We conducted a study to test the hypothesis that interconnectedness among island floodplain lakes and the adjacent Solimões River during the flood stage of the hydrologic cycle is enough to maintain similarity in fish species assemblages. Gill net samples were collected during high and low water periods for three consecutive years (July 2004 to July 2006) in four lakes on Paciência Island. Two lakes, Piranha and Ressaca, are connected to the river all year, and the other two, Preto and Cacau, which are in the center of the island, are isolated during low water periods. The abundance, species richness and evenness of the fish assemblages in these lakes did not differ according to their relative positions or the season of the hydrological cycle, which confirmed our hypothesis. However, fish abundance during the dry season was greater than in the flood season. Apparently, the short period of full connection between the lakes is enough to allow the colonization of all fish species, but not to cause similar abundances. Our study indicates that persistence of the species composition of island floodplain lakes is primarily due to the annual replenishment of fish to the lakes during the flood season.