Juvenile rainbow trout acclimated to softwater were exposed to 0 or 8.3 nM Ag (added as silver nitrate) for 21 days. On days 11 7 and 21 gill, kidney and liver levels of silver; branchial Na+ influx, efflux and net flux rate; gill and kidney K+-dependent P-nitrophenol phosphatase activity; and gill and liver accumulation of "new" Ag were measured. In addition, the concentration-dependent uptake of Ag by gill basolateral membrane vesicles (BLMV) was assessed in control fish and those exposed to 8.3 nM Ag for 7 days. Ag induced a significant increase in Na+ efflux following I day of exposure that resulted in an increase in net loss of Na+ and a reduction in Na+ influx. By day 21 this perturbation to Na+ balance had been corrected, but kidney K+-dependent p-nitrophenol phosphatase activity was significantly reduced. Unexpectedly, the Ag concentrations in the liver of Ag exposed fish only significantly increased (two-fold) following 7 days of exposure and were not elevated when compared to controls on day 21. In contrast, the gill and kidney accumulated significant concentrations of Ag (20-fold increase) following 7 days of exposure, and the Ag concentration in these tissues remained similar on day 21. The gills of Ag exposed fish accumulated significantly less "new" Ag than the controls on days 7 and 21 following exposure, suggesting a down-regulation of branchial Ag uptake. The BLMV of Ag exposed fish showed a significant increase in V,,,, [control fish BLMV V-max = 2811.9 +/- 190.8pmol Ag-110m/(mg protein x min) and Ag exposed fish BLMV V-max = 3688.3 +/- 659.8 pmol Ag-110m/(mg protein x min) (P = 0.033)], suggesting that they are able to increase export of Ag from the gills on exposure to Ag. The results from this study demonstrate a complex array of physiological processes that control the bioreactive concentrations of Ag in the gills, including: cytoplasmic sequestration, a down-regulation of apical entry and potentially an increase in basolateral membrane extrusion. (c) 2004 Elsevier B.V. All rights reserved.