Enzymatic analysis of liver samples from rainbow trout for diagnosis of blue-green algae-induced toxicosis

Ali Sahin From the Institute of Veterinary Pharmacology and Toxicology, University of Zurich, Winterthurerstr 260, 8057 Zurich (Sahin, Naegeli); Institute of Plant Biology, University of Zurich, Zollikerstr 107, 8008 Zurich (Mez); and Institute of Toxicology, Federal Institute of Technology and University of Zurich, Schorenstr 16, 8603 Schwerzenbach (Tencalla, Dietrich), Switzerland. A. Sahin is a research assistant at the Department of Internal Diseases and Pharmacology, University of Yüzüncü Yil, Van, Turkey.

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Francesca G. Tencalla From the Institute of Veterinary Pharmacology and Toxicology, University of Zurich, Winterthurerstr 260, 8057 Zurich (Sahin, Naegeli); Institute of Plant Biology, University of Zurich, Zollikerstr 107, 8008 Zurich (Mez); and Institute of Toxicology, Federal Institute of Technology and University of Zurich, Schorenstr 16, 8603 Schwerzenbach (Tencalla, Dietrich), Switzerland. A. Sahin is a research assistant at the Department of Internal Diseases and Pharmacology, University of Yüzüncü Yil, Van, Turkey.

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Daniel R. Dietrich From the Institute of Veterinary Pharmacology and Toxicology, University of Zurich, Winterthurerstr 260, 8057 Zurich (Sahin, Naegeli); Institute of Plant Biology, University of Zurich, Zollikerstr 107, 8008 Zurich (Mez); and Institute of Toxicology, Federal Institute of Technology and University of Zurich, Schorenstr 16, 8603 Schwerzenbach (Tencalla, Dietrich), Switzerland. A. Sahin is a research assistant at the Department of Internal Diseases and Pharmacology, University of Yüzüncü Yil, Van, Turkey.

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Konstanze Mez From the Institute of Veterinary Pharmacology and Toxicology, University of Zurich, Winterthurerstr 260, 8057 Zurich (Sahin, Naegeli); Institute of Plant Biology, University of Zurich, Zollikerstr 107, 8008 Zurich (Mez); and Institute of Toxicology, Federal Institute of Technology and University of Zurich, Schorenstr 16, 8603 Schwerzenbach (Tencalla, Dietrich), Switzerland. A. Sahin is a research assistant at the Department of Internal Diseases and Pharmacology, University of Yüzüncü Yil, Van, Turkey.

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Hanspeter Naegeli From the Institute of Veterinary Pharmacology and Toxicology, University of Zurich, Winterthurerstr 260, 8057 Zurich (Sahin, Naegeli); Institute of Plant Biology, University of Zurich, Zollikerstr 107, 8008 Zurich (Mez); and Institute of Toxicology, Federal Institute of Technology and University of Zurich, Schorenstr 16, 8603 Schwerzenbach (Tencalla, Dietrich), Switzerland. A. Sahin is a research assistant at the Department of Internal Diseases and Pharmacology, University of Yüzüncü Yil, Van, Turkey.

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SUMMARY

Microcystin and related toxic peptides produced by cyanobacteria (blue-green algae) are potent and selective inhibitors of protein phosphatases 1 and 2A. We adapted existing enzymatic techniques to analyze the liver of rainbow trout after oral administration of hepatotoxic cyanobacteria. Liver tissue was removed 3 and 12 hours after treatment, and phosphatase activity was determined in liver extracts, using a specific phosphoprotein substrate. In all samples from fish exposed to toxic cyanobacteria, phosphatase activity was suppressed, whereas the control enzyme, lactate dehydrogenase, present in the same liver extract, was not affected by cyanobacteria. Thus, experimental poisoning by hepatotoxic cyanobacteria resulted in an abnormally low ratio of phosphatase to lactate dehydrogenase activity in the liver extracts. These results indicate that specific inhibition of phosphatases 1 and 2A may provide a useful diagnostic tool to determine the early effects of cyanobacteria toxic peptides directly in liver samples from poisoned animals. Although this test was developed with rainbow trout, it should be possible to extend the analysis of liver phosphatase activity to other species, including sheep and cattle, which are frequently affected by hepatotoxic cyanobacteria.

SUMMARY

Microcystin and related toxic peptides produced by cyanobacteria (blue-green algae) are potent and selective inhibitors of protein phosphatases 1 and 2A. We adapted existing enzymatic techniques to analyze the liver of rainbow trout after oral administration of hepatotoxic cyanobacteria. Liver tissue was removed 3 and 12 hours after treatment, and phosphatase activity was determined in liver extracts, using a specific phosphoprotein substrate. In all samples from fish exposed to toxic cyanobacteria, phosphatase activity was suppressed, whereas the control enzyme, lactate dehydrogenase, present in the same liver extract, was not affected by cyanobacteria. Thus, experimental poisoning by hepatotoxic cyanobacteria resulted in an abnormally low ratio of phosphatase to lactate dehydrogenase activity in the liver extracts. These results indicate that specific inhibition of phosphatases 1 and 2A may provide a useful diagnostic tool to determine the early effects of cyanobacteria toxic peptides directly in liver samples from poisoned animals. Although this test was developed with rainbow trout, it should be possible to extend the analysis of liver phosphatase activity to other species, including sheep and cattle, which are frequently affected by hepatotoxic cyanobacteria.

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