RESUMO
Although hemolytic activity is known to be a putative virulence factor contributing to candidal pathogenesis, its production by Candida tropicalis, a species closely related to Candida albicans, is poor understood. The present study was undertaken to evaluate the hemolytic activity and the expression level of a putative haem oxygenase encoding gene by blood isolates of C. tropicalis following growth in iron deprivation, and in the presence of hemoglobin and erythrocytes. The lowest values of hemolytic activity were observed in cell-free culture supernatants of isolates growing in iron-restricted medium (RPMI medium and RPMI medium supplemented with iron chelator bathophenanthrolindisulphonic acid). Hemolysis was increased in the presence of either hemoglobin or erythrocytes. Reverse transcriptase PCR analysis showed that the putative haem oxygenase encoding gene (CtHMX1), potentially related with iron uptake, was up-regulated (p < 0.001) following growth in iron deprivation and in the presence of hemoglobin; CtHMX1 was repressed in the presence of human erythrocytes (p < 0.001). Our data suggest that hemoglobin had positive effect in the production of hemolytic factor and gene expression related to iron uptake in C. tropicalis.
Assuntos
Sangue/microbiologia , Candida tropicalis/enzimologia , Candida tropicalis/genética , Eritrócitos/metabolismo , Heme Oxigenase (Desciclizante)/genética , Heme Oxigenase (Desciclizante)/metabolismo , Hemoglobinas/metabolismo , Ferro/metabolismo , Candida tropicalis/crescimento & desenvolvimento , Candida tropicalis/ultraestrutura , Candidíase/sangue , Candidíase/microbiologia , Meios de Cultura , DNA Fúngico/isolamento & purificação , Proteínas Fúngicas/biossíntese , Proteínas Fúngicas/genética , Fungos/crescimento & desenvolvimento , Proteínas Hemolisinas , Hemólise , Humanos , RNA Fúngico/isolamento & purificação , Regulação para Cima , Fatores de Virulência/metabolismoRESUMO
Candida species have been associated with the emergence of strains resistant to selected antifungal agents. Plant products have been used traditionally as alternative medicine to ease mucosal fungal infections. This study aimed to investigate the effects of Piper betle extract on the growth profile and the ultrastructure of commonly isolated oral candidal cells. The major component of P. betle was identified using liquid chromatography-mass spectrophotometry (LC-MS/MS). Seven ATCC control strains of Candida species were cultured in yeast peptone dextrose broth under four different growth environments: (i) in the absence of P. betle extract; and in the presence of P. betle extract at respective concentrations of (ii) 1 mgâ mL(-1); (iii) 3 mgâ mL(-1); and (iv) 6 mgâ mL(-1). The growth inhibitory responses of the candidal cells were determined based on changes in the specific growth rates (µ). Scanning electron microscopy (SEM) was used to observe any ultrastructural alterations in the candida colonies. LC-MS/MS was performed to validate the presence of bioactive compounds in the extract. Following treatment, it was observed that the µ-values of the treated cells were significantly different than those of the untreated cells (P<0.05), indicating the fungistatic properties of the P. betle extract. The candidal population was also reduced from an average of 13.44×10(6) to 1.78×10(6) viable cell counts (CFU)â mL(-1). SEM examination exhibited physical damage and considerable morphological alterations of the treated cells. The compound profile from LC-MS/MS indicated the presence of hydroxybenzoic acid, chavibetol and hydroxychavicol in P. betle extract. The effects of P. betle on candida cells could potentiate its antifungal activity.
