Cepas do complexo Candida parapsilosis de origem animal : classificação taxonômica, sensibilidade antifúngica e atributos de virulência in vitro

Detalhes bibliográficos
Autor(a) principal: Rodrigues, Terezinha de Jesus Santos
Data de Publicação: 2013
Tipo de documento: Dissertação
Idioma: por
Título da fonte: Repositório Institucional da Universidade Federal do Ceará (UFC)
dARK ID: ark:/83112/001300000cxmg
Texto Completo: http://www.repositorio.ufc.br/handle/riufc/7574
Resumo: In the past decades, important changes in the epidemiology of fungal infections have resulted in the frequent isolation of yeasts of the genus Candida. Yeasts of the Candida parapsilosis species complex, for example, have been pointed out as infectious agents and components of the microbiota of animals. Thus, the work aimed at identifying molecularly the strains of the C. parapsilosis species complex recovered from veterinary sources and maintained at the Specialized Medical Mycology Center, as well as evaluating their in vitro antifungal susceptibility profile and attributes of virulence. For such, 28 strains of the C. parapsilosis species complex, recovered from dogs, psittacines, raptors and prawn were assessed. Initially, the strains were phenotypically identified, based on their morphological and biochemical characteristics, which confirmed their identification as C. parapsilosis lato sensu. The molecular identification of the strains was then carried out through PCR-REA. In order to analyze the in vitro antifungal susceptibility, the broth microdilution assay was performed, according to the document M27-A3 of the Clinical Laboratory Standards Institute (CLSI) 2008, using amphotericin B, itraconazole, voriconazole, fluconazole and caspofungin. Concerning the virulence attributes, the ability of producing phospholipase was evaluated on egg yolk agar, while protease production was assessed on bovine serum albumin agar, and biofilm formation was tested in 96-well polystyrene microplates. The genotypical analysis identified 13 C. parapsilosis stricto sensu, ten C. orthopsilosis and five C. metapsilosis. The minimum inhibitory concentrations varied from 0.125 to 1 μg/mL foramphotericin B, from 0.03125 to 0.5 μg/mL, for itraconazole, from 0.03125 to 0,25 μg/mL for voriconazole, from 0.5 to 16 μg/mL for fluconazol and from 0.0625 to 2 μg/mL for caspofungin. Fluconazole resistance was observed in three strains of C. parapsilosis stricto sensu and two of C. metapsilosis, while one strain of C. parapsilosis stricto sensu and five C. orthopsilosis presented high MIC values for caspofungin (2 μg/mL). As for the virulence attributes, none of the tested strains produced phospholipases, while 23/28 presented proteolytic activity, and all of the strains produced biofilm, with one weak producer, 20 moderate producers and seven strong producers. These data show that the in vitro antifungal susceptibility and production of virulence attributes vary among species of the C. parapsilosis species complex, which can lead to differences in pathogenicity and therapeutic response.
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spelling Cepas do complexo Candida parapsilosis de origem animal : classificação taxonômica, sensibilidade antifúngica e atributos de virulência in vitroCandida parapsilosis complex from animals and its antifungal susceptibility and virulence attributesVirulênciaCandidaAnimaisIn the past decades, important changes in the epidemiology of fungal infections have resulted in the frequent isolation of yeasts of the genus Candida. Yeasts of the Candida parapsilosis species complex, for example, have been pointed out as infectious agents and components of the microbiota of animals. Thus, the work aimed at identifying molecularly the strains of the C. parapsilosis species complex recovered from veterinary sources and maintained at the Specialized Medical Mycology Center, as well as evaluating their in vitro antifungal susceptibility profile and attributes of virulence. For such, 28 strains of the C. parapsilosis species complex, recovered from dogs, psittacines, raptors and prawn were assessed. Initially, the strains were phenotypically identified, based on their morphological and biochemical characteristics, which confirmed their identification as C. parapsilosis lato sensu. The molecular identification of the strains was then carried out through PCR-REA. In order to analyze the in vitro antifungal susceptibility, the broth microdilution assay was performed, according to the document M27-A3 of the Clinical Laboratory Standards Institute (CLSI) 2008, using amphotericin B, itraconazole, voriconazole, fluconazole and caspofungin. Concerning the virulence attributes, the ability of producing phospholipase was evaluated on egg yolk agar, while protease production was assessed on bovine serum albumin agar, and biofilm formation was tested in 96-well polystyrene microplates. The genotypical analysis identified 13 C. parapsilosis stricto sensu, ten C. orthopsilosis and five C. metapsilosis. The minimum