Enterococcus faecalis thrives in dual-species biofilm models under iron-rich conditions.

Escherichia coli (E. coli) and Enterococcus faecalis (E. faecalis) are pathogenic strains that often coexist in intestinal flora of humans and are prone to cause biofilm-associated infections, such as gastrointestinal tract and urinary tract infections. Earlier studies have demonstrated that E. faecalis biofilm can metabolize ferrous ions in iron-rich environments and promote biofilm growth under in-vivo conditions. However, the influence of iron transporters on dual-species biofilm growth and the nature of molecular-level interactions between iron transporter proteins and Fe2+ remains unknown. Therefore, in this work, co-culture studies were performed and the study indicates that Fe2+ at concentrations of 50-150 µM promotes the colonization of E. coli, and Fe2+ concentrations of 50-200 µM promote the growth of E. faecalis and dual-species colonies. Atomic absorption spectroscopy results reveal that Fe2+ ion augmentation in bacterial cells was increased to 4 folds in the single-species model and 11 folds in the dual-species model under iron-supplemented conditions. Furthermore, Fe2+ augmentation increased the antibiotic resistance of E. faecalis in both single- and dual-species bacterial cultures. In addition, in-silico docking were performed to determine a three-dimensional (3D) structure of ferrous iron-transporter proteins FeoB of E. faecalis and its affinity to extracellular Fe2+. Our model suggests that the FeoB facilitates the Fe2+ uptake in E. faecalis cells in the absence of iron chelator, 2,2-bipyridyl.

Survival strategy of Pseudomonas aeruginosa on the nanopillar topography of dragonfly (Pantala flavescens) wing.

Discovery of nanopillars on the surface of the insect wings had led to the understanding of its bactericidal property. Nanopillar topography is deterrent to only those bacteria that are attached, or in close contact with the nanopillars. The present study investigated the variation in the viability of Pseudomonas aeruginosa strains PAO1 (virulent) and ATCC 9027 (avirulent) on the wing surface of dragonfly (Pantala flavescens). Viability study indicated that only 0.2% ATCC 9027 survived when incubated with wing for 48 h in Phosphate buffered saline, while under the same conditions 43.47% PAO1 survived. Enumeration of Pseudomonas attached to wing surface suggested that, the number of PAO1 attached on the wing surface was three times lesser than ATCC 9027. Propensity of attachment of P. aeruginosa strains PAO1 and ATCC 9027 on the wing surface investigated using scanning probe microscope indicated that P. aeruginosa ATCC 9027 showed adhesion to 88% of regions and, PAO1 showed adhesion to only 48% regions tested on wing surface. PAO1 survived the bactericidal effect of wing surface by evading attachment. Three clinical isolates tested which showed viability similar to PAO1 strain, also showed lower propensity to attach to wing surface. Transcriptional level analyses using RT-PCR suggested that flagellar genes (fliE and fleS) were downregulated and genes responsible for reversible to irreversible attachment (gcbA and rsmZ) were upregulated in ATCC 9027 than PAO1 on wing surface, indicating relatively higher attachment of ATCC 9027 on wing surface. The study suggests that virulent strains of P. aeruginosa may evade attachment on wing surface. The results gain significance as bioinspired surfaces are being created towards developing antibacterial medical implants and other antibacterial surface applications.