Sporothrix schenckii complex biofilms: in vitro formation and susceptibility
Autor(a) principal: | |
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Data de Publicação: | 2016 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Biblioteca Digital de Teses e Dissertações da UFC |
Texto Completo: | http://www.teses.ufc.br/tde_busca/arquivo.php?codArquivo=18159 |
Resumo: | Sporotrichosis is a cosmopolitan subcutaneous mycosis, found mainly in tropical and subtropical areas of Latin America, caused by species of Sporothrix schenckii complex. The aim of this study was to demonstrate the capacity of Sporothrix schenckii complex to produce in vitro biofilms and to determine the profile of these biofilms against to classical antifungal drugs. Four species from Sporothrix schenckii complex were used: S. brasiliensis (n=10), S. schenckii sensu stricto (n=2), S. globosa (n=2) and S. mexicana (n=4). The formation of biofilms was performed by transferring inoculum of 2 x 105 conidia/mL for microdilution plates, which were incubated at 25 ÂC for 5 days. After each day of incubation, XTT and Crystal Violet assays were made to determine the metabolic activity and biomass of biofilm, respectively. For susceptibility tests, three different concentrations (planktonic MIC, 10 x MIC and 50 x MIC) of antifungals amphotericin B (AMB), caspofungin (CAS), ketoconazole (KTC), voriconazole (VRC) and fluconazole (FLC) were transferred to biofilm wells, which were incubated for 72 hours at 25 ÂC. The biofilms profile then antifungal drugs presence was determined by the XTT and crystal violet assays. Biofilms were produced in Thermanoxâ coverslips for viewing by optical microscopy with congo-red staining, scanning electron microscopy and confocal microscopy. It was found that all species from S. schenckii complex are strong biofilm-forming, with no difference formation between them. Absorbance of biofilm after drug exposure showed that biomass and metabolic activity was significantly reduced (p<0.05), mainly in S. brasiliensis. Antifungal drugs tested showed metabolic activity and biomass reduction, especially at concentrations 50 times higher than the planktonic MICs, demonstrating the protective increase in sessile form when compared for planktonic form. |
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info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisSporothrix schenckii complex biofilms: in vitro formation and susceptibilityBiofilmes do complexo Sporothrix schenckii: formaÃÃo e sensibilidade in vitro2016-11-17Raimunda SÃmia Nogueira Brilhante70399573372http://lattes.cnpq.br/4766125121792218FlÃvia Almeida Santos48438421334http://buscatextual.cnpq.br/buscatextual/visualizacv.jsp?id=K4791154J9Adriana de Queiroz Pinheiro3815265738701130182312Felipe Rodrigues MagalhÃes de AguiarUniversidade Federal do CearÃPrograma de PÃs-GraduaÃÃo em Microbiologia MÃdicaUFCBRSensibilidadeAntifÃngicoMICROBIOLOGIA MEDICASporotrichosis is a cosmopolitan subcutaneous mycosis, found mainly in tropical and subtropical areas of Latin America, caused by species of Sporothrix schenckii complex. The aim of this study was to demonstrate the capacity of Sporothrix schenckii complex to produce in vitro biofilms and to determine the profile of these biofilms against to classical antifungal drugs. Four species from Sporothrix schenckii complex were used: S. brasiliensis (n=10), S. schenckii sensu stricto (n=2), S. globosa (n=2) and S. mexicana (n=4). The formation of biofilms was performed by transferring inoculum of 2 x 105 conidia/mL for microdilution plates, which were incubated at 25 ÂC for 5 days. After each day of incubation, XTT and Crystal Violet assays were made to determine the metabolic activity and biomass of biofilm, respectively. For susceptibility tests, three different concentrations (planktonic MIC, 10 x MIC and 50 x MIC) of antifungals amphotericin B (AMB), caspofungin (CAS), ketoconazole (KTC), voriconazole (VRC) and fluconazole (FLC) were transferred to biofilm wells, which were incubated for 72 hours at 25 ÂC. The biofilms profile then antifungal drugs presence was determined by the XTT and crystal violet assays. Biofilms were produced