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Upper Midwest Environmental Sciences Center

Aquatic Invasive Species Control

Evaluation of a Glucuronyl Transferase Assay for Predicting the Sensitivity of Species of Concern to the Lampricide TFM

Principal Investigator:

Impact of UMESC Science

The results of this research may lead to a more conscientious use of the lampricides to control lamprey populations in the Great Lakes by implementing procedures to protect lampricide sensitive non target species before a treatment.  Judicious use of lampricides to control lamprey populations in the Great Lakes is critical to sustaining the Great Lakes fishery industry.

Introduction

It is well known that the predominant mechanism for organisms to eliminate absorbed TFM is through formation of glucuronide conjugate.  Formation of a glucuronide conjugate is achieved by the enzyme uridinediphosphate gluruonyl transferase (UDP-GT).  Kane et al. (1994) determined the kinetics of the reaction between TFM and this enzyme for bluegills, rainbow trout, catfish, and sea lamprey.  Their research demonstrated that the tolerance of these fish toward TFM is related to the UDP-GT enzyme activity of the various species. It may be possible to refine this technique into a method for estimating the sensitivity of fish to TFM treatments. One area in which this would be particularly useful is in estimating the impacts of TFM treatments to species of concern, particularly threatened or endangered species. Because of the large number of individuals required for toxicological assays, a surrogate species is often used to estimate the toxicity to the species of concern.  The toxicological response of the surrogate, however, may not be the same as that of the species of concern. An assay for UDP-GT activity would require only a few individuals of a species of concern and would provide data directly on the organism rather than inferring impacts based on surrogate data. Data on the enzyme activity of the species of concern would be compared to data developed on the enzyme activity for species who’s sensitivity to TFM has already been established. Finally, if this approach can be developed into a useful technique for fish, it may also be expanded to include amphibians and terrestrial wildlife that utilize glucuronidation as the major pathway for elimination of TFM from their bodies.  This study will modify a UDP-GT assay for TFM in fish liver then validate the performance of the assay. The assay will be capable of determining the UDP-GT enzyme activity in a variety of fish liver found in the Great Lakes drainage basin.

Objectives

  1. Modify and validate an enzymatic assay for the conversion of 3-trifluoromethyl-4-nitrophenol (TFM) to TFM glucuronide in fish liver.  The assay will determine the uridinediphosphate gluruonyl transferase (UDP-GT) activity per gram of fish liver (formation of TFM glucuronide / gram liver /minute at 20 ℃).
  2. Determine if the assay results can predict the relative sensitivity of fish species to 3-trifluoromethyl-4-nitrophenol (TFM).

Reference

Kane, A.S, M.W. Kahng, and R. Reimschuessel.  1994.  UDP-Glucuronyltransferase Kinentics for 3-Trifluoromethyl-4-nitrophenol (TFM) in Fish.  Transactions of the American Fisheries Society 123: 217-222.

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