Atividade in vitro de antifúngicos em biofilmes de Candida

Detalhes bibliográficos
Autor(a) principal: Pereira, Núbia Pontes
Data de Publicação: 2011
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Repositório Institucional da UFG
dARK ID: ark:/38995/0013000002h3s
Texto Completo: http://repositorio.bc.ufg.br/tede/handle/tede/9240
Resumo: C. albicans infections may be favored by tissue damages such insertion of medical devices that function as a substrate for fungus growth. This growth can occur in the form of three-dimensional structures formed by yeast and hyphae, coated with a matrix of extracellular polymeric material, forming a biofilm. This agregated of cells is characterized by having greater resistance to host defenses and antifungals, which makes hard treating these infections. This study aimed to determine the kinetics of biofilm formation by isolates of Candida sp. and to evaluate the antifungal susceptibility of plancktonic and sessile cells. We evaluated 10 isolates of C. albicans from different materials, isolated and identified by standardized methods. The minimum inhibitory concentrations (MICs) and minimal fungicidal concentrations (MFCs) of amphotericin B, fluconazole, voriconazole and itraconazole to plancktonic cells were performed according to protocol M27-A3 of Clinical Laboratory Standard Institute (CLSI). Verification of kinetics of biofilms’ formation was assessed at 24 h, 48 h, 72 h and 96 h of incubation at 37 ° C and in vitro susceptibility of sessile cells, was evaluated using the MTT (3 - [4,5-Dimethyl-2-Thiazy] -2,5- Diphenyl-2H-Tetrazolium bromide) assay. The analysis of kinetics at 24 h showed that all isolates produced biofilm and the highest metabolic activity occurred within 48h of growth. Significant differences, on the metabolic activity of biofilms were observed on strains U02, M02 and V60, which produced the highest amount of biofilm with 48 h of growth. MIC values obtained for plancktonic cells ranged from 1 to 8 mg/mL for fluconazole; between 0.0625 to 1 mg/mL for itraconazole; between 0.0312 to 0.25 g/mL for voriconazole; and between 0.25 to 2 mg/mL for amphotericin B. The MFC was twice the MIC for all isolates. For the values obtained for fluconazole the MICS50 ranged from 8 mg/mL to > 1024 mg/mL and MICS80 ≥ 1024 mg/mL. MICS50 for itraconazole ranged from 4 mg/mL to > 16 g/mL and MICS80 ≥ 16 g/mL. The MICS50 for voriconazole showed values between 4 mg/mL and > 16 g/mL and MICS80 ≥ 16 g/mL. Amphotericin B had MICS50 values of 0.25 mg / mL to > 16 g/mL and values MICS80 of 2 mg/mL to > 16 g/mL. We could demonstrate an increased resistance of biofilms to major antifungal drugs by evaluating the susceptibility of C. albicans isolates as plancktonic and sessile cells. These results show the need of new therapeutic or preventive strategies to reduce the mortality rate of patients with infections associated with Candida biofilms.
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spelling Souza, Lúcia Kioko Hasimoto ehttp://lattes.cnpq.br/4557218510118165Souza, Lúcia Kioko Hasimoto eFaganello, JosianeSilva, Maria do Rosário Rodrigueshttp://lattes.cnpq.br/8992795603947194Pereira, Núbia Pontes2019-01-22T10:53:29Z2011-05-16PEREIRA, Núbia Pontes. Atividade in vitro de antifúngicos em biofilmes de Candida. 2011. 67 f. Dissertação (Mestrado em Medicina Tropical e Saúde Publica) - Universidade Federal de Goiás, Goiânia, 2011.http://repositorio.bc.ufg.br/tede/handle/tede/9240ark:/38995/0013000002h3sC. albicans infections may be favored by tissue damages such insertion of medical devices that function as a substrate for fungus growth. This growth can occur in the form of three-dimensional structures formed by yeast and hyphae, coated with a matrix of extracellular polymeric material, forming a biofilm. This agregated of cells is characterized by having greater resistance to host defenses and antifungals, which makes hard treating these infections. This study aimed to determine the kinetics of biofilm formation by isolates of Candida sp. and to evaluate the antifungal susceptibility