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Master's Dissertation
DOI
https://doi.org/10.11606/D.9.2019.tde-14102019-114206
Document
Author
Full name
Tânia Sueli de Andrade
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 1999
Supervisor
Committee
Cury, Arlete Emily (President)
Mamizuka, Elsa Masae
Melhem, Marcia de Souza Carvalho
Title in Portuguese
Suscetibilidade in vitro de agentes causadores de cromoblastomicose frente a diferentes antifúngicos
Keywords in Portuguese
Anfotericina B
Antifúngicos (Farmacologia)
Antifungigrama
Cromoblastomicose
Fonsecaea pedrosoi
Itraconazol
Micologia
Terbinafina
Abstract in Portuguese
Diversos recursos terapêuticos são utilizados para controle ou cura da cromoblastomicose, mas ainda não existe terapia padrão. Poucos são os estudos relacionados à resposta dos fungos causadores frente a antifúngicos atualmente disponíveis. Nosso objetivo foi determinar os perfis de suscetibilidade desses agentes aos diferentes antifúngicos utilizados no tratamento das micoses, através da padronização da técnica de diluição em ágar e correlacionar com o tratamento vigente para alguns pacientes. Foram estudadas 39 cepas de agentes de cromoblastomicose, sendo, 12 de coleções; 30 foram identificadas como Fonsecaea pedrosoi, 6 como Phialophora verrucosa e cada uma das outras como: Cladophialophora (Cladosporium) carrionii, Fonsecaea compacta e Rhinocladiella aquaspersa. Foram realizados estudos preeliminares para escolha das melhores condições para os testes de suscetibilidade. O meio selecionado foi o yeast nitrogen base tamponado com fosfato (YNBP). O inóculo contendo cerca de 4 x 104 células/ml foi preparado a partir de cepas com 14 dias de desenvolvimento. A incubação foi realizada a 30°C e a leitura ocorreu do 3° ao 7°dias de incubação. Os testes foram realizados com: anfotericina B nas concentrações de 0,25µg/ml a 64µg/ml, 5-fluorocitosina de 0,3µg/ml a 100µg/ml, fluconazol de 0,25µg/ml a 128µg/ml, terbinafina de 0,09µg/ml a 24µg/ml, itraconazol e cetoconazol de 0,125µg/ml a 32µg/ml. Para auxiliar na interpretação dos resultados, o estabelecimento de sensiblidade ou resistência foi baseado nos níveis séricos alcançados pelas drogas. Apenas uma cepa foi sensível no teste de CIM a anfotericina B. Os valores mais altos para anfotericina B estavam entre os pacientes que fazem uso de itraconazol. Entre as 39 cepas estudadas, 77% foram resistentes ao itraconazol no teste de CIM. Para as cepas de 4 pacientes isoladas de amostras seriadas houve aumento das C IMs e para dois deles não houve melhora clínica com itraconazol. Em um desses casos houve resistência cruzada com fluconazol e outro com cetoconazol e anfotericina B. Todas as cepas foram sensíveis ao cetoconazol, mas resistentes ao fluconazol nos testes de CIM e CFM. Apenas 7 cepas responderam a 5-fluorocitosina no teste de CIM. O cetoconazol foi a droga mais eficaz, seguido da terbinafina, apesar de algumas cepas apresentaram resistência a mesma, principalmente nas procedentes de coleções. Os dados obtidos sugerem resistência inata dos fungos em estudos a algumas drogas e adquirida para algumas cepas de pacientes em tratamento com itraconazol. Esses mecanismos devem ser melhor pesquisados, além da eficácia associação de drogas, ou outra terapia e do melhor desempenho da terbinafina nos testes.
Title in English
Susceptibility "in vitro'' of agents of the chromoblastomycosis against differs antifungals
Keywords in English
Anfotericina B
Antifúngicos (Farmacologia)
Antifungigrama
Cromoblastomicose
Fonsecaea pedrosoi
Itraconazol
Micologia
Terbinafina
Abstract in English
Several therapeutics resources are used to control or cure of chromoblastomycossis, but there is not stand therapy yet. Feel are the studies related with fungi agents against attual antifungals. The aim of this study was to determine susceptibility profiles this fungi against differs antifungals used in treatment of mycosis, through the standaring agar diluition thecnica and relation with treatment to some pacients. 39 strains of agents were studied of chromoblastomycosis, 12 were collections, 30 were identified as F. pedrosoi, 6 as P. verrucosa and each one of others as: C. carrionii. F. compacta and R. aquaspersa. Previous studies were performed to choose the best conditions for susceptibilities tests. The culture medium selected was yeast nitrogen base buffering with phosphate (YNBP). The inoculum about 4x104 cels/ml. Was performed with strains with 14 days of developement. The incubation was performed of 30°C and the reading ocurred from the 3° to the 7° days of incubation. The tests were performed with amphotericin B in concentration of 0,25µg/ml to 64µg/ml, 5-flucytosin from 0,3µg/ml to 100µg/ml, fluconazole from 0,25µg/ml to 128µg/ml, terbinafine from 0,09µg/ml to 24µg/ml, itraconazole and ketoconazole from 0,125µg/ml to 32µg/ml. The interpretation of results was based in the plasma levels reached by the drugs. Only one strain was susceptible in the test of MIC for amphotericin B. The values highest for AmB were among the patients who used itraconazole. Among the 39 strains studied, 77% were resistents to the itraconazole in the test of MIC. For the strains there was increase of the MICs for the strains of four patients recovered from sequential samples and for to of them was not improvement with itraconazole. At one of these cases there was cross resistance with fluconazole and other with ketoconazole and amphotericin B. All the strains were susceptibles to ketoconazole, but resistents to the fluconazole in the MIC's and MFC's tests. Only 7 strains were sensibles to 5-FC in the MIC test. The terbinafine was the best drug, despite of some strains showed resistence, mainly in the culture collection. The results suggest intrinsic resistance of the fungi in study to some drugs and acquired resistance for some strains of the patients in treatment with itraconazole. These mechanism must be more researched efficacy association of drugs, or other therapy and of the best fulfillment of the terbinafine in the tests.
 
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Publishing Date
2019-10-18
 
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