inhibitory concentrations varied from 0.125 to 1 μg/mL foramphotericin B, from 0.03125 to 0.5 μg/mL, for itraconazole, from 0.03125 to 0,25 μg/mL for voriconazole, from 0.5 to 16 μg/mL for fluconazol and from 0.0625 to 2 μg/mL for caspofungin. Fluconazole resistance was observed in three strains of C. parapsilosis stricto sensu and two of C. metapsilosis, while one strain of C. parapsilosis stricto sensu and five C. orthopsilosis presented high MIC values for caspofungin (2 μg/mL). As for the virulence attributes, none of the tested strains produced phospholipases, while 23/28 presented proteolytic activity, and all of the strains produced biofilm, with one weak producer, 20 moderate producers and seven strong producers. These data show that the in vitro antifungal susceptibility and production of virulence attributes vary among species of the C. parapsilosis species complex, which can lead to differences in pathogenicity and therapeutic response.Mudanças importantes na epidemiologia das infecções fúngicas nas últimas décadas têm resultado no isolamento frequente de leveduras do gênero Candida. As leveduras do complexo Candida parapsilosis, por exemplo, têm sido apontadas tanto como agentes de infecções como componentes da microbiota de animais. Diante disso, os objetivos deste trabalho foram realizar a identificação molecular de cepas do complexo C. parapsilosis, isoladas de fontes veterinárias e mantidas no Centro Especializado em Micologia Médica; assim como avaliar seus atributos de virulência e perfil de sensibilidade a antifúngicos, in vitro. Para tanto, foram utilizados 28 cepas do complexo C. parapsilosis obtidos de cães, psitacídeos, rapinantes e camarão. Inicialmente foi realizada a fenotipagem das cepas, com base na análise de suas características morfológicas e bioquímicas, que as ratificou como C. parapsilosis (lato sensu). A identificação molecular das espécies foi realizada por PCR-REA. A fim de analisar o perfil de sensibilidade das cepas, empregou-se o teste de microdiluição em caldo com anfotericina B, itraconazol, fluconazol, voriconazol e caspofungina, segundo metodologia padronizada pelo M27-A3. CLSI (2008). No tocante aos atributos de virulência, a capacidade da produção de fosfolipases foi avaliada pelo método de cultivo em ágar gema de ovo, a produção de proteases foi analisada através de cultivo em ágar albumina bovina e a formação de biofilme foi em microplaca de poliestireno com 96 poços. A análise genotípica evidenciou 13 C. parapsilosis (stricto sensu), 10 C. orthopsilosis e 05 C. metapsilosis. As concentrações inibitórias mínimas (MICs) variaram de 0,125 a 1 μg/mL para anfotericina B, de 0,5 a 16 μg/mL para fluconazol, de 0,03125 a 0,5 μg/mL para itraconazol, de 0,03125 a 0,25 μg/mL para voriconazol e de 0,0625 a 2 μg/mL para caspofungina. Foi observada resistência em 03 cepas de C. parapsilosis (stricto sensu) ao fluconazol e 01 cepa apresentou MIC elevado (2 μg/mL) para caspofungina. No que tange aos isolados de C. orthopsilosis, notou-se que 05 isolados apresentaram MICs elevados (2 μg/mL) para a caspofungina. Enquanto que, 02 isolados de C. metapsilosis revelaram-se resistentes ao fluconazol. Quanto à virulência, todas as cepas foram capazes de formar biofilmes, sendo, 20, 7 e 01, classificadas como produtoras moderadas, fortes e fracas respectivamente. Observou-se, ainda que 23/28 isolados apresentaram atividade proteolítica. Por outro lado, nenhuma foi capaz de produzir fosfolipases. Estes dados sinalizam que padrões de virulência, patogenicidade sensibilidade antifúngica, in vitro, podem variar entre as espécies do complexo C. parapsilosis.Rocha, Marcos Fábio GadelhaRodrigues, Terezinha de Jesus Santos2014-03-07T15:27:27Z2014-03-07T15:27:27Z2013info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfRODRIGUES, T. J. S. Cepas do complexo Candida parapsilosis de origem animal : classificação taxonômica, sensibilidade antifúngica e atributos de virulência in vitro. 2013 .41 f. Dissertação (Mestrado em Microbiliologia Médica) - Universidade Federal do Ceará. Faculdade de Medicina, Fortaleza, 2013.http://www.repositorio.ufc.br/handle/riufc/7574ark:/83112/001300000cxmgporreponame:Repositório Institucional da Universidade Federal do Ceará (UFC)instname:Universidade Federal do Ceará (UFC)instacron:UFCinfo:eu-repo/semantics/openAccess2021-02-05T17:48:56Zoai:repositorio.ufc.br:riufc/7574Repositório InstitucionalPUBhttp://www.repositorio.ufc.br/ri-oai/requestbu@ufc.br || repositorio@ufc.bropendoar:2024-09-11T18:36:57.118301Repositório Institucional da Universidade Federal do Ceará (UFC) - Universidade Federal do Ceará (UFC)false
dc.title.none.fl_str_mv Cepas do complexo Candida parapsilosis de origem animal : classificação taxonômica, sensibilidade antifúngica e atributos de virulência in vitro
Candida parapsilosis complex from animals and its antifungal susceptibility and virulence attributes
title Cepas do complexo Candida parapsilosis de origem animal : classificação taxonômica, sensibilidade antifúngica e atributos de virulência in vitro
