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Validation of a cell culture bioassay for detection of petroleum exposure in mink (Mustela vison) as a model for detection in sea otters (Enhydra lutris)

Michael H. ZiccardiWildlife Health Center, University of California, Davis, CA 95616.
Pathology, Microbiology, and Immunology, University of California, Davis, CA 95616.

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 DVM, MPVM, PhD
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Jonna A. K. MazetWildlife Health Center, University of California, Davis, CA 95616.
Department of Medicine and Epidemiology, University of California, Davis, CA 95616.

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Ian A. GardnerWildlife Health Center, University of California, Davis, CA 95616.
Department of Medicine and Epidemiology, University of California, Davis, CA 95616.

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Walter M. BoyceWildlife Health Center, University of California, Davis, CA 95616.
Pathology, Microbiology, and Immunology, University of California, Davis, CA 95616.

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Michael S. DenisonDepartment of Environmental Toxicology, University of California, Davis, CA 95616.

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 PhD

Abstract

Objective—To validate a luciferase bioassay, which is based on a recombinant mouse hepatoma cell line, for the detection of exposure to petroleum in mustelid species.

Animals—122 American mink (Mustela vison) and 15 sea otters (Enhydra lutris).

Procedures—Mink were exposed to Bunker C fuel oil or Alaska North Slope crude oil externally as a single exposure or internally via low dose concentrations in their ration for 6 months. Serum samples were analyzed for cytochrome P450 1A1 induction by quantification of luciferase activity in the bioassay. Mink liver specimens were also evaluated for cytochrome P450 1A1 induction by quantification of ethoxyresorufin-o-deethylase activity. Serum collected from exposed and unexposed sea otters was also analyzed using the luciferase bioassay.

Results—Serum samples from mink externally exposed to petroleum had significantly increased luciferase activities at 1 week after exposure. Serum samples taken at later time points or from mink exposed to either product in the ration did not cause significant luciferase induction. Samples from otters exposed to petroleum had significantly higher luciferase induction as compared with samples from otters not exposed to petroleum at 2 and 8 years after the spill. Cytochrome P450 1A1 activity in liver specimens collected from mink that were internally exposed through diet was significantly increased at the conclusion of our study.

Conclusion and Clinical Relevance—The luciferase bioassay is a sensitive and specific method for determining recent exposure to petroleum in mink. The lack of luciferase activity in serum samples collected from mink greater than 1 week after experimental exposure was likely attributable to lower overall petroleum exposure in our trial, compared with natural exposures. (Am J Vet Res 2002;63:963–968)

Abstract

Objective—To validate a luciferase bioassay, which is based on a recombinant mouse hepatoma cell line, for the detection of exposure to petroleum in mustelid species.

Animals—122 American mink (Mustela vison) and 15 sea otters (Enhydra lutris).

Procedures—Mink were exposed to Bunker C fuel oil or Alaska North Slope crude oil externally as a single exposure or internally via low dose concentrations in their ration for 6 months. Serum samples were analyzed for cytochrome P450 1A1 induction by quantification of luciferase activity in the bioassay. Mink liver specimens were also evaluated for cytochrome P450 1A1 induction by quantification of ethoxyresorufin-o-deethylase activity. Serum collected from exposed and unexposed sea otters was also analyzed using the luciferase bioassay.

Results—Serum samples from mink externally exposed to petroleum had significantly increased luciferase activities at 1 week after exposure. Serum samples taken at later time points or from mink exposed to either product in the ration did not cause significant luciferase induction. Samples from otters exposed to petroleum had significantly higher luciferase induction as compared with samples from otters not exposed to petroleum at 2 and 8 years after the spill. Cytochrome P450 1A1 activity in liver specimens collected from mink that were internally exposed through diet was significantly increased at the conclusion of our study.

Conclusion and Clinical Relevance—The luciferase bioassay is a sensitive and specific method for determining recent exposure to petroleum in mink. The lack of luciferase activity in serum samples collected from mink greater than 1 week after experimental exposure was likely attributable to lower overall petroleum exposure in our trial, compared with natural exposures. (Am J Vet Res 2002;63:963–968)