in Thermanoxâ coverslips for viewing by optical microscopy with congo-red staining, scanning electron microscopy and confocal microscopy. It was found that all species from S. schenckii complex are strong biofilm-forming, with no difference formation between them. Absorbance of biofilm after drug exposure showed that biomass and metabolic activity was significantly reduced (p<0.05), mainly in S. brasiliensis. Antifungal drugs tested showed metabolic activity and biomass reduction, especially at concentrations 50 times higher than the planktonic MICs, demonstrating the protective increase in sessile form when compared for planktonic form.A esporotricose à uma micose subcutÃnea cosmopolita, encontrada, principalmente, em Ãreas tropicais e subtropicais da AmÃrica Latina, sendo causada por espÃcies do complexo Sporothrix schenckii. O objetivo deste estudo foi demonstrar a capacidade de fungos do complexo S. schenckii de produzir biofilmes in vitro e determinar o perfil desses biofilmes frente Ãs drogas antifÃngicas clÃssicas. Foram utilizadas 4 espÃcies do complexo Sporothrix schenckii: S. brasiliensis (n=10), S. schenckii sensu stricto (n=2), S. globosa (n=2) e S. mexicana (n=4). A formaÃÃo dos biofilmes foi feita atravÃs da transferÃncia de inÃculos de 2 x 105 conÃdios/mL para placas de microdiluiÃÃo, que foram incubadas a 25 ÂC por 5 dias. ApÃs cada dia de incubaÃÃo, realizaram-se ensaios de XTT e Cristal violeta, para se determinar a atividade metabÃlica e biomassa dos biofilmes, respectivamente. Para os ensaios de sensibilidade, trÃs concentraÃÃes diferentes (MIC planctÃnico, 10 x MIC e 50 x MIC) dos antifÃngicos anfotericina B (AMB), caspofungina (CAS), cetoconazol (KTC), voriconazol (VRC) e fluconazol (FLC) foram transferidas para os poÃos com biofilmes, que foram incubados por 72 horas a 25 ÂC. O perfil dos biofilmes frente Ãs drogas antifÃngicas foi determinado atravÃs de ensaios de XTT e cristal violeta. Biofilmes foram produzidos em lamÃnulas de Thermanoxâ para visualizaÃÃo atravÃs de microscopia Ãptica com coloraÃÃo de vermelho-congo, microscopia eletrÃnica de varredura e microscopia confocal. Verificou-se que todas as espÃcies do complexo S. schenkii sÃo fortes formadoras de biofilme, nÃo havendo diferenÃa de formaÃÃo entre as espÃcies. A absorbÃncia dos biofilmes apÃs exposiÃÃo Ãs drogas demonstrou que a biomassa e a atividade metabÃlica foram reduzidas de forma significativa (p<0,05), principalmente, em S. brasiliensis. Os antifÃngicos testados mostraram reduÃÃo na atividade metabÃlica e biomassa, principalmente, em concentraÃÃes 50 vezes maiores que os MICs planctÃnicos, demonstrando o incremento protetivo que a forma sÃssil possui frente à forma planctÃnica.CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel SuperiorConselho Nacional de Desenvolvimento CientÃfico e TecnolÃgicohttp://www.teses.ufc.br/tde_busca/arquivo.php?codArquivo=18159application/pdfinfo:eu-repo/semantics/openAccessporreponame:Biblioteca Digital de Teses e Dissertações da UFCinstname:Universidade Federal do Cearáinstacron:UFC2019-01-21T11:31:18Zmail@mail.com - |
dc.title.en.fl_str_mv |
Sporothrix schenckii complex biofilms: in vitro formation and susceptibility |
dc.title.alternative.pt.fl_str_mv |
Biofilmes do complexo Sporothrix schenckii: formaÃÃo e sensibilidade in vitro |
title |
Sporothrix schenckii complex biofilms: in vitro formation and susceptibility |
spellingShingle |
Sporothrix schenckii complex biofilms: in vitro formation and susceptibility Felipe Rodrigues MagalhÃes de Aguiar Sensibilidade AntifÃngico MICROBIOLOGIA MEDICA |
title_short |
Sporothrix schenckii complex biofilms: in vitro formation and susceptibility |
title_full |
Sporothrix schenckii complex biofilms: in vitro formation and susceptibility |
title_fullStr |
Sporothrix schenckii complex biofilms: in vitro formation and susceptibility |
title_full_unstemmed |
Sporothrix schenckii complex biofilms: in vitro formation and susceptibility |
title_sort |
Sporothrix schenckii complex biofilms: in vitro formation and susceptibility |
author |
Felipe Rodrigues MagalhÃes de Aguiar |
author_facet |
Felipe Rodrigues MagalhÃes de Aguiar |
author_role |
author |
