Biofilm formation at oil-water interfaces is not a simple function of bacterial hydrophobicity.

Bacterial adsorption to interfaces is the initial step in biofilm formation. The mechanism of biofilm formation at liquid-liquid interfaces differs from the process of biofilm formation on solid-liquid interfaces. Until now, the former is not well understood. We study the bacterial adsorption and biofilm formation of three different bacteria, P. aeruginosa, S. aureus, and S. epidermidis at the n-decane-water interface, with focus on the relationship between bacteria wettability, excretion of biosurfactants, and biofilm formation. The adhesion capacity of these bacteria to hydrocarbons was characterized using the bacterial adherence to hydrocarbons test. We monitored the interfacial rheology of bacterial adsorption and biofilm formation at the interface over time using a drop shape analyzer and imaged the formed biofilms by using fluorescence and scanning electron microscopy. P. aeruginosa showed high adhesion capacity to hydrocarbons, while the adhesion capacity of both staphylococci was negligible. P. aeruginosa also showed rapid adsorption to the n-decane-water interface as measured by the rapid decrease in interfacial tension for the pure bacteria suspension. However, S. epidermidis, with a negligible hydrophobicity value, showed the most substantial reduction in interfacial tension and the formation of the most elastic biofilms at the oil-water interface. S. epidermidis accomplishes this by the secretion of biosurfactants. S. aureus did not form biofilms at the n-decane-water interface, in contrast to P. aeruginosa and S. epidermidis. We conclude that bacterial adsorption and biofilm formation at oil-water interfaces, in general, are not simple functions of cell hydrophobicity. Biosurfactant modification and metabolism of the interface also play essential roles.