Susceptibilidad de levaduras vaginales a los antifúngicos más utilizados en Maringá, Paraná, Brasil / Susceptibility to vaginal yeast in most used antifungal in Maringá, Paraná, Brazil

Se determinó la susceptibilidad antifúngica in vitro de 78 cepas de levaduras aisladas de mujeres de la ciudad de Maringá/Paraná, Brasil, con candidiasis vulvovaginal (CVV), atendidas en el Laboratorio de Enseñanza e Investigación en Análisis Clínicos (LEPAC) de la Universidad Estatal de Maringá, desde el 1 de enero 2005 al 31 de diciembre 2006. Su sensibilidad in vitro fue investigada por el método de microdilución frente a ketoconazol (KETO), fluconazol (FLU), itraconazol (ITRA), nistatina (NIS) y anfotericina B (AMB). Para KETO, 41,5% de las cepas de C. albicans y 96% de Candida no-albicans presentaron resistencia (100% de C. glabrata) y para FLU solamente el 3,8% de los aislamientos de C. albicans y el 8,0% de C. glabrata fueron resistentes. Sólo 1,9% de las cepas de C. albicans y 20% de las de C. no-albicans fueron resistentes a ITRA y el 5,7% de las C. albicans y el 8% de las C. no-albicans (sólo C. glabrata) fueron resistentes a AMB. No hubo aislamientos resistentes a NIST, pero sí una elevada frecuencia de sensibilidad dosis dependiente "in vitro". Estos datos avalan la creciente necesidad de la realización de pruebas de identificación y susceptibilidad in vitro a los antifúngicos para establecer el correcto tratamiento de la CVV.

In vitro antifungal susceptibility was determined in the 78 yeasts isolated from patients with vulvovaginal candidiasis (VVC) from the city of Maringá/Parana/Brazil, assisted in the Laboratory of Teaching and Research in Clinical Analysis of the State University of Maringá, from 01 January 2005 to December 31, 2006. Its sensibility in vitro was tested according to microdilution method in front of ketoconazol (KETO), fluconazole (FLU), itraconazole (ITRA), nistatin (NIS) and amphotericin B (AMB). For KET, 41.5% of the C. albicans and 96.0% of the C. non-albicans showed resistance (100.0% of C. glabrata) and for FLU, only 3.8% of the isolates of C. albicans and 8.0% of C. glabrata showed resistance. Only 1.9% of the C. albicans and 20% of the C. no-albicans were resistant. For AMB, 5.7% of the C. albicans and 8% of the C. no-albicans (only C. glabrata), were resistant. There were no isolations resistant from NIST, however, there was a high frequency of dose-dependent sensibility (SDD) in vitro. These data makes it possible to confirm the growing necessity of the performance of identification tests and in vitro antifungal susceptibility to antifungals to establish the correct treatment of CVV.

Lack of association between genotypes and virulence factors in C. albicans strains isolated from vaginal secretion

The physiopathogenesis of vulvovaginal candidiasis (VVC) is still not completely elucidated. The objective of this study was to evaluate if there is a relationship between the different genotypes of Candida albicans, their main agent and the virulence of this yeast in vaginal isolates, and to check if there are laboratorial markers that can predict the ability of each isolate to develop VVC independently of symptoms. The production of exoenzymes protease, phospholipase and haemolysin, resistance to hydrogen peroxide, and the genotype were determined. Genotype A was predominant (75 percent), protease, phospholipase and haemolytic activity were highly expressed, and the majority of the yeasts were sensitive to H2O2 in 1 and 2 hours of exposure, suggesting that these factors are important in the virulence of vaginal isolates. However they did not have any correlation with the genotypes. The different isolates expressed similar virulence potential, suggesting that other factors relating to the yeasts and the host must participate in the development of the clinical disease.

A fisiopatogenia da candidíase vulvovaginal (CVV) não está completamente elucidada até o presente momento. O objetivo deste estudo foi avaliar se exite relação entre os diferentes genótipos de Candida albicans, seu principal agente, e a virulência desta levedura em isolados vaginais, e checar se existem marcadores laboratoriais que possam predizer a habilidade de cada isolado para desenvolver CVV independentemente dos sintomas. Foram determinados a produção de exoenzimas protease, fosfolipase and hemolisina, resistência ao peróxido de hidrogêncio, e genótipo. O genótipo A foi predominante (75 por cento), protease, fosfolipase e atividade hemolítica foram alevadamente expressos, e a maioria das leveduras foram sensíveis ao H2O2 em 1 e 2 horas de exposição, sugerindo que estes fatores são importantes na virulênciae de isolados vaginais. Entretanto, não houve nenhuma correlação com os genótipos. Os diferentes isolados expressaram potencial de virulência similares, sugerindo que outros fatores relacionados às leveduras e ao hospedeiro devem participar no desenvolvimento da doença clínica.

