Zinc oxide nanoparticles (ZnO NPs) are increasingly used in several personal care products, with high potential to be released directly into marine environment with consequent adverse impact on marine biota. This paper aimed to compare the ecotoxicological effect of ZnO NPs (< 100 nm) towards three marine organisms widely used in toxicity assessment: an algal species (Dunaliella tertiolecta), a bioluminescent bacterium (Vibrio fischeri), and a crustacean (Artemia salina). Bulk ZnO (ZnO bulk, 200 nm) and ionic zinc were also investigated for understanding the role of size and of ionic release in the ZnO toxic action. To this aim, different ecotoxicological tests were used: the inhibition of bioluminescence with V. fischeri at three exposure times (5, 15, and 30 min); the D. tertiolecta growth inhibition at 24, 48, and 72 h; the A. salina mortality at 24-96 h, and A. salina mortality and body growth each 3 days along chronic exposure (14 days). For all selected species, ZnO NPs toxicity was strictly dependent on the exposure time and different sensitivities were recorded: ZnO NPs were more toxic towards algae (EC 2.2 mg Zn/L) but relatively less toxic towards bacteria (EC 17 mg Zn/L) and crustaceans (EC 96 h 58 mg Zn/L). During the 14-day chronic exposure of A. salina, ZnO NPs had a significant inhibition of vitality and body length (EC14d 0.02 mg Zn/L), while the effect of ZnSO was not statistically different from the control. ZnO NP toxicity was related to zinc ions and to interactions of particle/aggregates with target organisms and therefore to NP behavior in the testing matrix and to the different testing time exposures.