With the increasing use of silver nanoparticles (Ag-NPs), their entrance into aquatic ecosystems is inevitable. Thus, the present study simulated the potential fate, toxicity, and bioaccumulation of Ag-NPs released into aquatic systems with different salinities. The Ag-NPs were characterized using inductively coupled plasma-atomic emission spectroscopy (ICP-AES), dynamic light scattering (DLS), transmission electron microscopy (TEM), energy-dispersive X-ray analysis (EDX), and UV-vis spectroscopy. Juvenile rainbow trout were exposed to Ag-NPs in three different salinity concentrations, including low (0.4 ppt), moderate (6 ± 0.3 ppt), and high (12 ± 0.2 ppt) salinity, for 14 days in static renewal systems. The nominal Ag-NP concentrations in the low salinity were 0.032, 0.1, 0.32, and 1 ppm, while the Ag-NP concentrations in the moderate and high salinity were 3.2, 10, 32, and 100 ppm. UV-vis spectroscopy was used during 48 h (re-dosing time) to evaluate the stability and possible changes in size of the Ag-NPs in the water. The results revealed that the λmax of the Ag-NPs remained stable (415-420 nm) at all concentrations in the low salinity with a reduction of absorbance between 380 and 550 nm. In contrast, the λmax quickly shifted to a longer wavelength and reduced absorbance in the moderate and higher salinity. The bioaccumulation of Ag in the studied tissues was concentration-dependent in all the salinities based on the following order: liver>kidneys≈gills>white muscles. All the tissue silver levels were significantly higher in the high salinity than in the moderate salinity. In addition, all the fish exposed to Ag-NPs in the low, moderate, and high salinity showed a concentration-dependent increase in their hepatosomatic index (HSI). In conclusion, most Ag-NPs that enter into freshwater ecosystems (low ionic strength) remain suspended, representing a potentially negative threat to the biota in an ionic or nanoscale form. However, in a higher salinity, nanoparticles agglomerate and precipitate on the surface of the sediment.