Aspectos microbiológicos e ambientais de candidemias em hospital terciário (HC/FMB/UNESP/Botucatu) localizado na região centro-sul do estado de São Paulo, Brasil
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Data
2018-02-23
Autores
Giacobino, Juliana
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Universidade Estadual Paulista (Unesp)
Resumo
Infecções fúngicas causadas por leveduras constituem um dos maiores problemas em pacientes hospitalizados em todo o mundo e têm se tornado uma importante causa de morbidade e mortalidade. Embora Candida albicans seja a espécie mais frequentemente isolada e sua forma de infecção ocorra geralmente por translocação endógena, tem-se observado um aumento das espécies não-albicans, com destaque para o complexo C. parapsilosis, cuja principal forma de infecção é exógena, provavelmente pelas mãos dos profissionais de saúde. Este trabalho propôs estudar os aspectos microbiológicos e ambientais das candidemias, com especial atenção para o complexo Candida parapsilosis, no hospital terciário HC/FMB/UNESP, Botucatu, utilizando-se de métodos moleculares, busca ativa dos agentes no ambiente hospitalar (ar, superfícies e mãos de profissionais de saúde), bem como determinar os fatores de virulência e susceptibilidade aos antifúngicos destas espécies e associação com o desfecho clínico. Os isolados clínicos (obtidos de hemoculturas, período 2007-2015) e ambientais (período 2014-2015) de C. parapsilosis sensu lato foram identificados pelo meio CHROMagar Candida, Vitek-2, sequenciamento do rDNA e perfis dos inteins VMA e ThrRS. Fatores de virulência (produção de proteinase, fosfolipase e biofilme) e perfis de susceptibilidade aos antifúngicos anfotericina B, fluconazol, voriconazol, caspofungina e micafungina foram estimados, e dados clínicos obtidos junto aos prontuários dos pacientes. Dentre os isolados clínicos (n=45), C. parapsilosis sensu stricto (s.s.) representou 84%, C. orthopsilosis 16% e ausência de C. metapsilosis. Os perfis de inteins confirmaram o padrão híbrido incomum da espécie C. orthopsilosis. Dentre os ambientais (n=14), todos pertenceram à C. parapsilosis s.s. A produção de proteinase foi positiva ou fortemente positiva em 55% dos isolados clínicos de C. parapsilosis s.s., e negativa nos isolados de C. orthopsilosis. Todos os isolados ambientais de C. parapsilosis s.s. foram produtores de proteinase (64% fortemente positivos). A produção de fosfolipase foi positiva para apenas um isolado ambiental de C. parapsilosis s.s. A produção de biofilme foi significativamente maior em C. orthopsilosis. Todos os isolados clínicos de C. parapsilosis s.s. foram sensíveis à anfotericina B e voriconazol, cinco foram sensíveis dose-dependentes e dois resistentes ao fluconazol, três foram simultaneamente sensíveis dose-dependentes à caspofungina e micafungina. Os isolados de C. orthopsilosis foram sensíveis a todos os antifúngicos. A mortalidade foi alta (51%) e as mortes ocorreram principalmente em recém-nascidos e idosos. A melhor compreensão das leveduras do complexo C. parapsilosis terá impactos positivos no tratamento e prevenção de novas infecções.