spellingShingle Cepas do complexo Candida parapsilosis de origem animal : classificação taxonômica, sensibilidade antifúngica e atributos de virulência in vitro
Rodrigues, Terezinha de Jesus Santos
Virulência
Candida
Animais
title_short Cepas do complexo Candida parapsilosis de origem animal : classificação taxonômica, sensibilidade antifúngica e atributos de virulência in vitro
title_full Cepas do complexo Candida parapsilosis de origem animal : classificação taxonômica, sensibilidade antifúngica e atributos de virulência in vitro
title_fullStr Cepas do complexo Candida parapsilosis de origem animal : classificação taxonômica, sensibilidade antifúngica e atributos de virulência in vitro
title_full_unstemmed Cepas do complexo Candida parapsilosis de origem animal : classificação taxonômica, sensibilidade antifúngica e atributos de virulência in vitro
title_sort Cepas do complexo Candida parapsilosis de origem animal : classificação taxonômica, sensibilidade antifúngica e atributos de virulência in vitro
author Rodrigues, Terezinha de Jesus Santos
author_facet Rodrigues, Terezinha de Jesus Santos
author_role author
dc.contributor.none.fl_str_mv Rocha, Marcos Fábio Gadelha
dc.contributor.author.fl_str_mv Rodrigues, Terezinha de Jesus Santos
dc.subject.por.fl_str_mv Virulência
Candida
Animais
topic Virulência
Candida
Animais
description In the past decades, important changes in the epidemiology of fungal infections have resulted in the frequent isolation of yeasts of the genus Candida. Yeasts of the Candida parapsilosis species complex, for example, have been pointed out as infectious agents and components of the microbiota of animals. Thus, the work aimed at identifying molecularly the strains of the C. parapsilosis species complex recovered from veterinary sources and maintained at the Specialized Medical Mycology Center, as well as evaluating their in vitro antifungal susceptibility profile and attributes of virulence. For such, 28 strains of the C. parapsilosis species complex, recovered from dogs, psittacines, raptors and prawn were assessed. Initially, the strains were phenotypically identified, based on their morphological and biochemical characteristics, which confirmed their identification as C. parapsilosis lato sensu. The molecular identification of the strains was then carried out through PCR-REA. In order to analyze the in vitro antifungal susceptibility, the broth microdilution assay was performed, according to the document M27-A3 of the Clinical Laboratory Standards Institute (CLSI) 2008, using amphotericin B, itraconazole, voriconazole, fluconazole and caspofungin. Concerning the virulence attributes, the ability of producing phospholipase was evaluated on egg yolk agar, while protease production was assessed on bovine serum albumin agar, and biofilm formation was tested in 96-well polystyrene microplates. The genotypical analysis identified 13 C. parapsilosis stricto sensu, ten C. orthopsilosis and five C. metapsilosis. The minimum inhibitory concentrations varied from 0.125 to 1 μg/mL foramphotericin B, from 0.03125 to 0.5 μg/mL, for itraconazole, from 0.03125 to 0,25 μg/mL for voriconazole, from 0.5 to 16 μg/mL for fluconazol and from 0.0625 to 2 μg/mL for caspofungin. Fluconazole resistance was observed in three strains of C. parapsilosis stricto sensu and two of C. metapsilosis, while one strain of C. parapsilosis stricto sensu and five C. orthopsilosis presented high MIC values for caspofungin (2 μg/mL). As for the virulence attributes, none of the tested strains produced phospholipases, while 23/28 presented proteolytic activity, and all of the strains produced biofilm, with one weak producer, 20 moderate producers and seven strong producers. These data show that the in vitro antifungal susceptibility and production of virulence attributes vary among species of the C. parapsilosis species complex, which can lead to differences in pathogenicity and therapeutic response.
publishDate 2013
dc.date.none.fl_str_mv 2013
2014-03-07T15:27:27Z
2014-03-07T15:27:27Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/masterThesis
format masterThesis
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dc.identifier.uri.fl_str_mv RODRIGUES, T. J. S. Cepas do complexo Candida parapsilosis de origem animal : classificação taxonômica, sensibilidade antifúngica e atributos de virulência in vitro. 2013 .41 f. Dissertação (Mestrado em Microbiliologia Médica) - Universidade Federal do Ceará. Faculdade de Medicina, Fortaleza, 2013.
http://www.repositorio.ufc.br/handle/riufc/7574
dc.identifier.dark.fl_str_mv ark:/83112/001300000cxmg
identifier_str_mv RODRIGUES, T. J. S. Cepas do complexo Candida parapsilosis de origem animal : classificação taxonômica, sensibilidade antifúngica e atributos de virulência in vitro. 2013 .41 f. Dissertação (Mestrado em Microbiliologia Médica) - Universidade Federal do Ceará. Faculdade de Medicina, Fortaleza, 2013.
ark:/83112/001300000cxmg
url http://www.repositorio.ufc.br/handle/riufc/7574
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