title: Using silver and bighead carp cell lines for the identification of a unique metabolite fingerprint from thiram-specific chemical exposure.
abstract: Conservation biology often requires the control of invasive species. One method is the development and use of biocides. Identifying new chemicals as part of the biocide registration approval process can require screening millions of compounds. Traditionally, screening new chemicals has been done in vivo using test organisms. Using in vitro (e.g., cell lines) and in silico (e.g., computer models) methods decrease test organism requirements and increase screening speed and efficiency. These methods, however, would be greatly improved by better understanding how individual fish species metabolize selected compounds. We combined cell assays and metabolomics to create a powerful tool to facilitate the identification of new control chemicals. Specifically, we exposed cell lines established from bighead carp and silver carp larvae to thiram (7 concentrations) then completed metabolite profiling to assess the dose-response of the bighead carp and silver carp metabolome to thiram. Forty one of the 700 metabolomic markers identified in bighead carp exhibited a dose-response to thiram exposure compared to silver carp in which 205 of 1590 metabolomic markers exhibited a dose-response. Additionally, we identified 11 statistically significant metabolomic markers based upon volcano plot analysis common between both species. This smaller subset of metabolites formed a thiram-specific metabolomic fingerprint which allowed for the creation of a toxicant specific, rather than a species-specific, metabolomic fingerprint. Metabolomic fingerprints may be used in biocide development and improve our understanding of ecologically significant events, such as mass fish kills.
authors.affiliation: U.S. Geological Survey, La Crosse, WI 54603, USA. Electronic address: jgputnam@usgs.gov.
authors.forename: Joel G
authors.initials: JG
authors.lastname: Putnam
authors.affiliation: U.S. Geological Survey, La Crosse, WI 54603, USA.
authors.forename: Justine E
authors.initials: JE
authors.lastname: Nelson
authors.affiliation: U.S. Fish and Wildlife Service, Onalaska, WI 54650, USA.
authors.forename: Eric M
authors.initials: EM
authors.lastname: Leis
authors.affiliation: U.S. Geological Survey, La Crosse, WI 54603, USA.
authors.forename: Richard A
authors.initials: RA
authors.lastname: Erickson
authors.affiliation: U.S. Geological Survey, La Crosse, WI 54603, USA.
authors.forename: Terrance D
authors.initials: TD
authors.lastname: Hubert
authors.affiliation: U.S. Geological Survey, La Crosse, WI 54603, USA.
authors.forename: Jon J
authors.initials: JJ
authors.lastname: Amberg
mesh_terms: D000818:Animals; D001681:Biological Assay; D002347:Carps; D002460:Cell Line; D003530:Cyprinidae / Q000378:metabolism; D004784:Environmental Monitoring / Q000379:methods*; D005659:Fungicides, Industrial / Q000633:toxicity*; D013045:Species Specificity; D013893:Thiram / Q000633:toxicity*
chemical_list: D005659:Fungicides, Industrial; D013893:Thiram
keywords: Bigheaded carp; Invasive species; Mass spectrometry; Metabolome; Thiram; Toxicity