Comparison of in vitro activity of five antifungal agents against dermatophytes, using the agar dilution and broth microdilution methods.

The purpose of this study was to compare the agar dilution and broth microdilution methods for determining the minimum inhibitory concentration (MIC) of fluconazole, itraconazole, ketoconazole, griseofulvin and terbinafine for 60 dermatophyte samples belonging to the species Trichophyton rubrum, Trichophyton mentagrophytes and Microsporum canis. The percentage agreement between the two methods, for all the isolates with < 2 dilutions that were tested was 91.6% for ketoconazole and griseofulvin, 88.3% for itraconazole, 81.6% for terbinafine and 73.3% for fluconazole. One hundred percent agreement was obtained for Trichophyton mentagrophytes isolates evaluated with ketoconazole and griseofulvin. Thus, until a reference method for testing the in vitro susceptibility of dermatophytes is standardized, the similarity of the results between the two methods means that the agar dilution method may be useful for susceptibility testing on these filamentous fungi.

Comparison of in vitro activity of five antifungal agents against dermatophytes, using the agar dilution and broth microdilution methods / Comparação da atividade in vitro de cinco agentes antifúngicos para dermatófitos, usando os métodos de diluição em ágar e microdiluição em caldo

The purpose of this study was to compare the agar dilution and broth microdilution methods for determining the minimum inhibitory concentration (MIC) of fluconazole, itraconazole, ketoconazole, griseofulvin and terbinafine for 60 dermatophyte samples belonging to the species Trichophyton rubrum, Trichophyton mentagrophytes and Microsporum canis. The percentage agreement between the two methods, for all the isolates with < 2 dilutions that were tested was 91.6 percent for ketoconazole and griseofulvin, 88.3 percent for itraconazole, 81.6 percent for terbinafine and 73.3 percent for fluconazole. One hundred percent agreement was obtained for Trichophyton mentagrophytes isolates evaluated with ketoconazole and griseofulvin. Thus, until a reference method for testing the in vitro susceptibility of dermatophytes is standardized, the similarity of the results between the two methods means that the agar dilution method may be useful for susceptibility testing on these filamentous fungi.

O propósito do presente trabalho foi comparar os métodos de diluição em ágar e diluição em caldo para a determinação de concentração inibitória mínima de fluconazol, itraconazol, cetoconazol, griseofulvina e terbinafina para 60 amostras de dermatófitos pertencentes às espécies, Trichophyton rubrum, Trichophyton. mentagrophytes e Microsporum canis. A porcentagem de acordo entre os dois métodos para todos os isolados testados considerando-se valores < 2 diluições, foram de 91,6 por cento para cetoconazol e para griseofulvina, de 88,3 por cento para itraconazol, de 81,6 por cento para terbinafina e de 73,3 por cento para fluconazol. Uma concordância de 100 por cento foi obtido para isolados de Trichophyton mentagrophytes avaliados com cetoconazol e griseofulvina. Desta forma, até que um método de referência seja padronizado para testar a suscetibilidade in vitro para os dermatófitos, os resultados semelhantes encontrados para os dois métodos fazem com que o método de diluição em ágar possa ser útil no teste de suscetibilidade para estes fungos filamentosos.

In vitro susceptibility testing of dermatophytes isolated in Goiania, Brazil, against five antifungal agents by broth microdilution method.

The antifungal activities of fluconazole, itraconazole, ketoconazole, terbinafine and griseofulvin were tested by broth microdilution technique, against 60 dermatophytes isolated from nail or skin specimens from Goiania city patients, Brazil. In this study, the microtiter plates were incubated at 28 masculineC allowing a reading of the minimal inhibitory concentration (MIC) after four days of incubation for Trichophyton mentagrophytes and five days for T. rubrum and Microsporum canis. Most of the dermatophytes had uniform patterns of susceptibility to the antifungal agents tested. Low MIC values as 0.03 microg/mL were found for 33.3%, 31.6% and 15% of isolates for itraconazole, ketoconazole and terbinafine, respectively.

In vitro susceptibility testing of dermatophytes isolated in Goiania, Brazil, against five antifungal agents by broth microdilution method / Teste de suscetibilidade in vitro de dermatófitos isolados em Goiânia, Brasil, contra cinco agentes antifúngicos pelo método de microdiluição em caldo

The antifungal activities of fluconazole, itraconazole, ketoconazole, terbinafine and griseofulvin were tested by broth microdilution technique, against 60 dermatophytes isolated from nail or skin specimens from Goiania city patients, Brazil. In this study, the microtiter plates were incubated at 28 ºC allowing a reading of the minimal inhibitory concentration (MIC) after four days of incubation for Trichophyton mentagrophytes and five days for T. rubrum and Microsporum canis. Most of the dermatophytes had uniform patterns of susceptibility to the antifungal agents tested. Low MIC values as 0.03 µg/mL were found for 33.3 percent, 31.6 percent and 15 percent of isolates for itraconazole, ketoconazole and terbinafine, respectively.