of plancktonic and sessile cells. We evaluated 10 isolates of C. albicans from different materials, isolated and identified by standardized methods. The minimum inhibitory concentrations (MICs) and minimal fungicidal concentrations (MFCs) of amphotericin B, fluconazole, voriconazole and itraconazole to plancktonic cells were performed according to protocol M27-A3 of Clinical Laboratory Standard Institute (CLSI). Verification of kinetics of biofilms’ formation was assessed at 24 h, 48 h, 72 h and 96 h of incubation at 37 ° C and in vitro susceptibility of sessile cells, was evaluated using the MTT (3 - [4,5-Dimethyl-2-Thiazy] -2,5- Diphenyl-2H-Tetrazolium bromide) assay. The analysis of kinetics at 24 h showed that all isolates produced biofilm and the highest metabolic activity occurred within 48h of growth. Significant differences, on the metabolic activity of biofilms were observed on strains U02, M02 and V60, which produced the highest amount of biofilm with 48 h of growth. MIC values obtained for plancktonic cells ranged from 1 to 8 mg/mL for fluconazole; between 0.0625 to 1 mg/mL for itraconazole; between 0.0312 to 0.25 g/mL for voriconazole; and between 0.25 to 2 mg/mL for amphotericin B. The MFC was twice the MIC for all isolates. For the values obtained for fluconazole the MICS50 ranged from 8 mg/mL to > 1024 mg/mL and MICS80 ≥ 1024 mg/mL. MICS50 for itraconazole ranged from 4 mg/mL to > 16 g/mL and MICS80 ≥ 16 g/mL. The MICS50 for voriconazole showed values between 4 mg/mL and > 16 g/mL and MICS80 ≥ 16 g/mL. Amphotericin B had MICS50 values of 0.25 mg / mL to > 16 g/mL and values MICS80 of 2 mg/mL to > 16 g/mL. We could demonstrate an increased resistance of biofilms to major antifungal drugs by evaluating the susceptibility of C. albicans isolates as plancktonic and sessile cells. These results show the need of new therapeutic or preventive strategies to reduce the mortality rate of patients with infections associated with Candida biofilms.Infecções por C. albicans podem ser favorecidas por danos causados aos tecidos, como na inserção de dispositivos médicos, que funcionam como substrato para o crescimento do fungo. Este crescimento pode ocorrer na forma de estruturas tridimensionais, formadas por leveduras e hifas, revestidas por uma matriz de material polimérico extracelular, configurando um biofilme. Este agregado de células caracteriza-se por apresentar maior resistência às defesas do hospedeiro e aos antifúngicos, dificultando o tratamento destas infecções. Este trabalho teve como objetivo verificar a cinética de formação de biofilmes por isolados de Candida, e avaliar a suscetibilidade antifúngica de suas células planquitônicas e sésseis. Foram avaliados 10 isolados de C. albicans, provenientes de diferentes materiais, isolados e identificados por métodos padronizados. As concentrações inibitórias mínimas (CIMs) e concentração fungicida mínima (CFM) para anfotericina B, fluconazol, voriconazol e itraconazol de células planquitônicas foram realizadas segundo o protocolo M27-A3 do Clinical Laboratory Standard Institute (CLSI). A verificação da cinética de formação dos biofilmes foi avaliada em 24 h, 48 h, 72 h e 96 h de incubação a 37oC e a suscetibilidade in vitro das células sésseis, foi avaliada utilizando-se o ensaio de MTT (3-[4,5-Dimethyl-2-Thiazyl]-2,5-Diphenyl-2H-Tetrazolium bromide). A análise da cinética de formação mostrou que com 24 h todos os isolados formaram biofilme e, a maior atividade metabólica ocorreu com 48h de crescimento. Diferenças significativas, quanto à atividade metabólica dos biofilmes, foram observadas, sendo os isolados U02, M02 e V60 os que produziram mais biofilme com 48 h de crescimento. Os valores de CIM das células plaquitônicas (CIMP) obtidos para fluconazol variaram de 1 a 8 μg/mL; para itraconazol entre 0,0625 a 1 μg/mL; para voriconazol entre 0,0312 a 0,25 μg/mL e para anfotericina B de 0,25 a 2 μg/mL. A CFM apresentada por todos os isolados avaliados foi o dobro do obtido para a CIM. Para fluconazol os valores de concentração inibitória mínima que reduzirma