clinical Analysis Laboratory (Belo Horizonte, Brazil) were scored. Fragments of rDNA were amplified using primer pairs ITS1-ITS4, for the amplification of ITS1 and ITS2 regions, including the gene for the 5.8s subunit. Amplification resulted in fragments ranging in size from 350 to 950 bp. Amplicons were digested with eight restriction enzymes. A pattern of species-specificity among the different medically important Candida species could be identified following restriction digestion of the PCR products. Candida albicans was the species most frequently observed, except for the group of newborns under treatment at the Odilon Behrens Hospital and for the isolates from the clinical analysis laboratory. C. parapsilosis was the species most frequently observed in these two groups.

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.

Avaliação da formação do biofilme de Candida albicans sobre superfície de titânio revestida com hidroxiapatita por meio de microscopia eletrônica de varredura

In this study Candida albicans biofilm formation on the surface of commercially pure titanium (cp-Ti) coated with hydroxyapatite was observed by means of scanning electron microscope. The biofilm was formed after 45 days of incubation of the samples in liquid culture medium inoculated with fungus cells in a tube of polystyrene with screw cap and sterilized. After the biofilm removal with 10% EDTA solution was observed pitting on the surface of cp-Ti coated.

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.

Purification and biochemical characterization of two xylanases produced by Aspergillus caespitosus and their potential for kraft pulp bleaching

Two extracellular xylanases produced by the thermotolerant fungus Aspergillus caespitosus grown in sugar cane bagasse were purified and characterized. Estimated molecular masses were 26.3 and 27 kDa (xyl I); 7.7 and 17.7 kDa (xyl II) for gel filtration and SDS-PAGE, respectively. Optimal temperature for both xylanases was 50-55°C. Optimal pH was 6.5-7.0 for xyl I, and 5.5-6.5 for xyl II. The thermostability (T half) at 55°C was 27.3 min (xyl I) and >90 min (xyl II). Xylanase activity was inhibited by several ions. β-mercaptoethanol activated 59 and 102% xyl I and xyl II activities, respectively. These enzymes preferentially hydrolyzed birchwood xylan, and the K m and V max values were 2.5 mg/ml and 1679 U/mg protein (xyl I), and 3.9 mg/ml and 113 U/mg protein (xyl II). The action of both xylanases mainly that of xyl II, on kraft pulp reduced kappa number and increased pulp viscosity. © 2004 Elsevier Ltd. All rights reserved.