The presence of aquatic macrophytes is a key factor in the selection of habitats by fish in floodplain lakes because these plants enhance the physical and biological complexities of aquatic habitats. The seasonal flood pulse may influence this interaction, but there is no information in the literature about the effects that flood events may have on macrophytes assemblages and its associated effects on fish assemblages. Thus, this article aimed to investigate whether species richness, evenness and similarities in fish assemblage composition differed between littoral areas vegetated with macrophytes and unvegetated areas, before and after a flood. We sampled three lakes in the floodplain of the upper Paraná River basin. Sampling was conducted before (December 2004 and January 2005) and after (early March, late March and May 2005) a flood event. Overall, species richness and evenness were higher in macrophytes-covered areas. Before the flood, the composition of fish assemblages was distinct when comparing vegetated and unvegetated areas. After the flood, the similarity in fish assemblage composition was higher, indicating a homogenization effect of floods for fish inhabiting littoral areas of floodplain lakes. After the flood, opportunistic species dominated the fish assemblages in aquatic macrophytes, apparently restructuring assemblages in the littoral, restarting a succession process. Thus, the observed homogenization effect of the flood could minimize biological interactions and could induce fish assemblages to begin a new process of structurization.