em 50% a atividade metabólica das células sésseis (CIMS50), variaram de 8 μg/mL a > 1.024 μg/mL e foi ≥ 1.024 μg/mL quando observada uma inibição de 80% da atividade metabólica destas células (CIMS80). Com itraconazol a CIMS50 variou de 4 μg/mL a > 16 μg/mL e a CIMS80 ≥16 μg/mL. A CIMS50 de voriconazol mostrou valores de 4 μg/mL a > 16 μg/mL e a CIMS80 ≥16 μg/mL. Anfotericina B apresentou valores de CIMS50 de 0,25 μg/mL a > 16 μg/mL e valores de CIMS80 de 2 μg/mL a > 16 μg/mL. Através da avaliação da suscetibilidade de isolados de C. albicans, sob a forma de células planquitônicas e biofilmes, foi possível demonstrar um aumento da resistência dos isolados aos principais antifúngicos, tornando necessárias novas estratégias terapêuticas ou preventivas a fim de diminuir a taxa de mortalidade de pacientes com infecções associadas aos biofilmes de Candida.Submitted by Ana Caroline Costa (ana_caroline212@hotmail.com) on 2019-01-21T17:59:55Z No. of bitstreams: 2 Dissertação - Núbia Pontes Pereira - 2011.pdf: 2541686 bytes, checksum: 78b3a93f16f2f902fdbea39491f62fe2 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5)Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2019-01-22T10:53:29Z (GMT) No. of bitstreams: 2 Dissertação - Núbia Pontes Pereira - 2011.pdf: 2541686 bytes, checksum: 78b3a93f16f2f902fdbea39491f62fe2 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5)Made available in DSpace on 2019-01-22T10:53:29Z (GMT). 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dc.title.eng.fl_str_mv Atividade in vitro de antifúngicos em biofilmes de Candida
dc.title.alternative.eng.fl_str_mv In vitro activity of antifungals in Candida biofilms
title Atividade in vitro de antifúngicos em biofilmes de Candida
spellingShingle Atividade in vitro de antifúngicos em biofilmes de Candida
Pereira, Núbia Pontes
Candida
Biofilme
Suscetibilidade
Antifúngicos
Infecções
Biofilm
Susceptibility
Antifungals
Infections
BIOLOGIA E FISIOLOGIA DOS MICROORGANISMOS::MICOLOGIA
title_short Atividade in vitro de antifúngicos em biofilmes de Candida
title_full Atividade in vitro de antifúngicos em biofilmes de Candida
title_fullStr Atividade in vitro de antifúngicos em biofilmes de Candida
title_full_unstemmed Atividade in vitro de antifúngicos em biofilmes de Candida
title_sort Atividade in vitro de antifúngicos em biofilmes de Candida
author Pereira, Núbia Pontes
author_facet Pereira, Núbia Pontes
author_role author
dc.contributor.advisor1.fl_str_mv Souza, Lúcia Kioko Hasimoto e
dc.contributor.advisor1Lattes.fl_str_mv http://lattes.cnpq.br/4557218510118165
dc.contributor.referee1.fl_str_mv Souza, Lúcia Kioko Hasimoto e
dc.contributor.referee2.fl_str_mv Faganello, Josiane
dc.contributor.referee3.fl_str_mv Silva, Maria do Rosário Rodrigues
dc.contributor.authorLattes.fl_str_mv http://lattes.cnpq.br/8992795603947194
dc.contributor.author.fl_str_mv Pereira, Núbia Pontes
contributor_str_mv Souza, Lúcia Kioko Hasimoto e
Souza, Lúcia Kioko Hasimoto e
Faganello, Josiane
Silva, Maria do Rosário Rodrigues
dc.subject.por.fl_str_mv Candida
Biofilme
Suscetibilidade
Antifúngicos
Infecções
topic Candida
Biofilme
Suscetibilidade
Antifúngicos
Infecções
Biofilm
Susceptibility
Antifungals
Infections
BIOLOGIA E FISIOLOGIA DOS MICROORGANISMOS::MICOLOGIA
dc.subject.eng.fl_str_mv Biofilm
Susceptibility
Antifungals
Infections
dc.subject.cnpq.fl_str_mv BIOLOGIA E FISIOLOGIA DOS MICROORGANISMOS::MICOLOGIA
description C. albicans infections may be favored by tissue damages such insertion of medical devices that function as a substrate for fungus growth. This growth can occur in the form of three-dimensional structures formed by yeast and hyphae, coated with a matrix of extracellular polymeric material, forming a biofilm. This agregated of cells is characterized by having greater resistance to host defenses and antifungals, which makes hard treating these infections. This study aimed to determine the kinetics of biofilm formation by isolates of Candida sp. and to evaluate the antifungal susceptibility of plancktonic and sessile cells. We evaluated 10 isolates of C. albicans