Toxicology across scales: Cell population growth in vitro predicts reduced fish growth

Cells in vitro can save lives of experimental fish used in chemical safety testing. Environmental risk assessment of chemicals is essential but often relies on ethically controversial and expensive methods. We show that tests using cell cultures, combined with modeling of toxicological effects, can replace tests with juvenile fish. Hundreds of thousands of fish at this developmental stage are annually used to assess the influence of chemicals on growth. Juveniles are more sensitive than adult fish, and their growth can affect their chances to survive and reproduce. Thus, to reduce the number of fish used for such tests, we propose a method that can quantitatively predict chemical impact on fish growth based on in vitro data. Our model predicts reduced fish growth in two fish species in excellent agreement with measured in vivo data of two pesticides. This promising step toward alternatives to fish toxicity testing is simple, inexpensive, and fast and only requires in vitro data for model calibration.

[1]  I. Wilson,et al.  Investigation of the Alamar Blue (resazurin) fluorescent dye for the assessment of mammalian cell cytotoxicity. , 2000, European journal of biochemistry.

[2]  S. J. Caldwell,et al.  Development and characterization of a rainbow trout liver cell line expressing cytochrome P450-dependent monooxygenase activity , 1993, Cell Biology and Toxicology.

[3]  Vivian R. Dayeh,et al.  Applications and potential uses of fish gill cell lines: examples with RTgill-W1 , 2009, In Vitro Cellular & Developmental Biology - Animal.

[4]  Vivian R. Dayeh,et al.  Chapter 2 Use of fish cell lines in the toxicology and ecotoxicology of fish. Piscine cell lines in environmental toxicology , 2005 .

[5]  G. Ankley,et al.  Propiconazole inhibits steroidogenesis and reproduction in the fathead minnow (Pimephales promelas). , 2013, Toxicological sciences : an official journal of the Society of Toxicology.

[6]  Roman Ashauer,et al.  General unified threshold model of survival--a toxicokinetic-toxicodynamic framework for ecotoxicology. , 2011, Environmental science & technology.

[7]  P. Taylor,et al.  Von Bertalanffy's Growth Equation Should Not Be Used to Model Age and Size at Maturity , 1997, The American Naturalist.

[8]  I. Conlon,et al.  Size Control in Animal Development , 1999, Cell.

[9]  S. J. Caldwell,et al.  Development of a cell line from primary cultures of rainbow trout, Oncorhynchus mykiss (Walbaum), gills , 1994 .

[10]  Trevor Hastie,et al.  Regularization Paths for Generalized Linear Models via Coordinate Descent. , 2010, Journal of statistical software.

[11]  J. Nichols,et al.  Physiologically based toxicokinetic modeling of three waterborne chloroethanes in rainbow trout (Oncorhynchus mykiss). , 1991, Toxicology and applied pharmacology.

[12]  A. Fisk,et al.  Bioaccumulation and biotransformation of chiral triazole fungicides in rainbow trout (Oncorhynchus mykiss). , 2006, Aquatic toxicology.

[13]  Kristin Schirmer,et al.  Development of a partition-controlled dosing system for cell assays. , 2010, Chemical research in toxicology.

[14]  H. Zou,et al.  Regularization and variable selection via the elastic net , 2005 .

[15]  Holger Schüttrumpf,et al.  Physiologically-based toxicokinetic models help identifying the key factors affecting contaminant uptake during flood events. , 2014, Aquatic toxicology.

[16]  E D Kroese,et al.  REACH, non-testing approaches and the urgent need for a change in mind set. , 2009, Regulatory toxicology and pharmacology : RTP.

[17]  Roberto Maass-Moreno,et al.  Fitting Models to Biological Data Using Linear and Nonlinear Regression: A Practical Guide to Curve Fitting.ByHarvey Motulskyand, Arthur Christopoulos.Oxford and New York: Oxford University Press. $65.00 (hardcover); $29.95 (paper). 351 p; ill.; index. ISBN: 0–19–517179–9 (hc); 0–19–517180–2 (pb). 2 , 2005 .

[18]  Roman Ashauer,et al.  Predicting Concentrations of Organic Chemicals in Fish by Using Toxicokinetic Models , 2012, Environmental science & technology.

[19]  J. Devillers,et al.  A physiologically based toxicokinetic model for the zebrafish Danio rerio. , 2014, Environmental science & technology.

[20]  L. Bertalanffy,et al.  A quantitative theory of organic growth , 1938 .