Fungal infections caused by yeasts are serious problems in hospitalized patients around the globe and have become a major cause of morbidity and mortality. Although Candida albicans is the most frequently isolated species and its infection usually occurs by endogenous translocation, an increase of non-albicans species has been observed, with emphasys for the C. parapsilosis complex, whose main route of infection is exogenous, probably by the hands of health professionals. This work proposes to study the microbiological and environmental aspects of candidemia, with special attention to the C. parapsilosis complex, in a public tertiary hospital HC/FMB/UNESP, in Botucatu, using the molecular methods, active search of agents in the hospital environment (air, surfaces and hands of health professionals), as well as to determine the virulence factors and susceptibility to the antifungals of these species and correlate with the patient clinical outcomes. The clinical isolates (obtained of the blood cultures, 2007-2015 period) and environmental (2014-2015 period) of the C. parapsilosis sensu lato were identified by CHROMagar Candida medium, Vitek-2, rDNA sequencing and VMA and ThrRS inteins profiles. Virulence factors (production of proteinase, phospholypase and biofilm) and profiles of susceptibility to antifungals amphotericin B, fluconazole, voriconazole, caspofungin and micafungin were estimated, and the clinical data were obtained from patient’s records. Among the clinical isolates (n=45), C. parasilosis sensu stricto (s.s.) represented 84%, C. orthopsilosis 16% and absence of C. metapsilosis. The inteins profiles confirmed the unusual hybrid pattern of the C. orthopsilosis species. Among the environmental isolates (n=14), all belong to C. parapsilosis s.s. Proteinase production was positive or strongly positive in 55% of clinical isolates of C. parapsilosis s.s., and negative in the C. orthopsilosis isolates. All the environment isolates of C. parapsilosis s.s. were proteinase producers (64% of them were strongly positive). The phospholipase production was positive for only one environmental isolate of C. parapsilosis s.s. The biofilm production was significantly higher in C. orthopsilosis. All the clinical isolates of C. parapsilosis s.s. were sensitive to amphoterycin B and voriconazole, five were dose-dependent sensitivity and two resistant to fluconazole, three were simultaneously dose-dependent sensitivity to caspofungin and micafungin. The isolates of C. orthopsilosis were sensitive to all antifungals. The mortality was high (51%) and deaths occurred mainly in newborns and elderly patients. A better understanding of C. parapsilosis complex yeasts will have positive impacts on the treatment and prevention of new infections in our hospitals.
Fungal infections caused by yeasts are serious problems in hospitalized patients around the globe and have become a major cause of morbidity and mortality. Although Candida albicans is the most frequently isolated species and its infection usually occurs by endogenous translocation, an increase of non-albicans species has been observed, with emphasys for the C. parapsilosis complex, whose main route of infection is exogenous, probably by the hands of health professionals. This work proposes to study the microbiological and environmental aspects of candidemia, with special attention to the C. parapsilosis complex, in a public tertiary hospital HC/FMB/UNESP, in Botucatu, using the molecular methods, active search of agents in the hospital environment (air, surfaces and hands of health professionals), as well as to determine the virulence factors and susceptibility to the antifungals of these species and correlate with the patient clinical outcomes. The clinical isolates (obtained of the blood cultures, 2007-2015 period) and environmental (2014-2015 period) of the C. parapsilosis sensu lato were identified by CHROMagar Candida medium, Vitek-2, rDNA sequencing and VMA and ThrRS inteins profiles. Virulence factors (production of proteinase, phospholypase and biofilm) and profiles of susceptibility to antifungals amphotericin B, fluconazole, voriconazole, caspofungin and micafungin were estimated, and the clinical data were obtained from patient’s records. Among the clinical isolates (n=45), C. parasilosis sensu stricto (s.s.) represented 84%, C. orthopsilosis 16% and absence of C. metapsilosis. The inteins profiles confirmed the unusual hybrid pattern of the C. orthopsilosis species. Among the environmental isolates (n=14), all belong to C. parapsilosis s.s. Proteinase production was positive or strongly positive in 55% of clinical isolates of C. parapsilosis s.s., and negative in the C. orthopsilosis isolates. All the environment isolates of C. parapsilosis s.s. were proteinase producers (64% of them were strongly positive). The phospholipase production was positive for only one environmental isolate of C. parapsilosis s.s. The biofilm production was significantly higher in C. orthopsilosis. All the clinical isolates of C. parapsilosis s.s. were sensitive to amphoterycin B and voriconazole, five were dose-dependent sensitivity and two resistant to fluconazole, three were simultaneously dose-dependent sensitivity to caspofungin and micafungin. The isolates of C. orthopsilosis were sensitive to all antifungals. The mortality was high (51%) and deaths occurred mainly in newborns and elderly patients. A better understanding of C. parapsilosis complex yeasts will have positive impacts on the treatment and prevention of new infections in our hospitals.
Descrição
Palavras-chave
complexo Candida parapsilosis, fatores de virulência, identificação molecular, exposição ambiental, agentes antifúngicos, Candida parapsilosis complex, virulence factors, molecular identification, environmental exposures, antifungal agents