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Doctoral Thesis
DOI
https://doi.org/10.11606/T.60.2021.tde-29092021-053538
Document
Author
Full name
Mario Henrique Paziani
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
Ribeirão Preto, 2021
Supervisor
Committee
Kress, Marcia Regina von Zeska (President)
Martinez, Roberto
Cabral, Hamilton
Lima, Nelson Manuel Viana da Silva
Muñoz, María Antonieta Ruiz
Santos, Cledir Rodrigues
Title in Portuguese
Perfil fenotípico e genotípico de Fusarium spp. provenientes de episódios clínicos de fusarioses no Estado de São Paulo
Keywords in Portuguese
Biofilme
Fusariose
Fusarium sp.
Galleria mellonella
Identificação molecular
MALDI-TOF MS
Terapia fotodinâmica antimicrobiana
Virulência
Abstract in Portuguese
O gênero Fusarium apresenta espécies de fungos filamentosos do filo Ascomycota e são majoritariamente isolados em regiões tropicais e subtropicais de uma grande variedade de substratos, como solo e plantas. Em humanos, a fusariose subcutânea aumentou em níveis alarmantes e pode progredir à infecção invasiva, devido ao crescente número de pacientes imunocomprometidos. Os potenciais patógenos humanos do gênero Fusarium são membros dos complexos de espécies FOSC, FSSC, FFSC e FDSC. O objetivo do presente estudo foi identificar e caracterizar 108 isolados clínicos de Fusarium spp. provenientes de infecções em pacientes do Estado de São Paulo, Brasil. Estes foram identificados pelo método molecular de sequenciamento das regiões ITS do rDNA e do gene EF1α, e pelo método fenotípico de MALDI-TOF MS. Os isolados clínicos foram avaliados quanto a capacidade de formar biofilme, susceptibilidade aos antifúngicos e à terapia fotodinâmica antimicrobiana (TFDA) e virulência no modelo alternativo G. mellonella. Entre os isolados clínicos do FSSC estão F. keratoplasticum (n=58; 59,7%), F. solani (n=28; 28,8%), F. falciforme (n=9; 9,2%) e F. petroliphilum (n=2; 2,0%). No FOSC, todos os isolados clínicos (n=9; 100%) são F. oxysporum, enquanto que para FFSC (n=1; 100%) e FDSC (n=1; 100%), os isolados clínicos são F. proliferatum e F. delphinoides, respectivamente. MALDI-TOF MS identificou e agrupou a maioria dos isolados clínicos nos complexos de espécies de Fusarium. Os isolados clínicos de FSSC apresentaram maior capacidade de produção de biofilme comparando com FOSC. A maior parte dos isolados clínicos de Fusarium spp. apresentam altas CIMs in vitro de itraconazol. Além disso, a TFDA com fotossensibilizadores fenotiazínicos foi eficaz in vitro e in vivo contra todas as espécies de Fusarium. FSSC apresenta maior virulência em G. mellonella, em comparação com os outros complexos de espécie. Nossos estudos expandem a compreensão do perfil fenotípico e genotípico de Fusarium spp. no Estado de São Paulo, Brasil.
Title in English
Phenotypic and genotypic profile of Fusarium spp. from clinical episodes of fusariosis in the State of São Paulo
Keywords in English
Antimicrobial photodynamic therapy
Biofilm
Fusariosis
Fusarium sp.
MALDI-TOF MS
Molecular identification
Virulence
Abstract in English
Fusarium genus is composed of filamentous fungi of the phylum Ascomycota and are mostly isolated in tropical regions, in a wide variety of substrates, such as soil and plants. In humans, subcutaneous fusariosis has increased to alarming levels and can progress to invasive infection, due to the increasing number of immunocompromised patients. The potential human pathogens among Fusarium spp. are members of FOSC, FSSC, FFSC, and FDSC. The present study aimed to identify and characterize 108 clinical isolates of Fusarium spp. from infections in patients from the State of São Paulo, Brazil. The clinical isolates were identified by sequencing the ITS regions of the rDNA and the EF1α gene and with the phenotypic method MALDI-TOF MS. The clinical isolates were evaluated for their ability to form a biofilm, in vitro antifungal susceptibility to commercial antifungal drugs, in vitro and in vivo susceptibility to antimicrobial photodynamic therapy (APDT), and virulence analyzes in the alternative model Galleria mellonella. Among the clinical isolates of the FSSC are F. keratoplasticum (n=58; 59.7%), F. solani (n=28; 28.8%), F. falciforme (n=9; 9.2%), and F. petroliphilum (n=2; 2.0%). In FOSC, all clinical isolates (n=9; 100%) are F. oxysporum, while for FFSC (n=1; 100%) and FDSC (n=1; 100%), clinical isolates are F. proliferatum and F. delphinoides, respectively. MALDI-TOF MS identified and grouped the Fusarium clinical isolates in the species complexes. The clinical isolates of FSSC showed greater biofilm production capacity compared to FOSC. Most of the Fusarium spp. have shown high MIC of itraconazole and FSSC has a higher virulence in G. mellonella, compared to other species complexes. In addition, TFDA with phenothiazine photosensitizers has been effective in vitro and in vivo against all Fusarium species. Our studies expand the understanding of the phenotypic and genotypic profile of Fusarium spp. in the State of Sao Paulo, Brazil.
 
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Release Date
2023-07-27
Publishing Date
2021-09-29
 
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