Antibacterial efficiency of surface-immobilized Flavobacterium-infecting bacteriophage

Abstract

Control of bacterial diseases by bacteriophages (phages) is gaining more interest due to increasing antibiotic resistance. This has led to technologies to attach phages on surfaces to form a biomaterial that can functionally display phages that interact with bacteria, to carry out successful infection cycles. Such a material could be applied in many environments, where the target pathogens are expected. Although this approach has been applied successfully in a few studies already, the basis of the antibacterial effect by the immobilized phages is unclear, and the interpretation of the results depends on the study. Here, we studied the phage attachment density, their detachment rate and infectivity on five different surfaces: silicon, amine-treated silicon, gold, carboxylate-treated gold and crosslinker-activated carboxylate-treated gold. The density of attached phages varied between the different surfaces and was highest on the crosslinker-activated carboxylate-treated gold. To understand whether the antibacterial effect is caused by the attached or the detached phages, the strength of the immobilization was analyzed by performing 3-12 washing steps. The detachment rates differed between the materials, with the amine treated silicon surface generating the highest release of phages and maintaining the highest infectivity, even after extensive washing. On the other hand, covalent crosslinking seemed to interfere with the infectivity. Our results suggest that the detachment of the phages from the surface is a possible mechanism for the antibacterial effect. Furthermore, we introduce a measure of the infectivity by comparing the bacterial growth reductions produced by the phage-treated materials to the effect caused by a known number of free phages, resulting in a unit “Effective PFU/surface area”, a comparable standard between different studies.