Most cases of fungal bloodstream infections (BIs) are attributed to Candida albicans; however, non-Candida albicans Candida species have recently been identified as common pathogens. Although hemolytic factor is known to be putative virulence factor contributing to pathogenicity in Candida species, its production is poorly evaluated. The present study was undertaken to analyze the production of hemolytic factor by C. albicans (10), C. tropicalis (13), and C. parapsilosis (8) isolates associated with BIs. Data of hemolysis zones on plate assay revealed that the majority of C. albicans isolates produced mild hemolytic activity whereas the majority of C. tropicalis produced strong activity. None of the tested C. parapsilosis isolates exhibited hemolysis on plate assay. We also evaluated the hemolytic activity in the cell-free broth. There were no significant differences (P > 0.05) in the secreted hemolytic activity among intra-species isolates. Different levels of secreted hemolytic factor were observed for Candida species, where C. tropicalis exhibited the highest production of hemolytic factor (P < 0.05) followed by C. albicans and C. parapsilosis. Inhibition of hemolysis (up to 89.12 %) from culture supernatant, following incubation with the lectin Concanavalin A (Con A), was observed for all three Candida species. This finding suggests that the secreted hemolytic factor of C. tropicalis and C. parapsilosis may be a mannoprotein, similar to that described for C. albicans.

Although haemolytic factor is known to be a putative virulence factor contributing to pathogenicity in Candida species, its production by Candida tropicalis is poorly understood. In this study, we analysed the culture conditions under which C. tropicalis can display haemolytic factor on plate assay and the secretion of haemolytic factor in liquid medium by clinical isolates obtained from different specimens. All the tested isolates exhibited an internal translucent ring, resembling beta-haemolysis, surrounding by a peripheral greenish-grey halo on sheep blood agar medium. Similar haemolytic pattern was observed on human blood enriched medium. Furthermore, incubation either under normal atmosphere or under increased CO(2) had no effect on haemolysis. Overall, no differences were observed on beta-haemolytic activities (P > 0.05) among tested isolates of C. tropicalis. In glucose-limited medium (RPMI 1640 with 0.2% glucose), none of the isolates induced haemolysis on red blood cells. Similarly to found on plate assays, there were no significant differences (P > 0.05) in the activity of secreted haemolytic factor in liquid medium among C. tropicalis isolates. However, after growth, the number of yeast cells varied among isolates revealing different efficiencies of haemolytic factor production. Haemolytic activity was neither inhibited by heat treatment (100 °C) nor by the addition of pepstatin A. The obtained results extend our knowledge about haemolytic factor production by Candida species.

The aim of this study was to determine in vitro haemolytic and protease activities of Candida parapsilosis and Candida tropicalis isolates, obtained from anatomically distinct sites. Analysis of haemolytic activity of C. parapsilosis and C. tropicalis isolates obtained from the same anatomic site revealed that C. tropicalis isolates from blood had statistically higher activity (P < 0.05) than C. parapsilosis. On comparison of haemolytic activities of Candida isolates obtained from different anatomic sites, C. parapsilosis isolates from tracheal secretion were found to have higher activity than blood isolates. Protease activity was detected in the majority of the isolates analysed. Analysis of proteinase activity of C. parapsilosis and C. tropicalis isolates obtained from the same anatomic site revealed that C. parapsilosis isolates from tracheal secretion had statistically higher activity than C. tropicalis isolates. On comparison of proteinase activities of Candida isolates obtained from different anatomic sites, C. parapsilosis isolates from tracheal secretion were found to have higher activity than blood and superficial lesions isolates. Furthermore, C. tropicalis isolates from superficial lesions had higher activity than tracheal secretion isolates. Our results show the potential of C. parapsilosis and C. tropicalis isolates, obtained from distinct anatomic sites, to produce haemolytic factor and proteinases. Anatomic sites of isolation seem to be correlated with these activities, particularly for C. parapsilosis isolates.

INTRODUÇÃO: Leveduras do gênero Candida são responsáveis pela maioria das infecções fúngicas em humanos. Candida tropicalis tem sido uma das mais comumente isoladas dentre as espécies não-albicans. O objetivo foi analisar a hemólise in vitro promovida por isolados clínicos de C. tropicalis provenientes de sangue e outras amostras clínicas de pacientes internados no Hospital Universitário da UEL, PR-Brasil. MÉTODOS: Foi avaliada a hemólise promovida por 28 isolados clínicos de C. tropicalis, sendo os isolados agrupados em classes de acordo com os níveis de hemólise. RESULTADOS: A maioria dos isolados de sangue apresentou hemólise fraca (+), enquanto as classes de hemólise forte (+++) e muito forte (++++) foram as predominantes nos isolados de outras amostras clínicas como urina, lesão de unha e secreção traqueal, embora não tenham sido detectadas diferenças estatísticas (p>0,05). CONCLUSÕES: Isolados de C. tropicalis, obtidos de diferentes amostras clínicas, apresentam capacidade de promover hemólise in vitro.

BackgroundCandida tropicalis is an increasingly important human pathogen associated with high mortality rates; however, little is known regarding the virulence properties of C. tropicalis, particularly the production of haemolytic factor. Although Candida spp may acquire iron from human blood red cells (RBCs) by producing a haemolytic factor that promotes cell lyses, at present there are no data regarding the effect of RBCs on the production of haemolytic molecules. The present study was undertaken to evaluate the role of human red blood cells on the production haemolytic factor by C. tropicalis; in addition, the transcription levels of a putative haemolysin-like protein gene (HLPt) were also analysed.ResultsC. tropicalis isolates produced a haemolytic factor following growth in either the absence or presence of RBCs; however, distinct levels of haemolysis were observed, with 60% of the isolates exhibiting a significant increase in the production of haemolytic factor when grown in the presence of human RBCs. All isolates in which the putative HLPt gene was up-regulated in presence of human RBCs, ranging from 1.044 to 6.965-fold, also exhibited higher haemolytic activity following growth in the presence of RBCs compared to that observed in the absence of RBCs.ConclusionsWe propose that human RBCs may induce changes in the phenotypic expression of haemolytic factor and in transcriptional levels of the putative C. tropicalis HLPt gene in an isolate-dependent fashion.