Abstract:

Three 96 h time course exposures were performed on freshly fertilized mahi-mahi embryos using two types of crude oil (slick (OFS) and source (Mass)) for the purposes of: (1) analyzing whole animal transcriptome responses by RNA-Seq/qPCR analysis, (2) phenotypic anchoring of transcriptional responses to cardiotoxic endpoints by imaging analysis of heart rate and pericardial and yolk sac edema and (3) obtaining additional replicates for qPCR to confirm transcriptional response of select genes identified by RNA-Seq. This dataset contains all of the data associated with the exposures and phenotypic analyses. This dataset also includes level thresholds (e.g., LC25, LC50) for two different weathered states of oil. Tests G001 and G002 were performed with slick (OFS) oil prepared as a high energy water-accommodated fraction (HEWAF). Test G001 failed to produce a dose response due to insufficient mortality; therefore, test G002 was performed as a repeat of test G001 using higher nominal HEWAF concentrations. Test G003 was performed with source (MASS) oil prepared as a HEWAF. Transcriptional response data (i.e. RNA-Seq and qPCR) are provided in a separate dataset (UDI: R4.x258.258:0001).

Suggested Citation:

Edward Mager, Ph.D.. 2016. Time Course Crude Oil Exposures of Early Life Stage Mahi-Mahi for RNA-Seq and Assessment of Cardiotoxicity. Distributed by: GRIIDC, Harte Research Institute, Texas A&M University–Corpus Christi. doi:10.7266/N7K35S21

Publications:

Xu, E. G., Mager, E. M., Grosell, M., Pasparakis, C., Schlenker, L. S., Stieglitz, J. D., … Schlenk, D. (2016). Time- and oil-dependent transcriptomic and physiological responses to Deepwater Horizon oil in mahi-mahi (Coryphaena hippurus) embryos and larvae. Environ. Sci. Technol. doi:10.1021/acs.est.6b02205

Purpose:

Phenotypic anchoring of transcriptional responses observed by RNA-Seq/qPCR to cardiotoxic endpoints by imaging analysis of heart rate and pericardial edema and establishing total PAH effect level thresholds for mortality of early life stage mahi-mahi.

Data Parameters and Units:

Heart rate (beats per minute), pericardial and yolk sac edema (pericardial area in pixels), survival, pH, dissolved oxygen (mg/L), salinity (ppt), ammonia (µM), PAH concentrations (µg/L), Date (DD-MM-YY), period (time period), water temperature (degree C), for video and image files the file name refers to the Tank ID Dilution or Stock Code and replicate (more information can be found in the Tank ID Dilution or Stock Code.csv files) Acute (96 h) Mahi-Mahi Early Life Stage Toxicity Testing: Experiment G001 using slick (OFS) oil prepared as a high energy water-accommodated fraction (HEWAF): Control treatment = 0% oil, Treatment 2 = 0.25% Slick oil, Treatment 3 = 0.5% Slick oil, Treatment 4 = 1% Slick oil, Treatment 5 = 2% Slick oil, Treatment 6 = 4% Slick oil. Experiment G002 using slick (OFS) oil prepared as a high energy water-accommodated fraction (HEWAF): Control treatment = 0% oil, Treatment 2 = 1% Slick oil, Treatment 3 = 2% Slick oil, Treatment 4 = 4% Slick oil, Treatment 5 = 8% Slick oil, Treatment 6 = 16% Slick oil. Experiment G003 using source (MASS) oil prepared as a high energy water-accommodated fraction (HEWAF): Control treatment = 0% oil, Treatment 2 = 0.125% Source oil, Treatment 3 = 0.25% Source oil, Treatment 4 = 0.5% Source oil, Treatment 5 = 1% Source oil, Treatment 6 = 2% Source oil. Four replicates per treatment: Control: C1-C4, Treatment 1: T1-T4, Treatment 2: T5-T8, Treatment 3: T9-T12, Treatment 4: T13-T16, Treatment 5: T17-T20. Ammonia: Standard curve, Rep (replicate), Avg (average), Regression, St Dev (Standard deviation), Conc (concentration). Fluorescence: Sample type, Sample ID, Nominal Dilution (%), Peak Area for Excitation Wavelength 1, Peak Area for Excitation Wavelength 2, Blank Subtracted Sum of Peak Areas 1 and 2, Log Nominal Dilution, Log Blank Subtracted Sum of Peak Areas 1 and 2, Dilution Factor, Measured Dilution (%). PAH Analysis: Units (ug/L), Component, Dilution Factor, Reporting Limit, Detection Limit, Result, Result Notes, Spike Concentration, Percent Recovery, Acceptance Limits, Average, RPD (relative percent difference), Retention Time, Regulatory Limit. Tank ID Dilution or Stock Code: Tank ID Dilution or Stock Code, Start Date, Start time, End date, End time, Nominal Treatment Concentration, Treatment Units, Replicate number. Test Performance Monitoring: Tank ID, Date, Time, Number at test start, Number observed treatment mortalities, Number from non-treatment mortality, Number observed alive. Water Quality Monitoring: Tank ID, Date, Time, Period (Initial, 24h, 48h, 72h, 96h), Water temp. (degrees C), pH, D.O. (mg/L), Salinity (ppt), Total ammonia (uM)

Methods:

Heart rate was determined visually by counting heart beats from videos slowed to 0.5 speed. Pericardial area was measured using ImageJ version 1.46r from a perimeter drawn with the freehand tool enclosing this area

Instruments:

Nikon SMZ800 stereomicroscope with Fire-i400 or Fire-i530c camera, YSI ProODO oxygen meter, PHM201 pH meter, refractometer, Spectramax plate reader, GC/MS-SIM, LS45 fluorescence spectrometer

Individual Files: