Spathaspora marinasilvae sp. nov., a xylose-fermenting yeast isolated from galleries of passalid beetles and rotting wood in the Amazonian rainforest biome.

Four yeast isolates were obtained from rotting wood and galleries of passalid beetles collected in different sites of the Brazilian Amazonian Rainforest in Brazil. This yeast produces unconjugated allantoid asci each with a single elongated ascospore with curved ends. Sequence analysis of the internal transcribed spacer-5.8 S region and the D1/D2 domains of the large subunit ribosomal RNA (rRNA) gene showed that the isolates represent a novel species of the genus Spathaspora. The novel species is phylogenetically related to a subclade containing Spathaspora arborariae and Spathaspora suhii. Phylogenomic analysis based on 1884 single-copy orthologs for a set of Spathaspora species whose whole genome sequences are available confirmed that the novel species represented by strain UFMG-CM-Y285 is phylogenetically close to Sp. arborariae. The name Spathaspora marinasilvae sp. nov. is proposed to accommodate the novel species. The holotype of Sp. marinasilvae is CBS 13467 T (MycoBank 852799). The novel species was able to accumulate xylitol and produce ethanol from d-xylose, a trait of biotechnological interest common to several species of the genus Spathaspora.

Yueomyces silvicola sp. nov., a novel ascomycetous yeast species unable to utilize ammonium, glutamate, and glutamine as sole nitrogen sources.

Five yeast strains isolated from tree bark and rotten wood collected in central and southwestern China, together with four Brazilian strains (three from soil and rotting wood collected in an Amazonian rainforest biome and one from Bromeliad collected in Alagoas state) and one Costa Rican strain isolated from a flower beetle, represent a new species closely related with Yueomyces sinensis in Saccharomycetaceae, as revealed by the 26S ribosomal RNA gene D1/D2 domain and the internal transcribed spacer region sequence analysis. The name Yueomyces silvicola sp. nov. is proposed for this new species with the holotype China General Microbiological Culture Collection Center 2.6469 (= Japan Collection of Microorganisms 34885). The new species exhibits a whole-genome average nucleotide identity value of 77.8% with Y. sinensis. The two Yueomyces species shared unique physiological characteristics of being unable to utilize ammonium and the majority of the amino acids, including glutamate and glutamine, as sole nitrogen sources. Among the 20 amino acids tested, only leucine and tyrosine can be utilized by the Yueomyces species. Genome sequence comparison showed that GAT1, which encodes a GATA family protein participating in transcriptional activation of nitrogen-catabolic genes in Saccharomyces cerevisiae, is absent in the Yueomyces species. However, the failure of the Yueomyces species to utilize ammonium, glutamate, and glutamine, which are generally preferred nitrogen sources for microorganisms, implies that more complicated alterations in the central nitrogen metabolism pathway might occur in the genus Yueomyces.

Saccharomycopsis praedatoria sp. nov., a predacious yeast isolated from soil and rotten wood in an Amazonian rainforest biome.

Three yeast isolates were obtained from soil and rotting wood samples collected in an Amazonian rainforest biome in Brazil. Comparison of the intergenic spacer 5.8S region and the D1/D2 domains of the large subunit rRNA gene showed that the isolates represent a novel species of the genus Saccharomycopsis. A tree inferred from the D1/D2 sequences placed the novel species near a subclade containing Saccharomycopsis lassenensis, Saccharomycopsis fermentans, Saccharomycopsis javanensis, Saccharomycopsis babjevae, Saccharomycopsis schoenii and Saccharomycopsis oosterbeekiorum, but with low bootstrap support. In terms of sequence divergence, the novel species had the highest identity in the D1/D2 domains with Saccharomycopsis capsularis, from which it differed by 36 substitutions. In contrast, a phylogenomic analysis based on 1061 single-copy orthologs for a smaller set of Saccharomycopsis species whose whole genome sequences are available indicated that the novel species represented by strain UFMG-CM-Y6991 is phylogenetically closer to Saccharomycopsis fodiens and Saccharomycopsis sp. TF2021a (=Saccharomycopsis phalluae). The novel yeast is homothallic and produces asci with one spheroidal ascospore with an equatorial or subequatorial ledge. The name Saccharomycopsis praedatoria sp. nov. is proposed to accommodate the novel species. The holotype of Saccharomycopsis praedatoria is CBS 16589T. The MycoBank number is MB849369. S. praedatoria was able to kill cells of Saccharomyces cerevisiae by means of penetration with infection pegs, a trait common to most species of Saccharomycopsis.

