The bioavailability of engineered nanomaterials should be limited in marine environments, but uptake and toxicity has been noted in marine fish and invertebrates, albeit at exposure doses far exceeding predicted environmental levels. We examined the bioactivity of amine functionalized copper nanoparticles (nCu; 5-10 nm core diameter) to the euryhaline killifish, Fundulus heteroclitus, in fresh (FW) and brackish water (BW). Free copper dissolution was undetectable in either water type and nCu remained relatively well dispersed in BW, despite the high ionic strength. Exposure to an environmentally relevant concentration of nCu (10 µg L) for 48 h significantly increased the maximum rate of oxygen consumption and aerobic scope in BW killifish. This effect was associated with gill remodeling which likely increased surface area and scope for oxygen uptake. In contrast, nCu exposure had no effect on oxygen consumption in FW killifish, but gill Na/K-ATPase activity was reduced by >40%, an effect not seen in BW. Osmotic and ionic homeostasis were protected and no indications of physiological or oxidative stress were observed in either FW and BW exposure groups. The results show that functionalized nCu formulations can exhibit bioactivity in both FW and BW and that the underlying mechanisms are different between water types.