[Microbiological identification and in vitro sensitivity of Candida isolates from the oral cavity of HIV-positive individuals]. / Identificação microbiológica e sensibilidade in vitro de Candida isoladas da cavidade oral de indivíduos HIV positivos.

Oropharyngeal candidiasis is the most common fungal infection among patients infected with the human immunodeficiency virus. It can be treated with either systemic or topical antifungal agents, which are indicated empirically on the basis of clinical data. The objective of this study was to determine the frequency of yeast in mouthwashes from HIV-positive patients, compare the results between patients presenting different states of immunodeficiency, and investigate the susceptibility profile of the species isolated in relation to antifungal agents, with the aim of evaluating whether the treatments used in clinical practice are able to reach the majority of the species identified. Yeasts were isolated from 58% of the mouthwash samples collected. Candida albicans was the most (93%) frequent species. Resistance or dose-dependent susceptibility in relation to the antifungal agent tested was registered in approximately 17% of the samples. The significant variability of responses suggests that there are limitations regarding the effectiveness of the empirical therapies instituted.

Fusarium spp. as agents of onychomycosis in immunocompetent hosts.

BACKGROUND: Fusarium spp. are nondermatophyte filamentous fungi, frequently reported as an etiologic agent of opportunistic infections in humans; however, their involvement in the etiology of cutaneous lesions is still debatable, especially in immunocompetent patients, where they are often considered as contaminant fungi. OBJECTIVE: The aims of this study were to report the high prevalence of onychomycosis by Fusarium spp. in immunocompetent patients in the region of Maringá, Paraná, Brazil, to establish clinical and laboratory criteria for this genus as a causal agent of onychomycosis, and to determine the susceptibility profile to the systemic antifungal drugs most frequently used in Brazil (itraconazole, ketoconazole, terbinafine, and amphotericin B). METHODS: The fungi were isolated and identified through the classical method, and sensitivity tests were carried out according to the National Committee for Clinical Laboratory Standards (NCCLS) M38-A protocol. RESULTS: Of the 360 confirmed cases of onychomycosis, 27 (7.5%) were attributed to the genus Fusarium, and F. oxysporum was the most commonly isolated species. Nail lesions with paronychia and pain, combined with direct suggestive microscopy and a high concentration of microorganisms, were predictive of onychomycosis by Fusarium spp. The minimum inhibitory concentration was high for itraconazole, ketoconazole, and terbinafine, but low for amphotericin B. CONCLUSIONS: It is recommended that more attention should be given to the interpretation and identification of species of the Fusarium genus in superficial clinical samples. This fungus may be considered as an agent of onychomycosis, even in immunocompetent individuals, by identifying criteria that separate situations of clinical significance from those of simple contamination.

Identificação microbiológica e sensibilidade in vitro de Candida isoladas da cavidade oral de indivíduos HIV positivos / Microbiological identification and in vitro sensitivity of Candida isolates from the oral cavity of HIV-positive individuals

A candidíase orofaríngea é a infecção fúngica mais comum entre os pacientes infectados pelo vírus da imunodeficiência humana e seu tratamento é realizado com antifúngicos tópicos ou sistêmicos, que são indicados empiricamente com base em dados clínicos. O objetivo deste estudo foi determinar a freqüência de leveduras em lavados bucais de indivíduos HIV positivos, comparando os resultados entre pacientes com diferentes condições de imunodeficiência e verificar o perfil de susceptibilidade das espécies isoladas frente aos antifúngicos visando avaliar se as opções de tratamento utilizadas na prática clínica atingem a maioria das espécies identificadas. Leveduras foram isoladas em 58 por cento das amostras de lavado bucal coletadas e Candida albicans foi a espécie mais (93 por cento) freqüente. Resistência ou susceptibilidade dose dependente, frente aos antifúngicos testados foi registrada em aproximadamente 17 por cento das amostras. A importante variabilidade de resposta sugere limitações quanto à eficácia das terapias instituídas empiricamente.

Oropharyngeal candidiasis is the most common fungal infection among patients infected with the human immunodeficiency virus. It can be treated with either systemic or topical antifungal agents, which are indicated empirically on the basis of clinical data. The objective of this study was to determine the frequency of yeast in mouthwashes from HIV-positive patients, compare the results between patients presenting different states of immunodeficiency, and investigate the susceptibility profile of the species isolated in relation to antifungal agents, with the aim of evaluating whether the treatments used in clinical practice are able to reach the majority of the species identified. Yeasts were isolated from 58 percent of the mouthwash samples collected. Candida albicans was the most (93 percent) frequent species. Resistance or dose-dependent susceptibility in relation to the antifungal agent tested was registered in approximately 17 percent of the samples. The significant variability of responses suggests that there are limitations regarding the effectiveness of the empirical therapies instituted.