Sugiyamaella bielyi f. a., sp. nov. and Sugiyamaella amazoniana f. a., sp. nov., two yeast species isolated from passalid beetles and rotting wood in Amazonia.

Sixteen yeast isolates representing two novel species of the genus Sugiyamaella were obtained from passalid beetles, their galleries and rotting wood collected in three sites of Amazonian Forest in Brazil. Sequence analyses of the ITS-5.8S region and the D1/D2 domains of the large subunit rRNA gene showed that the first species, described here as Sugiyamaella amazoniana f. a., sp. nov. (holotype CBS 18112, MycoBank 847461) is phylogenetically related to S. bonitensis with these species differing by 37 nucleotide substitutions and six gaps in D1/D2 sequences. S. amazoniana is represented by nine isolates obtained from the guts of the passalid beetles Popilius marginatus, Veturius magdalenae, Veturius sinuosus and Spasalus aquinoi, a beetle gallery and rotting wood. The second species, Sugiyamaella bielyi f. a., sp. nov. (holotype CBS 18148, MycoBank 847463), is most phylogenetically related to several undescribed Sugiyamaella species. S. bielyi is described based on seven isolates obtained from the guts of V. magdalenae and V. sinuosus, a beetle gallery and rotting wood. Both species appear to be associated with passalid beetles and their ecological niches in Amazonian biome.

The Brazilian Amazonian rainforest harbors a high diversity of yeasts associated with rotting wood, including many candidates for new yeast species.

This study investigated the diversity of yeast species associated with rotting wood in Brazilian Amazonian rainforests. A total of 569 yeast strains were isolated from rotting wood samples collected in three Amazonian areas (Universidade Federal do Amazonas-Universidade Federal do Amazonas [UFAM], Piquiá, and Carú) in the municipality of Itacoatiara, Amazon state. The samples were cultured in yeast nitrogen base (YNB)-d-xylose, YNB-xylan, and sugarcane bagasse and corncob hemicellulosic hydrolysates (undiluted and diluted 1:2 and 1:5). Sugiyamaella was the most prevalent genus identified in this work, followed by Kazachstania. The most frequently isolated yeast species were Schwanniomyces polymorphus, Scheffersomyces amazonensis, and Wickerhamomyces sp., respectively. The alpha diversity analyses showed that the dryland forest of UFAM was the most diverse area, while the floodplain forest of Carú was the least. Additionally, the difference in diversity between UFAM and Carú was the highest among the comparisons. Thirty candidates for new yeast species were obtained, representing 36% of the species identified and totaling 101 isolates. Among them were species belonging to the clades Spathaspora, Scheffersomyces, and Sugiyamaella, which are recognized as genera with natural xylose-fermenting yeasts that are often studied for biotechnological and ecological purposes. The results of this work showed that rotting wood collected from the Amazonian rainforest is a tremendous source of diverse yeasts, including candidates for new species.

8-Hydroxyquinoline 1,2,3-triazole derivatives with promising and selective antifungal activity.

