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1. Liver cytosol samples from juvenile coho salmon exposed over a 14-week period to 70 micrograms/l (1/4 LC50) or 140 micrograms/l (1/2 LC50), were separated into high-molecular-weight and low-molecular-weight fractions by column chromatography on Sephadex G-75, and compared to similar samples obtained from control fish. 2. The levels of copper in the low-molecular-weight function of fish exposed to 70 micrograms/l were not significantly increased over control values for the first 6 weeks but then increased rapidly, while those of fish exposed to 140 micrograms/l increased substantially in the first 2-4 weeks of exposure, then levelled off. 3. In the exposed fish the levels of copper in the high-molecular-weight fraction were not significantly elevated above control levels over the first 8-10 weeks but then increased significantly. 4. The low-molecular-weight fractions of exposed fish were shown to contain increasing levels of metallothionein. The metallothionein was isolated on DEAE-cellulose and characterized by amino acid analyses.

Juvenile coho salmon (Oncorhynchus kisutch) 3-8 g body weight were exposed to sublethal concentrations of copper acetate in water of hardness 280 mg/l (as CaCO3) using flow-through conditions. Concentrations of copper were approximately 1, 50, 100 or 150 micrograms Cu/l. Uptake of copper into blood, gills, kidney and liver was measured using 67Cu and into hepatic metallothionein. Chromatography on Sephadex G-75 was used to separate the protein from the cytosol. Copper was determined using atomic absorption spectrophotometry. The ratio of rates of uptake of copper and synthesis of metallothionein was consistent with published values for the copper content of copper-thionein, i.e. 7 to 1. After 4 weeks of continuous exposure the concentrations of copper and hepatic metallothionein levelled off at concentrations that were a function of the copper concentration in the water. After the plateau or steady-state had been reached, the rate of loss of metallothionein was measured by determining the change in specific-activity of isotope with time after incorporation of 35S-L-cysteine or 3H-L-lysine into the protein. The half-life for loss of metallothionein was a function of the exposure to copper, being about 13 days for control fish and 30 days for fish exposed to the greatest concentration of copper. Salmon that had been exposed to copper for 8 weeks neither gained hepatic metallothionein in a further 4 week exposure to copper, nor lost it when transferred for 4 weeks to clean water. Tolerance to copper exhibited by fish that continued to be exposed to it was partially lost after 4 weeks depuration.(ABSTRACT TRUNCATED AT 250 WORDS)

Rainbow trout developed elevated hepatic metallothionein concentrations after 4 weeks in a solution containing zinc, copper and cadmium in a fixed ratio of 400:20:1. Resistance to a combination of these metals increased in proportion to the concentration to which they were exposed for 4 weeks. The concentration of copper but not zinc or cadmium in the low molecular weight proteins separated by gel filtration was related to the concentrations of metallothionein present. The combined toxicity of the metals in the mixture was additive.

Rainbow trout held in net pens for 4 weeks in three locations contaminated by heavy metals developed elevated levels of hepatic metallothionein but no acclimation to a combination of zinc, copper and cadmium. Toxicity of the mixture of zinc, copper and cadmium in a ratio of 400:20:1 increased with temperature in the range of 6-21 degrees C. Hepatic metallothionein concentrations after 4 weeks of exposure in situ was correlated with the degree of contamination as measured by zinc concentrations (P less than 0.01).

1. Four groups of 600 juvenile coho salmon (Oncorhynchus kisutch) of 3-8g body weight were exposed to sublethal concentrations of copper acetate in water of hardness 280 mg/l (as CaCO3). 2. Flow-through conditions were maintained for a period of 10 weeks, during which the copper concentrations to which the groups of fish were exposed were approximately 1, 50, 100 and 150 micrograms Cu/l. 3. Hepatic metallothionein concentrations were measured using differential pulse polarography in filtrates of heat-denatured saline extracts of the tissues. 4. These values increased as a function of the copper concentration to maximum levels after 4 weeks of continuous exposure and thereafter remained more or less constant. 5. Flow-through 168 hr LC50 values for fish tested at intervals throughout the experiment increased to maximum values (2.5 X control values) at 1 week and during the next 3 weeks fell somewhat to stable but elevated values. 6. At 4 weeks, these were related to the metallothionein (MT) values by the regression equation 168 hr LC50 = 2.423 (+/- 0.455) X MT + 48.6 (+/- 31.1). 7. The probability that the relationship was not accidental exceeded 97%.