from different materials, isolated and identified by standardized methods. The minimum inhibitory concentrations (MICs) and minimal fungicidal concentrations (MFCs) of amphotericin B, fluconazole, voriconazole and itraconazole to plancktonic cells were performed according to protocol M27-A3 of Clinical Laboratory Standard Institute (CLSI). Verification of kinetics of biofilms’ formation was assessed at 24 h, 48 h, 72 h and 96 h of incubation at 37 ° C and in vitro susceptibility of sessile cells, was evaluated using the MTT (3 - [4,5-Dimethyl-2-Thiazy] -2,5- Diphenyl-2H-Tetrazolium bromide) assay. The analysis of kinetics at 24 h showed that all isolates produced biofilm and the highest metabolic activity occurred within 48h of growth. Significant differences, on the metabolic activity of biofilms were observed on strains U02, M02 and V60, which produced the highest amount of biofilm with 48 h of growth. MIC values obtained for plancktonic cells ranged from 1 to 8 mg/mL for fluconazole; between 0.0625 to 1 mg/mL for itraconazole; between 0.0312 to 0.25 g/mL for voriconazole; and between 0.25 to 2 mg/mL for amphotericin B. The MFC was twice the MIC for all isolates. For the values obtained for fluconazole the MICS50 ranged from 8 mg/mL to > 1024 mg/mL and MICS80 ≥ 1024 mg/mL. MICS50 for itraconazole ranged from 4 mg/mL to > 16 g/mL and MICS80 ≥ 16 g/mL. The MICS50 for voriconazole showed values between 4 mg/mL and > 16 g/mL and MICS80 ≥ 16 g/mL. Amphotericin B had MICS50 values of 0.25 mg / mL to > 16 g/mL and values MICS80 of 2 mg/mL to > 16 g/mL. We could demonstrate an increased resistance of biofilms to major antifungal drugs by evaluating the susceptibility of C. albicans isolates as plancktonic and sessile cells. These results show the need of new therapeutic or preventive strategies to reduce the mortality rate of patients with infections associated with Candida biofilms.
publishDate 2011
dc.date.issued.fl_str_mv 2011-05-16
dc.date.accessioned.fl_str_mv 2019-01-22T10:53:29Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/masterThesis
format masterThesis
status_str publishedVersion
dc.identifier.citation.fl_str_mv PEREIRA, Núbia Pontes. Atividade in vitro de antifúngicos em biofilmes de Candida. 2011. 67 f. Dissertação (Mestrado em Medicina Tropical e Saúde Publica) - Universidade Federal de Goiás, Goiânia, 2011.
dc.identifier.uri.fl_str_mv http://repositorio.bc.ufg.br/tede/handle/tede/9240
dc.identifier.dark.fl_str_mv ark:/38995/0013000002h3s
identifier_str_mv PEREIRA, Núbia Pontes. Atividade in vitro de antifúngicos em biofilmes de Candida. 2011. 67 f. Dissertação (Mestrado em Medicina Tropical e Saúde Publica) - Universidade Federal de Goiás, Goiânia, 2011.
ark:/38995/0013000002h3s
url http://repositorio.bc.ufg.br/tede/handle/tede/9240
dc.language.iso.fl_str_mv por
language por
dc.relation.program.fl_str_mv 6085308344741430434
dc.relation.confidence.fl_str_mv 600
600
600
dc.relation.department.fl_str_mv -7769011444564556288
dc.relation.cnpq.fl_str_mv 1258845414555579790
dc.rights.driver.fl_str_mv http://creativecommons.org/licenses/by-nc-nd/4.0/
info:eu-repo/semantics/openAccess
rights_invalid_str_mv http://creativecommons.org/licenses/by-nc-nd/4.0/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Universidade Federal de Goiás
dc.publisher.program.fl_str_mv Programa de Pós-graduação em Medicina Tropical e Saúde Publica (IPTSP)
dc.publisher.initials.fl_str_mv UFG
dc.publisher.country.fl_str_mv Brasil
dc.publisher.department.fl_str_mv Instituto de Patologia Tropical e Saúde Pública - IPTSP (RG)
publisher.none.fl_str_mv Universidade Federal de Goiás
dc.source.none.fl_str_mv reponame:Repositório Institucional da UFG
instname:Universidade Federal de Goiás (UFG)
instacron:UFG
instname_str Universidade Federal de Goiás (UFG)
instacron_str UFG
institution UFG
reponame_str Repositório Institucional da UFG
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repository.name.fl_str_mv Repositório Institucional da UFG - Universidade Federal de Goiás (UFG)
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