Fungal infections that affect humans and plants have increased significantly in recent decades. However, these pathogens are still neglected when compared to other infectious agents. Due to the high prevalence of these infections, the need for new molecules with antifungal potential is recognized, as pathogenic species are developing resistance to the main drugs available. This work reports the design and synthesis of 1,2,3-triazole derivatives of 8-hydroxyquinoline, as well as the determination of their activities against a panel of fungal species: Candida spp., Trichosporon asahii, Magnusiomyces capitatus, Microsporum spp., Trichophyton spp. and Fusarium spp. The triazoles 5-(4-phenyl-1H-1,2,3-triazol-1-yl)quinolin-8-ol (12) and 5-(4-(cyclohex-1-en-1-yl)-1H-1,2,3-triazol-1-yl)quinolin-8-ol (16) were more promising, presenting minimum inhibitory concentration (MIC) values between 1-16 µg/ml for yeast and 2-4 µg/ml for dermatophytes. However, no relevant anti-Fusarium spp. activity was observed. In the time-kill assays with Microsporum canis, 12 and 16 presented time-dependent fungicide profile at 96 h and 120 h in all evaluated concentrations, respectively. For Candida guilliermondii, 12 was fungicidal at all concentrations at 6 h and 16 exhibited a predominantly fungistatic profile. Both 12 and 16 presented low leukocyte toxicity at 4 µg/ml and the cell viability was close to 100% after the treatment with 12 at all tested concentrations. The sorbitol assay combined with SEM suggest that damages on the fungal cell wall could be involved in the activity of these derivatives. Given the good results obtained with this series, scaffold 4-(cycloalkenyl or phenyl)-5-triazol-8-hydroxyquinoline appears to be a potential pharmacophore for exploration in the development of new antifungal agents.

Anti-Candida and anti-quorum sensing activity of airborne microorganisms detected by a rapid method.

INTRODUCTION: Introducing new antibiotics to the clinic is critical. METHODS: We adapted a plate method described by Kawaguchi and coworkers in 20131 for detecting inhibitory airborne microorganisms. RESULTS: We obtained 51 microbial colonies antagonist to Chromobacterium violaceum, purified and retested them, and of these, 39 (76.5%) were confirmed. They comprised 24 bacteria, 13 fungi, and 2 yeasts. Among the fungi, eight (61.5%) produced active extracts. Among the bacterial, yeast, and fungal strains, 17 (44.7%) and 12 (31.6%) were active against Candida albicans and Candida parapsilosis, respectively. CONCLUSIONS: The proposed screening method is a rapid strategy for discovering potential antibiotic producers.

Anti-Candida and anti-quorum sensing activity of airborne microorganisms detected by a rapid method

Abstract INTRODUCTION: Introducing new antibiotics to the clinic is critical. METHODS: We adapted a plate method described by Kawaguchi and coworkers in 20131 for detecting inhibitory airborne microorganisms. RESULTS: We obtained 51 microbial colonies antagonist to Chromobacterium violaceum, purified and retested them, and of these, 39 (76.5%) were confirmed. They comprised 24 bacteria, 13 fungi, and 2 yeasts. Among the fungi, eight (61.5%) produced active extracts. Among the bacterial, yeast, and fungal strains, 17 (44.7%) and 12 (31.6%) were active against Candida albicans and Candida parapsilosis, respectively. CONCLUSIONS: The proposed screening method is a rapid strategy for discovering potential antibiotic producers.

Microbiological and chemical characteristics of tarubá, an indigenous beverage produced from solid cassava fermentation.

The aim of this work was to identify and characterize the microbiota present during fermentation and in the final beverage, tarubá, by culture-dependent and -independent methods. In addition, target chemical compounds (carbohydrates, organic acids, and ethanol) were evaluated. Lactic acid bacteria (LAB) and mesophilic bacteria were the predominant microorganisms. Among them, Lactobacillus plantarum, Lactobacillus brevis, Leuconostoc mesenteroides, and Bacillus subtilis were frequently isolated and detected by DGGE analysis. Torulaspora delbrueckii was the dominant yeast species. Yeast isolates Pichia exigua, Candida rugosa, T. delbrueckii, Candida tropicalis, Pichia kudriavzevii, Wickerhamomyces anomalus, and Candida ethanolica and bacteria isolates Lb. plantarum, Bacillus subtilis, Bacillus amyloliquefaciens, Bacillus licheniformis, Bacillus sp., and Chitinophaga terrae showed amylolytic activity. Only isolates of P. exigua and T. delbrueckii and all species of the genus Bacillus identified in this work exhibited proteolytic activity. All microbial isolates grew at 38 °C, and only the isolates belonging to Hanseniaspora uvarum species did not grow at 42 °C. These characteristics are important for further development of starter cultures; isolates of T. delbrueckii, P. exigua, and Bacillus species identified in this work displayed all of these properties and are potential strains for use as starter culture in cassava fermented food.

Glutathione levels in and total antioxidant capacity of Candida sp. cells exposed to oxidative stress caused by hydrogen peroxide.

