Interactive effects of waterborne metals in binary mixtures on short-term gill–metal binding and ion uptake in rainbow trout (Oncorhynchus mykiss)

Metal binding to fish gills forms the basis of the biotic ligand model (BLM) approach, which has emerged as a useful tool for conducting site-specific water quality assessments for metals. The current BLMs are designed to assess the toxicity of individual metals, and cannot account for the interactive effects of metal mixtures to aquatic organisms including fish. The present study was designed mainly to examine the interactive effects of waterborne metals (Cd, Zn, Cu, Ag, and Ni) in specific binary combinations on short-term (3h) gill–metal binding and essential ion (Ca2+ and Na+) uptake (a physiological index of toxicity) in fish, using juvenile freshwater rainbow trout (Oncorhynchus mykiss) as the model species. We hypothesized that binary mixtures of metals that share a common mode of uptake and toxicity (e.g., Cd and Zn – Ca2+ antagonists, Cu and Ag – Na+ antagonists) would reduce the gill binding of each other via competitive interactions and induce less than additive effects on ion transport. In addition, the mixture of metals that have different modes of uptake and toxicity (e.g., Cd and Cu, or Cd and Ni) would not exhibit any interactive effects either on gill–metal binding or ion transport. We found that both Zn and Cu reduced gill–Cd binding and vice versa, however, Ni did not influence gill–Cd binding in fish. Surprisingly, Ag was found to stimulate gill–Cu binding especially at high exposure concentrations, whereas, Cu had no effect on gill–Ag binding. The inhibitory effect of Cd and Zn in mixture on branchial Ca2+ uptake was significantly greater than that of Cd or Zn alone. Similarly, the inhibitory effect of Cu and Ag in mixture on branchial Na+ uptake was significantly greater than that of Cu or Ag alone. The inhibitory effects of Cd and Zn mixture on Ca2+ uptake as well as Cu and Ag mixture on Na+ uptake were found to follow the principles of simple additivity. In contrast, no significant additive effect on either Ca2+ or Na+ uptake was recorded in fish exposed to the mixture of Cd and Cu. Overall, we found that although the effects of metal mixture interactions on gill–metal binding did not always match with our original assumptions, the effects of metal mixtures on toxicity in fish were generally consistent with our predictions. The findings of the present study have important implications for improving the BLM approach to assess metal mixture toxicity in fish.