Research ArticleECOTOXICOLOGY

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

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Science Advances  07 Aug 2015:
Vol. 1, no. 7, e1500302
DOI: 10.1126/sciadv.1500302
  • Fig. 1 Time-dependent chemical concentration-response curves.

    (A and B) Results for cyproconazole (A) and for propiconazole (B). Symbols, measured values; solid lines, values described by sigmoidal concentration-response curves. Data points for 120-hour exposure (dashed lines) were predicted by the GUTS-SD model, and the sigmoidal concentration-response curve was fitted to these values. CIs are given in table S8.

  • Fig. 2 Interpolation of the predicted fish weight reduction to other chemical concentrations.

    Symbols represent predictions with model uncertainty (Materials and Methods: Linking cell population growth to whole-organism growth) obtained on the basis of measured in vitro data and the von Bertalanffy growth model, and lines represent fitted model with 95% CIs (Materials and Methods: Interpolation to other chemical concentrations).

  • Fig. 3 Measured reduction of fish weight [“●” (each dot represents one fish); mean, “−”] and predictions including model uncertainty (Materials and Methods: Linking cell population growth to whole organism growth) based on in vitro cell population growth data (“Embedded Image”) for different chemical concentrations.

    (A and B) Results for cyproconazole (A) and for propiconazole (B). Weight reduction is presented as percent of weight of control samples after 62-day exposure for cyproconazole and 31-day exposure for propiconazole. We assumed that the total mass of all fish cells is the same as fish weight. For cyproconazole, all tested concentrations caused a significant effect on fish weight, whereas for propiconazole, the three highest concentrations caused significant effect on fish weight [P < 0.05, analysis of variance (ANOVA) and Dunnett’s multiple comparison test, performed in GraphPad Prism].

  • Table 1 Chemical concentrations in water and medium, their corresponding predicted internal concentrations in fish gills (in vivo) and gill cells (in vitro), and their respective effects on fish and fish cell weight.
    Concentration
    in water
    (mg/liter)
    in FELS
    Concentration
    in fish gills
    and gill cells
    (μmol/g)
    Concentration
    in medium
    (mg/liter)
    in cell
    proliferation
    experiments
    Fish
    weight*
    (% of
    control)
    Modeled
    cell
    weight
    (% of
    control)
    Cyproconazole (log KOW = 2.9; molecular weight, 291.78 g/mol)
    2.40.2961.5All dead47.9 (±9.62)
    1.20.1480.7551.2 (±23.1)53.1 (±11.1)
    0.60.0740.37570.9 (±15.6)66.4 (±15.9)
    0.30.0370.187574.3 (±13.4)74.1 (±13.2)
    0.150.0190.0937578.2 (±12.5)85.9 (±14.2)
    Propiconazole (log KOW = 3.72; molecular weight, 342.22 g/mol)
    1.00.4032.323.2 (±13.2)29.6 (±5.67)
    0.50.2021.1574.1 (±22.3)60.5 (±8.62)
    0.250.1010.57583.2 (±20.4)77.5 (±13.0)
    0.1250.0500.287593.9 (±20.7)87.9 (±11.3)
    0.06250.0250.1437596.1 (±24.6)93.6 (±11.8)

    *Measured in FELS studies at day 62 for cyproconazole (rainbow trout) and at day 31 for propiconazole (fathead minnow).

    †Predicted with von Bertalanffy growth model on the basis of in vitro cell proliferation experiments (5-day exposure).

    Supplementary Materials

    • Supplementary material for this article is available at http://advances.sciencemag.org/cgi/content/full/1/7/e1500302/DC1

      Table S1. Physicochemical properties of cyproconazole and propiconazole.

      Table S2. Measured concentrations of cyproconazole in exposure medium during the cell survival experiment (±SD): experiment 1, three technical replicates.

      Table S3. Measured concentrations of cyproconazole in exposure medium during the cell survival experiment (±SD): experiment 2, three technical replicates.

      Table S4. Measured concentrations of propiconazole in exposure medium during the cell survival experiment (±SD): experiment 1, three technical replicates.

      Table S5. Measured concentrations of propiconazole in exposure medium during the cell survival experiment (±SD): experiment 2, three technical replicates.

      Table S6. Measured concentrations of cyproconazole in exposure medium during cell proliferation experiments (0±SD) with three technical replicates per experiment.

      Table S7. Measured concentrations of propiconazole in exposure medium during cell proliferation experiments (±SD) with three technical replicates per experiment.

      Table S8. Parameters of the Hill slope equations for different time points (49).

      Table S9. Estimates of GUTS-SD parameters for cell survival (in vitro) (lower/upper 95% confidence limit) (21, 50).

      Table S10. Estimated parameters of the von Bertalanffy growth model.

      Fig. S1. Survival fraction of RTgill-W1 cells exposed to a range of concentrations of the two pesticides over time.

      Fig. S2. Proliferation of RTgill-W1 cells exposed to different cyproconazole concentrations (left column) and predicted reduction of fish weight caused by the respective concentration (right column: dashed lines represent the model uncertainty).

      Fig. S3. Proliferation of RTgill-W1 cells exposed to different propiconazole concentrations (left column) and predicted reduction of fish weight caused by the respective concentration (right column: dashed lines represent the model uncertainty).

    • Supplementary Materials

      This PDF file includes:

      • Table S1. Physicochemical properties of cyproconazole and propiconazole.
      • Table S2. Measured concentrations of cyproconazole in exposure medium during the cell survival experiment (±SD): experiment 1, three technical replicates.
      • Table S3. Measured concentrations of cyproconazole in exposure medium during the cell survival experiment (±SD): experiment 2, three technical replicates.
      • Table S4. Measured concentrations of propiconazole in exposure medium during the cell survival experiment (±SD): experiment 1, three technical replicates.
      • Table S5. Measured concentrations of propiconazole in exposure medium during the cell survival experiment (±SD): experiment 2, three technical replicates.
      • Table S6. Measured concentrations of cyproconazole in exposure medium during cell proliferation experiments (0±SD) with three technical replicates per experiment.
      • Table S7. Measured concentrations of propiconazole in exposure medium during cell proliferation experiments (±SD) with three technical replicates per experiment.
      • Table S8. Parameters of the Hill slope equations for different time points (49).
      • Table S9. Estimates of GUTS-SD parameters for cell survival (in vitro) (lower/upper 95% confidence limit) (21, 50).
      • Table S10. Estimated parameters of the von Bertalanffy growth model.
      • Fig. S1. Survival fraction of RTgill-W1 cells exposed to a range of concentrations of the two pesticides over time.
      • Fig. S2. Proliferation of RTgill-W1 cells exposed to different cyproconazole concentrations (left column) and predicted reduction of fish weight caused by the respective concentration (right column: dashed lines represent the model uncertainty).
      • Fig. S3. Proliferation of RTgill-W1 cells exposed to different propiconazole concentrations (left column) and predicted reduction of fish weight caused by the respective concentration (right column: dashed lines represent the model uncertainty).

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