INTRODUCTION: The capacity to overcome the oxidative stress imposed by phagocytes seems to be critical for Candida species to cause invasive candidiasis. METHODS: To better characterize the oxidative stress response (OSR) of 8 clinically relevant Candida sp., glutathione, a vital component of the intracellular redox balance, was measured using the 5,5'-dithiobis-(2-nitrobenzoic acid (DTNB)-glutathione disulfide (GSSG) reductase reconversion method; the total antioxidant capacity (TAC) was measured using a modified method based on the decolorization of the 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic) acid radical cation (ABTS*+). Both methods were used with cellular Candida sp. extracts treated or not with hydrogen peroxide (0.5 mM). RESULTS: Oxidative stress induced by hydrogen peroxide clearly reduced intracellular glutathione levels. This depletion was stronger in Candida albicans and the levels of glutathione in untreated cells were also higher in this species. The TAC demonstrated intra-specific variation. CONCLUSIONS: Glutathione levels did not correlate with the measured TAC values, despite this being the most important non-enzymatic intracellular antioxidant molecule. The results indicate that the isolated measurement of TAC does not give a clear picture of the ability of a given Candida sp. to respond to oxidative stress.

Glutathione levels in and total antioxidant capacity of Candida sp. cells exposed to oxidative stress caused by hydrogen peroxide / Níveis de glutationa e capacidade antioxidante total em células de Candida sp. expostas a estresse oxidativo causado por peróxido de hidrogênio

INTRODUCTION: The capacity to overcome the oxidative stress imposed by phagocytes seems to be critical for Candida species to cause invasive candidiasis. METHODS: To better characterize the oxidative stress response (OSR) of 8 clinically relevant Candida sp., glutathione, a vital component of the intracellular redox balance, was measured using the 5,5'-dithiobis-(2-nitrobenzoic acid (DTNB)-glutathione disulfide (GSSG) reductase reconversion method; the total antioxidant capacity (TAC) was measured using a modified method based on the decolorization of the 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic) acid radical cation (ABTS*+). Both methods were used with cellular Candida sp. extracts treated or not with hydrogen peroxide (0.5 mM). RESULTS: Oxidative stress induced by hydrogen peroxide clearly reduced intracellular glutathione levels. This depletion was stronger in Candida albicans and the levels of glutathione in untreated cells were also higher in this species. The TAC demonstrated intra-specific variation. CONCLUSIONS: Glutathione levels did not correlate with the measured TAC values, despite this being the most important non-enzymatic intracellular antioxidant molecule. The results indicate that the isolated measurement of TAC does not give a clear picture of the ability of a given Candida sp. to respond to oxidative stress.

INTRODUÇÃO: A capacidade de suportar o estresse oxidativo imposto por fagócitos parece ser crítica para que espécies de Candida causem candidíase invasiva. MÉTODOS: Para melhor caracterizar a resposta ao estresse oxidativo (REO) de oito Candida sp. clinicamente relevantes, um componente vital do balanço redox intracelular, a glutationa, foi mensurada pelo método de reconversão DTNB-GSSG redutase e a capacidade antioxidante total (CAT) foi mensurada por um método modificado baseado na descoloração do ABTS*+. Ambos os métodos foram utilizados em extratos celulares das espécies de Candida tratadas ou não com peróxido de hidrogênio (0,5mM). RESULTADOS: O estresse oxidativo induzido pelo peróxido de hidrogênio claramente reduziu os níveis intracelulares de glutationa. Esta diminuição foi mais intensa em C. albicans e os níveis de glutationa em células não tratadas foram também maiores nesta espécie. A capacidade antioxidante total demonstrou variação intraespecífica na capacidade antioxidante. CONCLUSÕES: Os níveis de glutationa não se correlacionaram com a capacidade antioxidante total mensurada, apesar desta ser a defesa antioxidante intracelular não-enzimática mais importante. Os resultados indicam que a medição isolada da CAT não fornece um quadro claro da habilidade de certa espécie de Candida responder ao estresse oxidativo.

Differential resistance to oxidants and production of hydrolytic enzymes in Candida albicans.