Atividades antifúngicas de fluconazol, itraconazol, cetoconazol, terbinafina e griseofulvina foram testadas pelo método de microdiluição em caldo contra 60 isolados de dermatófitos. Os resultados mostraram que todos os isolados produziram crescimento claramente detectável a 28 ºC e a concentração inibitória mínima (CIM) foi determinada após quatro dias de incubação para Trichophyton mentagrophytes e cinco dias para T. rubrum e Microsporum canis. A maioria dos isolados teve um padrão uniforme de suscetibilidade para os agentes antifúngicos testados. Baixos valores de CIM como 0,03 µg/mL foram encontrados para 33,3 por cento, 31,6 por cento e 15 por cento dos isolados para itraconazol, cetoconazol e terbinafina, respectivamente.

Antifungal activities of azole agents against the Malassezia species.

In this paper, we identified 95 Malassezia isolates by morphological and biochemical criteria and assessed the in vitro activity of fluconazole, itraconazole, ketoconazole and voriconazole by broth microdilution against these species using slightly modified Leeming-Notman medium. The Malassezia isolates were identified as M. furfur (74), M. sympodialis (11), M. obtusa (8) and M. globosa (2). The modified Leeming-Notman medium used for susceptibility testing allowed good growth of Malassezia spp. Visual reading of the minimal inhibitory concentration (MIC) was readily achieved until Day 5 of incubation at 32 degrees C. Although high MIC values of 16 microg/mL for fluconazole were observed in 9.5% of Malassezia isolates, in general these microorganisms were susceptible to all drugs studied. Interestingly, one M. globosa isolate showed high MIC values for voriconazole, itraconazole and fluconazole. For the 95 strains, the MIC ranges were <0.03-4 microg/mL for ketoconazole, <0.03 to >16 microg/mL for voriconazole, <0.125 to >64 microg/mL for fluconazole and <0.03-16 microg/mL for itraconazole. In summary, the good reproducibility and visual readings obtained using modified Leeming-Notman medium suggest that this medium should be proposed for antifungal testing of drugs against Malassezia spp.

Identificação de espécies de Malassezia em pacientes com pitiríase versicolor em Goiânia-GO / Identification of Malassezia species in patients with pityriasis versicolor in Goiânia-GO

O objetivo deste estudo foi verificar a freqüência de pitiríase versicolor e identificar leveduras do gênero Malassezia, de pacientes encaminhados ao laboratório de Micologia da Universidade Federal de Goiás em Goiânia. Foram diagnosticados 95 casos de pitíriase versicolor e identificados quatro espécies de Malassezia: Malassezia furfur, Malassezia sympodialis, Malassezia globosa e Malassezia obtusa.

The objectives of this study were to investigate the frequency of pityriasis versicolor occurrence and to identify yeasts of the genus Malassezia in patients at the mycology laboratory of the Federal University of Goiás, in Goiânia, State of Goiás. Ninety-five cases of pityriasis versicolor were diagnosed, and four species of Malassezia were identified: Malassezia furfur, Malassezia sympodialis, Malassezia globosa and Malassezia obtusa.

Asymptomatic oral carriage of Candida species in HIV-infected patients in the highly active antiretroviral therapy era.

Oropharyngeal candidiasis is the most common opportunistic fungal infection in individuals infected with human immunodeficiency virus. CD4+ lymphocytes count and the quantification of viral RNA in blood plasma have been found to be the main markers of HIV disease progression. The present study was conducted to evaluate Candida sp. diversity in the oral cavity of HIV-infected patients and to determine whether there was association of CD4+ cell count and viral load with asymptomatic oral Candida carriage. Out of 99 HIV-positive patients studied, 62 (62.6%) had positive culture for Candida (oral carriage) and 37 patients (37.4%) had Candida negative culture (no oral carriage). The etiologic agents most common were C. albicans and C. tropicalis. The range of CD4+ was 6-2305 cells/mm3 in colonized patients and 3-839 cells/mm3 for non-colonized patients, while the viral load was 60-90016 copies/mL for colonized patients and 75-110488 copies/mL for non colonized patients. The viral load was undetectable in 15 colonized patients and in 12 non colonized patients. Our results showed that there was no significant difference of the variables CD4+ cell count and viral load between oral candida carriage and no oral candida carriage patients.