Assuntos
Antifúngicos/farmacologia , Candida/efeitos dos fármacos , Boca/microbiologia , Piper betle , Extratos Vegetais/farmacologia , Candida/crescimento & desenvolvimento , Candida/ultraestrutura , Candida albicans/efeitos dos fármacos , Candida albicans/crescimento & desenvolvimento , Candida albicans/ultraestrutura , Candida glabrata/efeitos dos fármacos , Candida glabrata/crescimento & desenvolvimento , Candida glabrata/ultraestrutura , Candida tropicalis/efeitos dos fármacos , Candida tropicalis/crescimento & desenvolvimento , Candida tropicalis/ultraestrutura , Cromatografia Líquida/métodos , Contagem de Colônia Microbiana , Meios de Cultura , Eugenol/análogos & derivados , Eugenol/análise , Humanos , Hidroxibenzoatos/análise , Viabilidade Microbiana/efeitos dos fármacos , Microscopia Eletrônica de Varredura , Fitoterapia , Piper betle/química , Extratos Vegetais/análise , Espectrofotometria/métodos , Espectrometria de Massas em Tandem/métodos , Fatores de TempoRESUMO
A 6,000 Da peptide, named CaTI, was isolated from Capsicum annuum L. seeds and showed potent inhibitory activity against trypsin and chymotrypsin. The aim of this study was to determine the effect of CaTI on Saccharomyces cerevisiae, Candida albicans, Candida tropicalis and Kluyveromyces marxiannus cells. We observed that CaTI inhibited the growth of S. cerevisiae, K. marxiannus as well as C. albicans and induced cellular agglomeration and the release of cytoplasmic content. No effect on growth was observed in C. tropicalis but morphological changes were noted. In the spot assay, different degrees of sensitivity were shown among the strains and concentrations tested. Scanning electron microscopy showed that S. cerevisiae, K. marxiannus and C. albicans, in the presence of CaTI, exhibited morphological alterations, such as the formation of pseudohyphae, cellular aggregates and elongated forms. We also show that CaTI induces the generation of nitric oxide and interferes in a dose-dependent manner with glucose-stimulated acidification of the medium mediated by H(+)-ATPase of S. cerevisiae cells.
Assuntos
Antifúngicos/isolamento & purificação , Candida albicans/efeitos dos fármacos , Candida tropicalis/efeitos dos fármacos , Capsicum/enzimologia , Kluyveromyces/efeitos dos fármacos , Proteínas de Plantas/farmacologia , Saccharomyces cerevisiae/efeitos dos fármacos , Inibidores da Tripsina/farmacologia , Antifúngicos/farmacologia , Candida albicans/crescimento & desenvolvimento , Candida albicans/ultraestrutura , Candida tropicalis/crescimento & desenvolvimento , Candida tropicalis/ultraestrutura , Permeabilidade da Membrana Celular/efeitos dos fármacos , Meios de Cultivo Condicionados , Relação Dose-Resposta a Droga , Desenho de Fármacos , Avaliação Pré-Clínica de Medicamentos , Proteínas Fúngicas/antagonistas & inibidores , Glucose/farmacologia , Kluyveromyces/crescimento & desenvolvimento , Kluyveromyces/ultraestrutura , Testes de Sensibilidade Microbiana , Microscopia Eletrônica de Varredura , Óxido Nítrico/biossíntese , Proteínas de Plantas/química , Proteínas de Plantas/isolamento & purificação , ATPases Translocadoras de Prótons/antagonistas & inibidores , Saccharomyces cerevisiae/crescimento & desenvolvimento , Saccharomyces cerevisiae/ultraestrutura , Inibidores da Tripsina/química , Inibidores da Tripsina/isolamento & purificaçãoRESUMO
During olive oil production, large quantities of olive mill wastewater (OMW) are produced. This wastewater material, containing a high level of phenolic compounds, poses a serious environmental problem in almost all Mediterranean countries. Candida tropicalis YMEC14 was used as an extremophile strain to design an aerobic biotreatment process to detoxify OMW and reduce its polluting organic load. The process was enhanced by directing yeast metabolism towards biodegradation pathways using hexadecane as co-metabolite and by immobilizing yeast cells in calcium alginate beads. Under immobilization conditions, C. tropicalis YMEC14 grown at 40 degrees C in OMW supplemented with hexadecane resulted in 69.7%, 69.2% and 55.3% reduction of chemical oxygen demand, monophenols and polyphenols, respectively, after a 24-h fermentation cycle.