Resistance to the toxic effects of reactive oxygen species produced by phagocytes and production of hydrolytic enzymes are important aspects of Candida albicans virulence. In this report, we compared twelve C. albicans isolates for their in vitro capacity to resist oxidants-hydrogen peroxide, menadione and paraquat; and to produce hydrolytic enzymes-phospholipase and protease. Different C. albicans isolates showed different degrees of resistance to oxidants as well as differences in production of hydrolytic enzymes. Resistance to oxidative stress did not correlate with production of hydrolytic enzymes. This reinforces the view that C. albicans differentially regulates the expression of virulence factors in response to local environmental conditions.

Avaliação de infecção urinária em gestantes do município de Marechal Cândido Rondon - PR / Evaluation of urinary infection in pregnant women in the municipality of marechal cândido rondon - PR

A infecção urinária constitui uma das intercorrências clínicas mais comuns nas gestantes, devido a alterações anatômicas e fisiológicas que ocorrem neste período, facilitando o seu desenvolvimento. Durante a gravidez, infecções urinárias mais graves podem causar contrações do útero, levando ao trabalho de parto prematuro, aborto, hipertensão arterial, morte do bebê e até mesmo da mãe quando a infecção se torna severa e generalizada. Desse modo, o objetivo desse trabalho foi avaliar a incidência de infecção urinária, verificando em que período gestacional está mais presente, além de identificar os principais microrganismos envolvidos e avaliar a sintomatologia nas infecções positivas. A pesquisa foi realizada por meio do exame de urocultura de 34 gestantes. Das amostras analisadas, 56% apresentaram resultado positivo, e dentre as positivas 63,1% tiveram o desenvolvimento de Escherichia coli. As gestantes que estavam no 3° trimestre gestacional apresentaram maior incidência de infecção urinária, constatada em 71,4% das pesquisadas. Apenas 16% das gestantes com infecção urinária, apresentaram sinais e sintomas que indicassem a suspeita de infecção. A presença de infecção urinária assintomática na maioria das gestantes demonstra a importância da realização do exame de urocultura no período gestacional, a fim de diagnosticar e tratar precocemente as infecções urinárias, evitando possíveis complicações.

The urinary infection is one of the most common clinical complications in pregnant women due to anatomic and physiological changes that occur in this period, facilitating their development. During pregnancy, urinary infections can cause more severe contractions of the uterus, leading to a work of premature birth, abortion, hypertension, death of the baby and even of the mother when the infection becomes severe and widespread. The aim of this study is to verify the incidence of urinary tract infection, verify that pregnancy is present, it is identify the key microorganisms and evaluate the positive symptoms of infections. The survey was conducted by examining urine of 34 pregnant women. Of the samples analyzed, 56% were positive, and among the 63.1% had positive development of Escherichia coli. The women in the 3 rd trimester of pregnancy showed a higher incidence of urinary tract infection, found in 71.4% of the surveyed women. Only 16% of pregnant women with urinary infection, had signs and symptoms that indicate a suspicion of urinary tract infection. The presence of asymptomatic urinary infection in most pregnant women demonstrates the importance of completing the examination of urine culture in the gestational period in order to diagnose and treat urinary infections early, avoiding possible complications.

Response to oxidative stress in eight pathogenic yeast species of the genus Candida.

In the course of an infection, the formation of reactive oxygen species by phagocytes and the antioxidant defense mechanisms of microorganisms play a crucial role in pathogenesis. In this study, isolates representing 8 pathogenic Candida species-Candida albicans, Candida dubliniensis, Candida famata, Candida glabrata, Candida guilliermondii, Candida krusei, Candida parapsilosis and Candida tropicalis-were compared with regard to their resistance to oxidative stress in vitro. We evaluated degree of resistance, induction of oxidative damage, capacity to adapt, and induction of antioxidant enzymes. The species showed variable sensitivity to oxidative attack. C. albicans, C. glabrata, and C. krusei were more resistant to oxidative stress under the conditions tested; C. parapsilosis and C. tropicalis presented medium resistance; and C. dubliniensis, C. famata, and C. guilliermondii were more sensitive. The overall greater resistance to oxidative stress of C. albicans and C. glabrata may provide an advantage to these species, which are the major causative agents of candidiasis.

[Production of virulence factors in vitro by pathogenic species of the genus Candida]. / Produção de fatores de virulência in vitro por espécies patogênicas do gênero Candida.

The growth capacity at 39 degrees C and 42 degrees C, production of hydrolytic enzymes and hemolytic activity of 21 clinical and reference strains of seven species of Candida spp were evaluated in vitro.Candida dubliniensis and Candida krusei demonstrated lower virulence potential and Candida albicans higher potential.

Produção de fatores de virulência in vitro por espécies patogênicas do gênero Candida / Production of virulence factors in vitro by pathogenic species of the genus Candida

Avaliou-se, in vitro, a capacidade de crescimento em 39ºC e 42ºC, a produção de enzimas hidrolíticas e a atividade hemolítica de 21 cepas clínicas e de referência de sete espécies de Candida spp, Candida dubliniensis e Candida krusei demonstraram menor potencial de virulência e Candida albicans maior.

The growth capacity at 39ºC and 42ºC, production of hydrolytic enzymes and hemolytic activity of 21 clinical and reference strains of seven species of Candida spp were evaluated in vitro.Candida dubliniensis and Candida krusei demonstrated lower virulence potential and Candida albicans higher potential.

Avaliação da qualidade microbiológica das drogas vegetais mais comercializadas em farmácias demanipulação de Toledo-PR / Evaluation of the microbiological quality of the most commercialized herbal drugs in

A qualidade microbiológica de produtos vegetais constitui atributo essencial para seu desempenho adequado. Neste estudo, foram avaliadas 27 amostras de três espécies vegetais: ginko biloba (Ginkgo biloba), cáscara sagrada (Rhamnus purshiana) e sene (Cassiaangustifolia). O método utilizado foi o de semeadura em profundidade (pour plate) e foram realizadas contagens em placas de Petri com meios seletivos e pesquisa de microrganismos específicos, de acordo com a Organização Mundial da Saúde (OMS). A contaminação bacterianavariou de 0,5 x 101 a 9,0 x 104 unidades formadoras de colônia (UFC)/g, estando todas as amostras em conformidade com a OMS e 5 (18,5 por cento), acima dos limites permitidos pela Farmacopéia Brasileira (FB). A contaminação fúngica variou de 0,5 x 101 a 8,45 x 104 UFC/g, com 6 (22,2 por cento) amostras acima dos limites da OMS e 9 (33,3 por cento) acima dos limites da FB. Foi confirmada a presença de Escherichia coli em 2 (7,4 por cento) amostras e Staphylococcus aureus em 6 (22,2 por cento). Considerando as recomendações da OMS, 22,2 por cento das matérias-primas analisadas foram consideradas insatisfatórias para o consumo e, de acordo com a FB, 37 das amostras apresentaram populações bacterianase fúngicas acima do tolerável, ou continham bactérias não aceitas. Os resultados apontam para a necessidade de controle microbiológico mais rigoroso das drogas vegetais.

The microbiological quality of vegetal products constitutes one of the essential attributes for its adequate performance. In this study, 27 vegetal raw material samples of three species, Ginkgo biloba, Rhamnus purshiana and Cassia angustifolia, were evaluated.The method utilized was pour plate plating and colony counts in Petri dishes with selective medium and investigation of specific microorganism were carried out in accordance with the World Health Organization (WHO). Bacterial contamination varied from 0.5 x 101 to 9.0 x 104 colony-forming units (CFU)/g, all samples in accordance with WHO and 5 (18.5 percent), above the limits allowed by the Brazilian Pharmacopoeia(FB). Fungal contamination varied from 0.5 x 101 to 8.45 x 104 CFU/g, with 6 (22.2 percent) samples above of the limits of WHO and 9 (33.3 percent) above of the limits of the FB. Presence of Escherichia coli in 2 (7.4 percent) and Staphylococcus aureus in 6 (22.2 percent) was confirmed.Considering the recommendations of WHO, 22.2 percent of the analyzed raw materials were considered unsatisfactory for consumption, and, inaccordance with the FB, 37 percent of the samples presented bacterial and fungal populations either above the tolerated or contained unaccepted bacteria. The results point out to the need for more rigorous microbiological control of herbal drugs.

