Oxygen and carbon isotopic composition of the aragonite of fish otoliths was measured on 175 specimens comprising 24 different species in 1989 and 1990. All specimens but two came from the northern Adriatic Sea or the northern Tyrrhenian Sea (two freshwater specimens were studied for comparison with the marine fish). The data obtained confirm the results of previous research suggesting the existence of equilibrium conditions between the otolith aragonite and ambient water with respect to ·18O(CO3 2-) values. Examination of one of the species indicated that the CaCO3 of otoliths probably accumulates continuously over time, seasonal isotopic changes being clearly visible (from a set of radial spot samples) for both oxygen and carbon isotopes. The apparent isotopic equilibrium with “ambient” water suggests that the δ18O(H2O) of the endolymph is equal to that of seawater and considerably different from that of fish body water. In the case of δ13C(CO3 2-), isotopic equilibrium with dissolved carbon species in seawater is never reached, even though the contribution of metabolic CO2 is variable among different species and even among different individuals of the same species. This rules out the possibility of using δ13C(CO3 2-) values obtained from fossil otoliths for paleoenvironmental and paleobiological conclusions.