Abstract:

Gulf killifish were exposed to sublethal serial dilutions of WAF, CWAF, and Corexit that was previously weathered (1, 4, 16 weeks) in experimental mesocosms at salinities of 4, 12, and 18 ppt. After the 96 h exposure period hepatic tissue and whole body gulf killifsh (0.5g) were collected from treatment dilutions for microsomal and cytosolic preparations. Assays were prepared and completed for whole protein concentrations, Cyp1a (ELISA), NaKATPase (ELISA), Ethoxyresorufin-O-deethylase (EROD), and Glutathione S-transferase (GST) activity. Data from these assays and associated protocols are contained within this dataset, which are referred to as Aim 1B within the original proposal.

Suggested Citation:

Chris Green. 2014. Sublethal Bioassays: Killifish response to BP related chemicals across a range of salinities simulating the LA coast. Distributed by: GRIIDC, Harte Research Institute, Texas A&M University–Corpus Christi. doi:10.7266/N7DB7ZS4

Purpose:

This dataset integrates sublethal measures of oil exposure with physiological markers and is linked to sublethal concentrations tested in acute bioassays for Aim 1A.

Data Parameters and Units:

Aim 1B all data summary.xlsx-- 1. EROD GST 18ppt Graphs: EROD (pmol/mL/min), GST (µmol/mL/min) -for Control, Oil, Oil/COREXIT9500 -for Treatment Concentration (%v/v) of 100, 50, 25, 6.25. 2. Summary of cyp1a ELISA: Salinity, Age, Treatment, Dilution, Absolute, Relative 3. EROD GST and Oil Analysis: Salinity, Age, Treatment, Concentration, Replicate, 48hr-Dead, 48hr-Alive, 96hr-Dead, 96hr-Alive, EROD, GST, concentrations of PAHs. 4. EROD GST Graphs: Treatment Dilution (% v/v) verses 1. Fold EROD Induction and 2. Fold GST Induction -for 4PPT, 12PPT, 18PPT -for 1week, 4weeks, 16weeks.

Methods:

GST and EROD data are normalized to protein only, the units are pmol/min/ml (it was a 15 min run). when futher nomalized to controls, it is a fold induction. Water samples were collected from all 18 oil and oil and oil/COREXIT exposed mesocosms as well as from a representative control and COREXIT only mesocosm. Seventy-six PAHs and associated homologues were extracted and analyzed from these water samples by Columbia Analytical Services (Kelso, WA, USA) according to laboratory standard operating procedures. Samples were extracted from 1 L of mesocosm water according to EPA Method 3510C and analyzed by EPA Method 8270D. Reporting limits were typically < 0.03 µg/L, with the exception of the chemically dispersed 18 ppt salinity 1 week weathered samples which were 0.11 µg/L (supplemental data S1). Results of the chemical analyses were used to create “target PAH” and is the sum of all 76 analytes.

Instruments:

Microsomal and cystosolic preparation protein concentrations were determined by the ThermoScientific Coomassie Plus (Bradford) Assay kit (Rockford, IL, USA). Cytosolic GST activities were determined using the Glutathione S-Transferase Assay Kit from Sigma (St. Louis, MO, USA). CYP1A activity was monitored using the ethoxyresorufin-O-deethylase (EROD) assay as described by Willet et al. 1998. Induction of CYP1A was quantified using a method adapted from Scholz et al. 1997. Bio-Tek Synergy HT Multi-Detection Microplate Reater (BioTek Instruments Minooski, VT, USA).

Individual Files: