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Master's Dissertation
DOI
https://doi.org/10.11606/D.76.2023.tde-07032023-101350
Document
Author
Full name
Maria Júlia de Arruda Mazzotti Marques
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Carlos, 2023
Supervisor
Committee
Kurachi, Cristina (President)
Kress, Marcia Regina von Zeska
Oliveira, Kleber Thiago de
Title in Portuguese
Estudo in vitro da inativação fotodinâmica do Rhizopus oryzae
Keywords in Portuguese
Rhizopus oryzae
Controle microbiano
Inativação fotodinâmica
Mucormicose
Abstract in Portuguese
Durante a pandemia da COVID-19, surgiram várias complicações em pacientes infectados, sendo uma delas a mucormicose, que é uma doença fúngica extremamente agressiva com uma elevada taxa de mortalidade, especialmente em pessoas com sistema imune comprometidos. A maioria dos casos de mucormicose é causada pelo fungo Rhizopus oryzae, também conhecido como fungo negro, com 90% dos casos afetando o sítio rinocerebral. Os tratamentos utilizados são baseados em doses elevadas de anfotericina B e posaconazol, associadas a ressecções cirúrgicas quando possível. Contudo, mesmo com um tratamento antifúngico agressivo, a taxa de mortalidade atribuível estimada é elevada. Na ausência de desbridamento cirúrgico do tecido infectado, o tratamento antifúngico por si só não é curativo. Por isso há necessidade de desenvolvimento de tratamentos adjuvantes. A Terapia Fotodinâmica antimicrobiana (TFDa) pode ser uma opção terapêutica auxiliar para a mucormicose. Devido à falta de relatos na literatura sobre a morfologia e a inativação fotodinâmica de R. oryzae, realizamos uma caracterização desse fungo utilizando Microscopia Confocal e Microscopia Eletrônica de transmissão e investigamos diferentes protocolos utilizando o Photodithazine® (PDZ) como um fotossensibilizador. Foi estudada resposta sobre a taxa de crescimento do fungo sob parâmetros de tratamento distintos como concentração e tempo de incubação do fotossensibilizador e associação com surfactante Dodecil Sulfato de Sódio (SDS). Para as hifas, tanto na fase clara quanto escura, nos protocolos utilizando somente PDZ observamos uma resposta fotodinâmica ineficaz, ao adicionar SDS 0,01% notamos uma melhora na resposta e com a combinação do SDS 0,05% e PDZ obtivemos uma taxa de inibição de 98% do crescimento para 2 sessões de TFDa na fase clara e 72% de inibição no protocolo de 1 sessão para a fase escura. No estudo em conídio conseguimos observar a redução de 1,7 log10 do crescimento do fungo. Neste presente trabalho, a TFDa mostrou potencial antimicrobiano no estudo in vitro do R. oryzae.
Title in English
In vitro study of photodynamic inactivation of Rhizopus oryzae
Keywords in English
Rhizopus oryzae
Microbial control
Mucormycosis
Photodynamic inactivation
Abstract in English
During the COVID-19 pandemic, several complications arose in infected patients, one of them was mucormycosis, which is an extremely aggressive fungal disease with a high mortality rate, especially in people with compromised immune system. The majority of the mucormycosis cases are caused by the fungus Rhizopus oryzae, also known as black fungus, with 90% of cases affecting the rhinocerebral region. The treatment used are based on high doses of amphotericin B and posaconazole, associated with surgical resection whenever possible. However, even with an aggressive antifungal treatment, the estimated attributable mortality rate is high. In the absence of surgical debridement of the infected tissue, antifungal treatment alone is not curative. Therefore, there is a need for development of alternative auxiliary treatments. Antimicrobial Photodynamic Therapy (aPDT) may be an auxiliary therapeutic option for mucormycosis. The purpose of this study is to evaluate the efficiency of photodynamic inactivation in controlling the growth of R. oryzae. Due to the lack of reports in the literature on the morphology and photodynamic inactivation of R. oryzae, we performed a characterization of this fungus using Confocal Microscopy and Transmission Electron Microscopy and investigated different protocols using Photodithazine® (PDZ) as a photosensitizer. Response on fungal growth rate under different treatment parameters such as photosensitizer concentration and incubation time, and association with Sodium Dodecyl Sulfate (SDS) surfactant was studied. For hyphae, both in the light and dark phases, in the protocols using only PDZ we observed an ineffective photodynamic response, when adding SDS 0.01% we noticed an improvement in the response and with the combination of SDS 0.05% and PDZ we obtained a 98% growth inhibition rate for 2 sessions of aPDT in the light phase and 72% inhibition in the 1 session protocol for the black phase. In the conidium study we were able to observe 1.7 log10 reduction in the fungus growth. In this present work, TFDa showed great potential in the in vitro study of R. oryzae.
 
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Publishing Date
2023-03-07
 
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