Avaliação da utilização de antibióticos por profissionais de saúde e pela população na cidade de toledo, Paraná, Brasil / Assessment of the use of antibiotics by health professionals and the population in toledo - R - Brazil

Os princípios básicos do uso de antibióticos estão bem estabelecidos. Contudo, existem relatos de uso inadequado destas drogas, o que parece estar relacionado com o desenvolvimento de cepas resistentes de microrganismos patogênicos. Considerando o exposto, neste estudo foi avaliado o emprego de antibióticos por profissionais de saúde, bem como a utilização destes fármacos pela população do município de Toledo, Paraná, através de questionários com respostas objetivas. Dentre as informações obtidas, ressalta-se que: a escolha da droga empregada pelos médicos é baseada, em regra, somente no exame clínico; 79,4% dos médicos e 87,5% dos analistas clínicos entrevistados relataram ter observado aumento nos eventos de resistência a antibióticos; 57,6% dos farmacêuticos entrevistados em farmácias de dispensação afirmaram que antibióticos são dispensados sem prescrição médica; 81,4% destes farmacêuticos relataram que a população solicita antibióticos para doenças de etiologia viral, como gripes e resfriados; 52,3% dos membros da população entrevistada dizem ter recebido indicação de antibióticos na farmácia e 57,5% de parentes, amigos e vizinhos; somente 47% dos entrevistados referiram realizar a antibioticoterapia completa; todos os médicos e 74,6% dos farmacêuticos em farmácias acreditam que a população não utiliza antibióticos de forma correta. Os dados obtidos evidenciam má utilização deste grupo de medicamentos. Faz-se necessário informar a população e os profissionais da saúde sobre a importância da utilização discriminada de antibióticos para evitar reflexos nos custos em saúde.

The basic principles of the use of antibiotics are well established, although there are reports of inadequate utilization of these drugs, which seems to be related with the development of resistant strains of pathogenic microorganisms. The employment of antibiotics by Health Professionals and the utilization of these drugs by the population in Toledo-PR were assessed through questionnaires. It standsout among the information obtained that: the choice of the drug by the Physicians is normally based only on the clinic examination; 79.4% of the Physicians interviewed, and 87.5% of the Clinical Analysts mentioned to have observed an increase in the events of resistance to antibiotics; 57.6% of the Pharmacists interviewed in Drugstores mentioned that antibiotics are dispensed without prescription; 81.4% of these Pharmacists mentioned that the population asks for antibiotics for diseases of viral etiologies such as the fl u and ordinary cold; 52.3% of the population interviewed said that they had received indication of antibiotics in the Drugstore, and 57.5% by relatives, friends and neighbors; just 47% of the interviewed in the population mentioned to have carried out the complete treatment; all the Physicians and 74.6%of the Pharmacists in Drugstores believed that the population do not use antibiotics correctly. The data obtained shows bad utilization of this group of medicines. It is necessary to inform both population and Health Professionals about the importance of the indiscriminate use of antibiotics to avoid refl ex on the health costs.

Cryptococcus neoformans and Cryptococcus gattii isolated from the excreta of psittaciformes in a southern Brazilian zoological garden.

Cryptococcus neoformans, a major pathogen in immunocompromised patients, is a ubiquitous free-living fungus that can be isolated from soils, avian excreta and plant material. To further study potential saprophytic sources of this yeast in the Southern Brazilian State Rio Grande do Sul, we analyzed fecal samples from 59 species of captive birds kept in cages at a local Zoological Garden, belonging to 12 different orders. Thirty-eight environmental isolates of C. neoformans were obtained only from Psittaciformes (Psittacidae, Cacatuidae and Psittacula). Their variety and serotype were determined, and the genetic structure of the isolates was analyzed by use of the simple repetitive microsatellite specific primer M13 and the minisatellite specific primer (GACA)(4) as single primers in the PCR. The varieties were confirmed by pulsed-field gel electrophoresis (PFGE). Thirty-three isolates (87%) were from the var. grubii, serotype A, molecular type VNI and five (13%) were Cryptococcus gattii, serotype B, molecular type VGI. All the isolates were mating type alpha. Isolates were screened for some potential virulence factors. Quantitative urease production by the environmental isolates belonging to the C. gattii was similar to the values usually obtained for clinical ones.