dc.contributor.advisor1.fl_str_mv |
Raimunda SÃmia Nogueira Brilhante |
dc.contributor.advisor1ID.fl_str_mv |
70399573372 |
dc.contributor.advisor1Lattes.fl_str_mv |
http://lattes.cnpq.br/4766125121792218 |
dc.contributor.referee1.fl_str_mv |
FlÃvia Almeida Santos |
dc.contributor.referee1ID.fl_str_mv |
48438421334 |
dc.contributor.referee1Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.jsp?id=K4791154J9 |
dc.contributor.referee2.fl_str_mv |
Adriana de Queiroz Pinheiro |
dc.contributor.referee2ID.fl_str_mv |
38152657387 |
dc.contributor.authorID.fl_str_mv |
01130182312 |
dc.contributor.author.fl_str_mv |
Felipe Rodrigues MagalhÃes de Aguiar |
contributor_str_mv |
Raimunda SÃmia Nogueira Brilhante FlÃvia Almeida Santos Adriana de Queiroz Pinheiro |
dc.subject.por.fl_str_mv |
Sensibilidade AntifÃngico |
topic |
Sensibilidade AntifÃngico MICROBIOLOGIA MEDICA |
dc.subject.cnpq.fl_str_mv |
MICROBIOLOGIA MEDICA |
dc.description.sponsorship.fl_txt_mv |
CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior Conselho Nacional de Desenvolvimento CientÃfico e TecnolÃgico |
dc.description.abstract.por.fl_txt_mv |
Sporotrichosis is a cosmopolitan subcutaneous mycosis, found mainly in tropical and subtropical areas of Latin America, caused by species of Sporothrix schenckii complex. The aim of this study was to demonstrate the capacity of Sporothrix schenckii complex to produce in vitro biofilms and to determine the profile of these biofilms against to classical antifungal drugs. Four species from Sporothrix schenckii complex were used: S. brasiliensis (n=10), S. schenckii sensu stricto (n=2), S. globosa (n=2) and S. mexicana (n=4). The formation of biofilms was performed by transferring inoculum of 2 x 105 conidia/mL for microdilution plates, which were incubated at 25 ÂC for 5 days. After each day of incubation, XTT and Crystal Violet assays were made to determine the metabolic activity and biomass of biofilm, respectively. For susceptibility tests, three different concentrations (planktonic MIC, 10 x MIC and 50 x MIC) of antifungals amphotericin B (AMB), caspofungin (CAS), ketoconazole (KTC), voriconazole (VRC) and fluconazole (FLC) were transferred to biofilm wells, which were incubated for 72 hours at 25 ÂC. The biofilms profile then antifungal drugs presence was determined by the XTT and crystal violet assays. Biofilms were produced in Thermanoxâ coverslips for viewing by optical microscopy with congo-red staining, scanning electron microscopy and confocal microscopy. It was found that all species from S. schenckii complex are strong biofilm-forming, with no difference formation between them. Absorbance of biofilm after drug exposure showed that biomass and metabolic activity was significantly reduced (p<0.05), mainly in S. brasiliensis. Antifungal drugs tested showed metabolic activity and biomass reduction, especially at concentrations 50 times higher than the planktonic MICs, demonstrating the protective increase in sessile form when compared for planktonic form. A esporotricose à uma micose subcutÃnea cosmopolita, encontrada, principalmente, em Ãreas tropicais e subtropicais da AmÃrica Latina, sendo causada por espÃcies do complexo Sporothrix schenckii. O objetivo deste estudo foi demonstrar a capacidade de fungos do complexo S. schenckii de produzir biofilmes in vitro e determinar o perfil desses biofilmes frente Ãs drogas antifÃngicas clÃssicas. Foram utilizadas 4 espÃcies do complexo Sporothrix schenckii: S. brasiliensis (n=10), S. schenckii sensu stricto (n=2), S. globosa (n=2) e S. mexicana (n=4). A formaÃÃo dos biofilmes foi feita atravÃs da transferÃncia de inÃculos de 2 x 105 conÃdios/mL para placas de microdiluiÃÃo, que foram incubadas a 25 ÂC por 5 dias. ApÃs cada dia de incubaÃÃo, realizaram-se ensaios de XTT e Cristal violeta, para se determinar a atividade metabÃlica e biomassa dos biofilmes, respectivamente. Para os ensaios de sensibilidade, trÃs concentraÃÃes diferentes (MIC planctÃnico, 10 x MIC e 50 x MIC) dos antifÃngicos anfotericina B (AMB), caspofungina (CAS), cetoconazol (KTC), voriconazol (VRC) e fluconazol (FLC) foram transferidas para os poÃos com biofilmes, que foram incubados por 72 horas