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description: A die-off of freshwater mussels in 1990, attributed to anticholinesterase pesticide contamination of a North Carolina stream, has led the National Biological Service and the U. S. Fish and Wildlife Service to explore the development of biomonitoring programs using cholinesterase activity to assess the threat of anticholinesterase pesticides to freshwater mussels. However, background information such as "normal" cholinesterase activities and basic biochemical properties of the cholinesterases present in mussels is extremely limited. Early attempts to identify baseline cholinesterase activities for field-collected eastern elliptio (Elliptio complanata) have been plagued by high levels of variation in activities measured in mussels exposed to the same environmental conditions. The objectives of this study were two-fold: 1) to elucidate and reduce this variability through the characterization of the cholinesterases involved and the refinement of assay protocols, and 2) to continue for a second year the biomonitoring of cholinesterase activities in E. complanata in the area of the Tar River basin, North Carolina, where the die-off occurred. Enzyme characterization studies discovered that cholinesterase activities in crude homogenates of adductor muscle demonstrated inhibition with increasing substrate concentrations, preferred acetylthiocholine as a substrate over butyryl- and propionylthiocholine, and was not significantly inhibited by a specific butyrylcholinesterase inhibitor, suggesting that the predominately active enzyme in adductor muscle tissue is acetylcholinesterase. Minor improvements to the assay protocols did not lower the overall variation in activities of field samples. Coefficients of variation for each collection event still ranged from 24.12% to 65.46%. However, despite the large intra-site variation in ChE activities, the Hilliardston collection site, located near the 1990 die-off site, did reveal a significantly (p < 0.05) lower average cholinesterase activity (94.41 + 46.05 umoles substrate hydrolyzed/min/g protein) than the Berea reference site (141.20 + 58.36 umoles substrate/min/g protein). Evidence was insufficient to conclude that the decrease in cholinesterase activity at the die-off site was due to anticholinesterase agents. It is suggested that the variation in cholinesterase activities is mainly influenced by characteristics of the individual rather than measured water quality parameters. These characteristics are discussed along with recommendations for improving the biomonitoring program.; abstract: A die-off of freshwater mussels in 1990, attributed to anticholinesterase pesticide contamination of a North Carolina stream, has led the National Biological Service and the U. S. Fish and Wildlife Service to explore the development of biomonitoring programs using cholinesterase activity to assess the threat of anticholinesterase pesticides to freshwater mussels. However, background information such as "normal" cholinesterase activities and basic biochemical properties of the cholinesterases present in mussels is extremely limited. Early attempts to identify baseline cholinesterase activities for field-collected eastern elliptio (Elliptio complanata) have been plagued by high levels of variation in activities measured in mussels exposed to the same environmental conditions. The objectives of this study were two-fold: 1) to elucidate and reduce this variability through the characterization of the cholinesterases involved and the refinement of assay protocols, and 2) to continue for a second year the biomonitoring of cholinesterase activities in E. complanata in the area of the Tar River basin, North Carolina, where the die-off occurred. Enzyme characterization studies discovered that cholinesterase activities in crude homogenates of adductor muscle demonstrated inhibition with increasing substrate concentrations, preferred acetylthiocholine as a substrate over butyryl- and propionylthiocholine, and was not significantly inhibited by a specific butyrylcholinesterase inhibitor, suggesting that the predominately active enzyme in adductor muscle tissue is acetylcholinesterase. Minor improvements to the assay protocols did not lower the overall variation in activities of field samples. Coefficients of variation for each collection event still ranged from 24.12% to 65.46%. However, despite the large intra-site variation in ChE activities, the Hilliardston collection site, located near the 1990 die-off site, did reveal a significantly (p < 0.05) lower average cholinesterase activity (94.41 + 46.05 umoles substrate hydrolyzed/min/g protein) than the Berea reference site (141.20 + 58.36 umoles substrate/min/g protein). Evidence was insufficient to conclude that the decrease in cholinesterase activity at the die-off site was due to anticholinesterase agents. It is suggested that the variation in cholinesterase activities is mainly influenced by characteristics of the individual rather than measured water quality parameters. These characteristics are discussed along with recommendations for improving the biomonitoring program.
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Title Cholinesterase activity as a device for biomonitoring pesticide exposure in the freshwater mussel Elliptio complanata.
creation  Date   2018-05-19T15:59:30.985857
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URL:https://ecos.fws.gov/ServCat/DownloadFile/21550?Reference=23031
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Metadata data stamp:  2018-08-07T01:20:55Z
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notes: This metadata record was generated by an xslt transformation from a dc metadata record; Transform by Stephen M. Richard, based on a transform by Damian Ulbricht. Run on 2018-08-07T01:20:55Z
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