Antifungal activity of propolis extract against yeasts isolated from onychomycosis lesions.

The aim of this study was to determine the in vitro activity of propolis extract against 67 yeasts isolated from onychomycosis in patients attending at the Teaching and Research Laboratory of Clinical Analysis of the State University of Maringá. The method used was an adaptation made from the protocol approved by the National Committee for Clinical Laboratory Standards. The yeasts tested were: Candida parapsilosis 35%, C. tropicalis 23%, C. albicans 13%, and other species 29%. The propolis extract showed excellent performance regarding its antifungal activity: the concentration capable of inhibiting the all of the yeasts was 5 x 10(-2) mg/ml of flavonoids and 2 x 10(-2) mg/ml of flavonoids stimulated their cellular death. Trichosporon sp. were the most sensitive species, showing MIC50 and MIC90 of 1.25 x 10(-2) mg/ml of flavonoids, and C. tropicalis was the most resistant, with CFM50 of 5 x 10(-2) mg/ml of flavonoids and MFC90 of 10 x 10(-2) mg/ml. In view of the fact that propolis is a natural, low cost, nontoxic product with proven antifungal activity, it should be considered as another option in the onychomycosis treatment.

Antifungal activity of propolis extract against yeasts isolated from onychomycosis lesions

The aim of this study was to determine the in vitro activity of propolis extract against 67 yeasts isolated from onychomycosis in patients attending at the Teaching and Research Laboratory of Clinical Analysis of the State University of Maringá. The method used was an adaptation made from the protocol approved by the National Committee for Clinical Laboratory Standards. The yeasts tested were: Candida parapsilosis 35 percent, C. tropicalis 23 percent, C. albicans 13 percent, and other species 29 percent. The propolis extract showed excellent performance regarding its antifungal activity: the concentration capable of inhibiting the all of the yeasts was 5 Î 10-2 mg/ml of flavonoids and 2 Î 10-2 mg/ml of flavonoids stimulated their cellular death. Trichosporon sp. were the most sensitive species, showing MIC50 and MIC90 of 1.25 Î 10-2 mg/ml of flavonoids, and C. tropicalis was the most resistant, with CFM50 of 5 Î 10-2 mg/ml of flavonoids and MFC90 of 10 Î 10-2 mg/ml. In view of the fact that propolis is a natural, low cost, non-toxic product with proven antifungal activity, it should be considered as another option in the onychomycosis treatment.

Caracterização de leveduras isoladas da vagina e sua associação com candidíase vulvovaginal em duas cidades do sul do Brasil / Characterization of yeasts isolated from the vagina and their association with vulvovaginal candidíasis in two cities of the South of Brazil

OBJETIVOS: avaliar a distribuição de espécies de leveduras isoladas da vagina em duas localidades do sul do Brasil e comparar o perfil de suscetibilidade in vitro destas leveduras a antifúngicos usados na prática clínica. MÉTODOS: todas as mulheres atendidas entre janeiro e junho de 2004 para exames rotineiros de amostras vaginais, independente de serem sintomáticas ou não, foram incluídas neste estudo. Foram excluídas as que apresentavam imunodeficiências como AIDS ou outras infecções genitais. Amostras de conteúdo vaginal dessas mulheres (Jaraguá do Sul - SC (n=130) e Maringá - PR (n=97)) foram cultivadas. As leveduras isoladas foram identificadas e submetidas ao teste de suscetibilidade aos antifúngicos fluconazol, nistatina e anfotericina B. RESULTADOS: a freqüência de cultura positiva para levedura foi semelhante nas duas localidades, aproximadamente 24 por cento. Candida albicans foi a espécie prevalente, mas sua freqüência diferiu: em SC correspondeu a 77,4 por cento das leveduras e foi a mais freqüente tanto nas mulheres sintomáticas quanto nas assintomáticas. Já no PR foi 50,0 por cento, com predomínio mais evidente nos casos sintomáticos. Observamos altos índices de suscetibilidade ao fluconazol e anfotericina B, porém 51,1 por cento das leveduras apresentaram suscetibilidade dependente da dose (S-DD) para nistatina. C. albicans mostrou maior tendência de resistência à nistatina (52,8 por cento de S-DD) do que as espécies não-albicans (44,4 por cento). CONCLUSÕES: nossos dados mostram diferenças regionais quanto à espécie de levedura em amostras vaginais. Sugerem que a determinação da espécie pode ter implicação clínica, considerando as diferenças quanto à suscetibilidade, principalmente à nistatina, e que poderiam ter importância no manejo da candidíase vulvovaginal