Distribution of Ebp pili among clinical and fecal isolates of Enterococcus faecalis and evaluation for human platelet activation.

Although Enterococcus faecalis is known as normal flora in colon, it is also amongst the most common causative agents of infective endocarditis (IE). Platelet activation resulting from adherence to platelets is an essential step in the pathogenesis of IE. One of the factors proposed in adhesion is endocarditis- and biofilm- associated pili encoded by ebp operon. The aim of this study was to investigate ebp in isolates from different origins and analyze the potential of isolates to activate human platelets of different donors. The ebp distribution was investigated in E. faecalis from different origin infections (n = 103) and fecal flora (n = 20). Then, selected isolates from blood (n = 5), urine (n = 2), and fecal flora (n = 3) were analyzed by flow cytometry assay for the ability to activate platelets of four different donors. No statistically significant difference was found for the ebp presence between infective and fecal isolates. Also, it was found that the ability for platelet activation is independent of the bacterial origin. However, significant difference was found in platelet activation between different donors. The results suggest that the presence or absence of ebp is not a critical factor for platelet activation by E. faecalis isolates. However, host factors seem to contribute in this activity.

Survey for Correlation between Biofilm Formation and Virulence Determinants in a Collection of Pathogenic and Fecal Enterococcus faecalis Isolates.

BACKGROUND: Enterococcus faecalis is an opportunistic pathogen that causes most of the enterococcal infections. Among the different factors implicated in the pathogenesis of these organisms, biofilm formation and antibiotic resistance are the most important. The ability for biofilm formation has been attributed to the presence of some virulence genes. However, no definite correlation has been found. This study aimed to detect biofilm formation and antibiotic resistance patterns in E. faecalis isolates collected from clinical and fecal samples, and to investigate possible correlation between some virulence genes (esp, cyl, gelE) and biofilm formation. MATERIALS AND METHODS: A collection of 123 E. faecalis isolates were investigated for antibiotic resistance and production of hemolysin, gelatinase, and biofilm using phenotypic methods. The esp, gelE and cyl genes were detected using polymerase chain reaction. RESULTS: Thirty-eight pathogenic isolates (37%) were positive for biofilm formation. Additionally, the gelE, esp, and cyl genes were detected in 74 (71.8%), 79 (76.7%) and 42 (40.8%) isolates, respectively. In the fecal samples, 18 (90%) isolates were biofilm producers and 11 (55%), 17 (85%) and 8 (40%) isolates were positive for gelE, esp, and cyl, respectively. There were significant differences in biofilm production between pathogenic and fecal isolates (P <0.001). Multidrug resistance (MDR) was found among 32% (n = 33) and 15% (n = 3) of the clinical and fecal isolates, respectively. However, no significant difference was seen between MDR and biofilm formation. Five pathogenic and two fecal isolates were negative for all investigated genes while they were they were biofilm producers. In contrast, 22 pathogenic isolates and 1 fecal isolate were positive for the tested genes, but did not form any biofilm. No significant differences were observed between biofilm formation and the presence of the esp, gelE and cyl genes in the pathogenic and fecal isolates (P >0.05). CONCLUSION: The presence of the esp, gelE and cyl genes might not be determining factors for biofilm formation in enterococci and other mechanisms might be involved in this process.