Asymptomatic oral carriage of Candida species in HIV-infected patients in the highly active antiretroviral therapy era / Carreadores assintomáticos de espécies de Candida na mucosa bucal de pacientes infectados pelo HIV na era da terapia antiretroviral

Oropharyngeal candidiasis is the most common opportunistic fungal infection in individuals infected with human immunodeficiency virus. CD4+ lymphocytes count and the quantification of viral RNA in blood plasma have been found to be the main markers of HIV disease progression. The present study was conducted to evaluate Candida sp. diversity in the oral cavity of HIV-infected patients and to determine whether there was association of CD4+ cell count and viral load with asymptomatic oral Candida carriage. Out of 99 HIV-positive patients studied, 62 (62.6 percent) had positive culture for Candida (oral carriage) and 37 patients (37.4 percent) had Candida negative culture (no oral carriage). The etiologic agents most common were C. albicans and C. tropicalis. The range of CD4+ was 6-2305 cells/mm³ in colonized patients and 3-839 cells/mm³ for non-colonized patients, while the viral load was 60-90016 copies/mL for colonized patients and 75-110488 copies/mL for non colonized patients. The viral load was undetectable in 15 colonized patients and in 12 non colonized patients. Our results showed that there was no significant difference of the variables CD4+ cell count and viral load between oral candida carriage and no oral candida carriage patients.

Candidíase de orofaringe é a infecção fúngica oportunística mais comum em indivíduos infectados com o vírus da imunodeficiência humana. Contagem de linfócitos CD4+ e quantificação de RNA viral no plasma sanguíneo são os principais marcadores da progressão da doença pelo HIV. O presente estudo foi conduzido para avaliar a diversidade de espécies de Candida presentes na cavidade bucal de pacientes infectados pelo HIV e para determinar se havia associação de contagem de células CD4+ e de carga viral com carreadores assintomáticos de Candida, na mucosa bucal. Dos 99 pacientes HIV positivo estudados, 62 (62,6 por cento) apresentaram cultura positiva para Candida sp. sendo denominados carreadores de Candida e os 37 pacientes (37,4 por cento) que não possuíam leveduras do gênero Candida na mucosa bucal foram denominados não carreadores. Os agentes etiológicos mais comuns foram C. albicans e C. tropicalis. A variação de CD4+ foi de 6-2305 cels/mm³ em pacientes colonizados e de 3-839 cels/mm³ para pacientes não colonizados, enquanto a carga viral variou de 60-90016 cópias/mL para pacientes colonizados e de 75-110488 cópias/mL para não colonizados. Não foi possível a detecção de carga viral em 15 pacientes colonizados e em 12 não colonizados, porque o limite mínimo de detecção era de 50 cópias/mL. Nossos resultados mostraram que não houve diferença significante na contagem de células CD4+ e de carga viral entre os pacientes carreadores e não carreadores de Candida na mucosa bucal de pacientes com AIDS.

[Identification of Malassezia species in patients with pityriasis versicolor in Goiânia-GO]. / Identificação de espécies de Malassezia em pacientes com pitiríase versicolor em Goiânia-GO.

The objectives of this study were to investigate the frequency of pityriasis versicolor occurrence and to identify yeasts of the genus Malassezia in patients at the mycology laboratory of the Federal University of Goiás, in Goiânia, State of Goiás. Ninety-five cases of pityriasis versicolor were diagnosed, and four species of Malassezia were identified: Malassezia furfur, Malassezia sympodialis, Malassezia globosa and Malassezia obtusa.

Candida colonization in intensive care unit patients' urine

The objective of this study was to identify possible predisposing factors for candiduria in intensive care unit (ICU) patients from Hospital das Clínicas, Universidade Federal de Goiás, Goiânia, Brazil, during one year. Urine samples from 153 ICU patients were obtained by catheterization on admission day and every seven days. Data such as sex, age, antifungal therapy, and variables as antibiotics, underlying diseases or comorbid conditions and stay in the hospital, were collected from patients who had at least one urine culture that yielded > 10³ yeast colonies/ml. Candiduria was recovered in 68 patients and the commonest predisposing factors were antibiotic therapy (100 percent) and indwelling urinary catheter (92.6 percent). The percentage of Candida spp. isolation increased during the extended periods in which patients remained in the ICU. C. albicans was isolated in 69.1 percent, and the other species non-albicans as C. glabrata, C. kefyr, C. parapsilosis, C. famata, C. guilliermondii, C. krusei, and C. tropicalis were isolated in lower percentage. The high frequency of candiduria and the possible predisposing factors found in ICU patients show that candiduria surveillance should be performed to help reducing nosocomial infections.