a 25 ÂC. O perfil dos biofilmes frente Ãs drogas antifÃngicas foi determinado atravÃs de ensaios de XTT e cristal violeta. Biofilmes foram produzidos em lamÃnulas de Thermanoxâ para visualizaÃÃo atravÃs de microscopia Ãptica com coloraÃÃo de vermelho-congo, microscopia eletrÃnica de varredura e microscopia confocal. Verificou-se que todas as espÃcies do complexo S. schenkii sÃo fortes formadoras de biofilme, nÃo havendo diferenÃa de formaÃÃo entre as espÃcies. A absorbÃncia dos biofilmes apÃs exposiÃÃo Ãs drogas demonstrou que a biomassa e a atividade metabÃlica foram reduzidas de forma significativa (p<0,05), principalmente, em S. brasiliensis. Os antifÃngicos testados mostraram reduÃÃo na atividade metabÃlica e biomassa, principalmente, em concentraÃÃes 50 vezes maiores que os MICs planctÃnicos, demonstrando o incremento protetivo que a forma sÃssil possui frente à forma planctÃnica. |
description |
Sporotrichosis is a cosmopolitan subcutaneous mycosis, found mainly in tropical and subtropical areas of Latin America, caused by species of Sporothrix schenckii complex. The aim of this study was to demonstrate the capacity of Sporothrix schenckii complex to produce in vitro biofilms and to determine the profile of these biofilms against to classical antifungal drugs. Four species from Sporothrix schenckii complex were used: S. brasiliensis (n=10), S. schenckii sensu stricto (n=2), S. globosa (n=2) and S. mexicana (n=4). The formation of biofilms was performed by transferring inoculum of 2 x 105 conidia/mL for microdilution plates, which were incubated at 25 ÂC for 5 days. After each day of incubation, XTT and Crystal Violet assays were made to determine the metabolic activity and biomass of biofilm, respectively. For susceptibility tests, three different concentrations (planktonic MIC, 10 x MIC and 50 x MIC) of antifungals amphotericin B (AMB), caspofungin (CAS), ketoconazole (KTC), voriconazole (VRC) and fluconazole (FLC) were transferred to biofilm wells, which were incubated for 72 hours at 25 ÂC. The biofilms profile then antifungal drugs presence was determined by the XTT and crystal violet assays. Biofilms were produced in Thermanoxâ coverslips for viewing by optical microscopy with congo-red staining, scanning electron microscopy and confocal microscopy. It was found that all species from S. schenckii complex are strong biofilm-forming, with no difference formation between them. Absorbance of biofilm after drug exposure showed that biomass and metabolic activity was significantly reduced (p<0.05), mainly in S. brasiliensis. Antifungal drugs tested showed metabolic activity and biomass reduction, especially at concentrations 50 times higher than the planktonic MICs, demonstrating the protective increase in sessile form when compared for planktonic form. |
publishDate |
2016 |
dc.date.issued.fl_str_mv |
2016-11-17 |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/masterThesis |
status_str |
publishedVersion |
format |
masterThesis |
dc.identifier.uri.fl_str_mv |
http://www.teses.ufc.br/tde_busca/arquivo.php?codArquivo=18159 |
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http://www.teses.ufc.br/tde_busca/arquivo.php?codArquivo=18159 |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
Universidade Federal do Cearà |
dc.publisher.program.fl_str_mv |
Programa de PÃs-GraduaÃÃo em Microbiologia MÃdica |
dc.publisher.initials.fl_str_mv |
UFC |
dc.publisher.country.fl_str_mv |
BR |
publisher.none.fl_str_mv |
Universidade Federal do Cearà |
dc.source.none.fl_str_mv |
reponame:Biblioteca Digital de Teses e Dissertações da UFC instname:Universidade Federal do Ceará instacron:UFC |
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Biblioteca Digital de Teses e Dissertações da UFC |
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Biblioteca Digital de Teses e Dissertações da UFC |
instname_str |
Universidade Federal do Ceará |
instacron_str |
UFC |
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UFC |
repository.name.fl_str_mv |
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repository.mail.fl_str_mv |
mail@mail.com |
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1643295229139222528 |