[21]  C. Cowan-Ellsberry,et al.  Toward improved models for predicting bioconcentration of well‐metabolized compounds by rainbow trout using measured rates of in vitro intrinsic clearance , 2013, Environmental toxicology and chemistry.

[22]  T. Hutchinson,et al.  Analysis of the ECETOC aquatic toxicity (EAT) database III - comparative toxicity of chemical substances to different life stages of aquatic organisms , 1998 .

[23]  K. Schirmer.  Proposal to improve vertebrate cell cultures to establish them as substitutes for the regulatory testing of chemicals and effluents using fish. , 2006, Toxicology.

[24]  J. Mckim,et al.  A physiologically based toxicokinetic model for lake trout (Salvelinus namaycush). , 2001, Aquatic toxicology.

[25]  J. Nichols,et al.  Physiologically-based toxicokinetic modeling of three waterborne chloroethanes in channel catfish, Ictalurus punctatus , 1993 .

[26]  Vivian R. Dayeh,et al.  Applying whole-water samples directly to fish cell cultures in order to evaluate the toxicity of industrial effluent. , 2002, Water research.

[27]  M. Gravell,et al.  A PERMANENT CELL LINE FROM THE FATHEAD MINNOW (PIMEPHALES PROMELAS) * , 1965, Annals of the New York Academy of Sciences.

[28]  J. Hermens,et al.  Modes of action in ecotoxicology: their role in body burdens, species sensitivity, QSARs, and mixture effects. , 2002, Environmental science & technology.

[29]  Karline Soetaert,et al.  Solving Differential Equations in R: Package deSolve , 2010 .

[30]  Theo Vermeire,et al.  Risk assessment of chemicals , 2021, Bioanalytical Tools in Water Quality Assessment.

[31]  K. Schirmer,et al.  The Use of Fish‐Derived Cell Lines for Investigation of Environmental Contaminants , 2003, Current Protocols in Toxicology.

[32]  Hilda Witters,et al.  A European perspective on alternatives to animal testing for environmental hazard identification and risk assessment. , 2013, Regulatory toxicology and pharmacology : RTP.

[33]  M. Hooten,et al.  The influence of external subsidies on diet, growth and Hg concentrations of freshwater sport fish: implications for management and fish consumption advisories , 2012, Ecotoxicology.

[34]  Roman Ashauer,et al.  Measured and Modeled Toxicokinetics in Cultured Fish Cells and Application to In Vitro - In Vivo Toxicity Extrapolation , 2014, PloS one.

[35]  Karline Soetaert,et al.  Inverse Modelling, Sensitivity and Monte Carlo Analysis in R Using Package FME , 2010 .

[36]  M. Andersen,et al.  A physiologically based toxicokinetic model for the uptake and disposition of waterborne organic chemicals in fish. , 1990, Toxicology and applied pharmacology.

[37]  J. B. Sprague.  Measurement of pollutant toxicity to fish—III: Sublethal effects and “safe” concentrations , 1971 .

[38]  Kristin Schirmer,et al.  Predicting fish acute toxicity using a fish gill cell line-based toxicity assay. , 2013, Environmental science & technology.

[39]  Roman Ashauer,et al.  Toxicokinetic-toxicodynamic modelling of survival of Gammarus pulex in multiple pulse exposures to propiconazole: model assumptions, calibration data requirements and predictive power , 2012, Ecotoxicology.

[40]  J. Tietge,et al.  Physiologically based toxicokinetic model for maternal transfer of 2,3,7,8‐tetrachlorodibenzo‐p‐dioxin in brook trout (Salvelinus fontinalis) , 1998 .

[41]  M. Maines.  Current protocols in toxicology , 1999 .

[42]  D. Dixon,et al.  Methodology for demonstrating and measuring the photocytotoxicity of fluoranthene to fish cells in culture. , 1997, Toxicology in vitro : an international journal published in association with BIBRA.

[43]  Stefan Scholz,et al.  Adverse outcome pathways during early fish development: a conceptual framework for identification of chemical screening and prioritization strategies. , 2011, Toxicological sciences : an official journal of the Society of Toxicology.

[44]  S. Nesnow,et al.  Cytotoxic effects of propiconazole and its metabolites in mouse and human hepatoma cells and primary mouse hepatocytes. , 2008, Toxicology in vitro : an international journal published in association with BIBRA.

[45]  A. Christopoulos,et al.  Fitting Models to Biological Data Using Linear and Nonlinear Regression: A Practical Guide to Curve Fitting , 2004 .