The current concept of ventricular filling in elasmobranch and teleost fishes is that atrial contraction is the primary, if not the exclusive, determinant of ventricular filling. Recent echocardiographic and on-line hemodynamic data for elasmobranchs, however, have demonstrated a biphasic ventricular filling pattern, characterized by an early phase that occurs during ventricular relaxation and a late phase that follows atrial systole. This study reports echocardiographic and hemodynamic analyses of ventricular filling in three teleost genera (Paralabrax, Channa, Monopterus) having markedly different heart morphologies. Both the profiles of the atrioventricular pressure gradient in Paralabrax and the ventricular inflow velocity in all three genera indicate a biphasic ventricular filling pattern. Although the relative contribution of the early and late filling phases differed among the species studied, interspecific differences in heart structure did not obscure the biphasic pattern. Also, pericardiectomy did not affect the biphasic ventricular filling pattern in Paralabrax. The presence of biphasic filling in teleosts establishes a functional similarity with the elasmobranchs and, because the biphasic ventricular filling pattern predominates in higher vertebrates, suggests that this ventricular filling mechanism may be present in the entire subphylum Vertebrata.