Identificação de leveduras do gênero Candida nas unhas e em descamação de pele em Goiânia (GO), durante o ano de 2003 / Identification of yeasts of genus Candida on nails and skin desquamation in Goiania (GO), during the year 2003

A candidíase pode ser descrita como a mais freqüente infecção fúngica oportunística. Ela produz lesões que variam de cutâneas a sistêmicas. Embora a Candida albicans seja o agente mais comumente isolado, espécies como C. tropicalis, C. parapsilosis, C. guilliermondii e C. krusei têm sido identificadas como agentes de candidíase. Neste trabalho, foram coletados materiais de lesões superficiais de diferentes regiões do corpo de 2.083 pacientes do Laboratório de Micologia IPTSP-UFG, durante o ano de 2003, para identificação de fungos do gênero Candida. As amostras coletadas foram submetidas a exame direto, usando-se KOH a 40por cento, e cultivadas em ágar Sabouraud dextrose acrescido de cloranfenicol. Os isolados foram identificados por produção de tubo germinativo e clamídoconídios e assimilação de hidratos de carbono. Foram isoladas 190 leveduras do gênero Candida (9,1por cento), entre as quais C. albicans (63,2por cento), C. parapsilosis (14,2por cento) e C. tropicalis (9,5por cento) foram as espécies mais freqüentes. As lesões por Candida foram predominantes nas unhas das mãos (42,1por cento) e dos pés (42,6por cento). Os resultados obtidos mostraram um aumento de C. parapsilosis e C. tropicalis como agentes de candidíase, comprovando a participação de outras espécies não-albicans como patógenos emergentes.

Identificação das leveduras do gênero Candida por métodos manuais convencionais e pelo método cromógeno CHROMagar(TM) Candida / Identification of Candida species by conventional methods and by chromogenic medium CHROMagar(TM) Candida

Diferentes meios cromógenos de cultivo, fundamentados na cor desenvolvida pelas colônias através de indicadores de pH e fermentação de compostos específicos ou substratos cromógenos, têm sido utilizados para diferenciar Candida albicans e outras leveduras de interesse clínico. O meio de cultura CHROMagar Candida é usado para isolar e identificar presuntivamente C. albicans, C. krusei e C. tropicalis. Neste trabalho, o meio cromógeno CHROMagar(TM) Candida foi usado para a identificação de 53 leveduras previamente identificadas por métodos manuais convencionais como Candida albicans(34), C.tropicalis(7), C.krusei(5) C.glabrata(4) e C.parapsilosis(3). Das 34 amostras identificadas por métodos convencionais como C. albicans, 30 (88,2por cento) apresentaram a coloração verde-escura brilhante própria desta espécie no meio cromogênico. As características relativas à coloração e à morfologia, mostradas em CHROMagar Candida, apresentaram-se compatíveis com C. krusei, C. glabrata e C. parapsilosis em todas as amostras estudadas, enquanto 5 dos 7 isolados (71,4por cento) de C. tropicalis apresentaram a coloração azul característica desta espécie. Com os resultados obtidos concluiu-se que o meio CHROMagar Candida pode ser útil na identificação presuntiva das espécies do gênero Candida.

Candida colonization in intensive care unit patients' urine.

The objective of this study was to identify possible predisposing factors for candiduria in intensive care unit (ICU) patients from Hospital das Clínicas, Universidade Federal de Goiás, Goiânia, Brazil, during one year. Urine samples from 153 ICU patients were obtained by catheterization on admission day and every seven days. Data such as sex, age, antifungal therapy, and variables as antibiotics, underlying diseases or comorbid conditions and stay in the hospital, were collected from patients who had at least one urine culture that yielded > 10(3) yeast colonies/ml. Candiduria was recovered in 68 patients and the commonest predisposing factors were antibiotic therapy (100%) and indwelling urinary catheter (92.6%). The percentage of Candida spp. isolation increased during the extended periods in which patients remained in the ICU. C. albicans was isolated in 69.1%, and the other species non-albicans as C. glabrata, C. kefyr, C. parapsilosis, C. famata, C. guilliermondii, C. krusei, and C. tropicalis were isolated in lower percentage. The high frequency of candiduria and the possible predisposing factors found in ICU patients show that candiduria surveillance should be performed to help reducing nosocomial infections.

Candiduria in hospital patients: a study prospective.

The presence of Candida species in the urine is frequent among hospitalized patients. We studied sample urine of 205 hospitalized patients during a 1-year period to determine the incidence of nosocomial candiduria. The yeasts were isolated in 22% (45/205) urine cultures and risk factors in these patients were analyzed. Candida albicans was isolated in 35.6% and C. tropicalis (22%) was the second most frequent species isolated. Most patients were women (57.8%) with a mean age of 48.7 years. The principal risk factors that were observed in patients with candiduria included antibiotics therapy (100%), urinary catheterization (84.4%), surgical procedure (66.7%), female sex and extended hospitalization. The efficacy of fluconazole therapy to eradicate Candida from urine was demonstrated (p =0.05). Of the 23 individuals who received antifungal therapy, candiduria persisted in 9 (39.2%) and of 22 patients who received no antifungal therapy, the candiduria persisted in 15 (68.2%).