Abstract:
This dataset contains the raw data obtained in support of the publication: Dasgupta, S., DiGiulio, R.T., Drollette, B.D., L Plata, D., Brownawell, B.J., & McElroy, A.E. 2016. Hypoxia depresses CYP1A induction and enhances DNA damage, but has minimal effects on antioxidant responses in sheepshead minnow (Cyprinodon variegatus) larvae exposed to dispersed crude oil. Aquat. Toxicol. 177 (250-260). (WAF) or chemically enhanced dispersed crude oil (CEWAF) were made from Southern Louisiana Sweet Crude (Surrogate) Oil. CYP1A activity was measured in single live larvae using the in vivo EROD (ethoxy resorufin O-deethylase) assay. DNA damage was assessed using the Comet Assay on whole larvae. Activities of a suite of oxidative stress enzymes including catalase (CAT), glutathione reductase (GR), glutathione peroxidase (GPx), glutathione-S-transferase (GST), superoxide dismutase (SOD), and glutamate cysteine ligase (GCL) were measured, as well as levels of total, reduced and oxidized glutathione (tGSH, GSH, GSSG), and lipid peroxidation, using the thiobarbituric-acid reactive substances (TBARS) assay.
Suggested Citation:
Subham Dasgupta. 2017. Data on hypoxia effects on cellular responses to WAF and CEWAF solutions in sheepshead minnow larvae. Distributed by: GRIIDC, Harte Research Institute, Texas A&M University–Corpus Christi. doi:10.7266/N7WM1BSH
Data Parameters and Units:
EROD activity nM of resorufin (RF) excreted per hr per larvae. All other enzyme activities reported as nM/min-mg protein. Total and reduced and oxidized glutathione nM/mg larvae. TBARS nM of malondialdehyde (MDA)/mg protein. Comet assay data is in Olive Tail Moment reported as (OTM). Data on total aromatic hydrocarbons (AH) in mg/L. Data on individual aromatic hydrocarbons ng/mL. Oil data is in ng/g. Total Gluthathione (TGSH) in nm/g wet weight. Reduced Glutathione (GSH) in nm/g wet weight. Oxidized Glutathione (GSSG) in nM/g wet weight. Compound, ASW (ng/mL), WAF (ng/mL), CEWAF (ng/mL), Macondo Oil (ng/g), Detection limit (ppb), n.d = not detected, Experiment, Solution, Nominal loading (mg/L), TPH (mg/L)
Methods:
WAF: CYP1A activity in sheepshead minnow larvae after exposure to WAF solutions for 24 hrs as assessed using ethoxyresorufin )-deethylase (EROD) assay on live larvae. 10 larvae per treatment. Resorufin (RF) production measured as proxy for CYP1A activity. Larvae were incubated with dosing solution+ EROD mixture and RF production measured after 24 hrs spectrofluorometrically. CEWAF: CYP1A activity in sheepshead minnow larvae after exposure to CEWAF solutions for 24 hrs as assessed using ethoxyresorufin )-deethylase (EROD) assay on live larvae. 10 larvae per treatment. Resorufin (RF) production measured as proxy for CYP1A activity. Larvae were incubated with dosing solution+ EROD mixture and RF production measured after 24 hrs spectrofluorometrically. WAF-CEWAF: CYP1A activity in sheepshead minnow larvae after exposure to WAF/CEWAF solutions under normoxic (~21% O2) or hypoxic (~5% O2) conditions for 24 hrs as assessed using ethoxyresorufin-deethylase (EROD) assay on live larvae. 10 larvae per treatment. Resorufin (RF) production measured as proxy for CYP1A activity. Larvae were incubated with dosing solution+ EROD mixture and RF production measured after 24 hrs spectrofluorometrically. GST: Gluathione -S-Transferase Activity (GST) Data represents enzyme activity from 2 experiments. Each experiment had 3 replicates per treatment and 24 larvae per replicate. Under normoxic (~20% O2) and hypoxic (~5% O2) levels. CEWAF AH= ~150 mg/L, nominal loading = 600 mg/L Glutathione Peroxidase Activity (GPx): Data represents enzyme activity from 2 experiments. Each experiment had 3 replicates per treatment and 24 larvae per replicate. Under normoxic (~20% O2) and hypoxic (~5% O2) levels. CEWAF AH= ~150 mg/L, nominal loading = 600 mg/L Glutathione Reductase Activity (GR): Data represents enzyme activity from 2 experiments. Each experiment had 3 replicates per treatment and 24 larvae per replicate. Under normoxic (~20% O2) and hypoxic (~5% O2) levels. CEWAF AH= ~150 mg/L, nominal loading = 600 mg/L Catalase Activity (CAT): Data represents enzyme activity from 2 experiments. Each experiment had 3 replicates per treatment and 24 larvae per replicate. Under normoxic (~20% O2) and hypoxic (~5% O2) levels. CEWAF AH= ~150 mg/L, nominal loading = 600 mg/L Superoxide Dismutase Activity (SOD): Data represents enzyme activity from 2experiments. Each experiment had 3 replicates per treatment and 24 larvae per replicate. Under normoxic (~20% O2) and hypoxic (~5% O2) levels. CEWAF AH= ~150 mg/L, nominal loading = 600 mg/L Glutamate Cysteine Ligase Activity (GCL): Data represents enzyme activity from 2 experiments. Each experiment had 3 replicates per treatment and 24 larvae per replicate. Under normoxic (~20% O2) and hypoxic (~5% O2) levels. CEWAF AH= ~150 mg/L, nominal loading = 600 mg/L Total Glutathione (TGSH): Data represents TGSH activity from 1 experiment with 3 replicates per treatment and 10 larvae per replicate. Under normoxic (~20% O2) and hypoxic (~5% O2) levels. CEWAF AH= ~150 mg/L, nominal loading = 600 mg/L. Normalized to weight of larvae. Reduced Glutathione (GSH): Data represents GSH activity from 1 experiment with 3 replicates per treatment and 10 larvae per replicate. Under normoxic (~20% O2) and hypoxic (~5% O2) levels. CEWAF AH= ~150 mg/L, nominal loading = 600 mg/L. Normalized to weight of larvae. Oxidized Glutathione (GSSG): Data represents GSSG activity from 1 experiment with 3 replicates per treatment and 10 larvae per replicate. Under normoxic (~20% O2) and hypoxic (~5% O2) levels. CEWAF AH= ~150 mg/L, nominal loading = 600 mg/L. Normalized to weight of larvae. TBARS is a measure of lipid peroxidation: Data represents TBARS activity from 2 experiments. Each experiment had 3 replicates per treatment and 24 larvae per replicate. Both under normoxic (~20% O2) and hypoxic (~5% O2) levels. CEWAF AH= ~150 mg/L, nominal loading = 600 mg/L. Normalized to weight of larvae. Olive Tail Moment (OTM) as a measure of DNA damage: Data represents enzyme activity from 2 experiments. Each experiment had 3 replicates per treatment and 10 larvae per replicate. Under normoxic (~20% O2) and hypoxic (~5% O2) levels. WAF AH ~10 mg/L, CEWAF AH= ~150 mg/L, nominal loading = 600 mg/L. Total petroleum hydrocarbons were measured in all Water Accommodated Fractions (WAFs) and Chemically Enhanced Water Accommodated Fractions (CEWAFs) evaluated in these experiments, using fluorescence spectroscopy. Total Petroleum Hydrocarbons (TPH) were estimated from standard curve prepared from different dilutions of oil dissolved in 50:50 Isopropanol:