Mg(2+) can potentially influence bacterial adhesion directly through effects on electrostatic interactions and indirectly by affecting physiology-dependent attachment processes. However, the effects of Mg(2+) on biofilm structure are largely unknown. In this study, Pseudomonas fluorescens was used to investigate the influence of Mg(2+) concentration (0, 0.1 and 1.0mM MgCl(2)) on biofilm growth. Planktonic and attached cells were enumerated (based on DAPI staining) while biofilm structures were examined via confocal laser scanning microscopy and three-dimensional structures were reconstructed. Mg(2+) concentration had no influence on growth of planktonic cells but, during biofilm formation, Mg(2+) increased the abundance of attached cells. For attached cells, the influence of Mg(2+) concentration changed over time, suggesting that the role of Mg(2+) in bacterial attachment is complex and dynamic. Biofilm structures were heterogeneous and surface colonization and depth increased with increasing Mg(2+) concentrations. Overall, for P. fluorescens, Mg(2+) increased initial attachment and altered subsequent biofilm formation and structure.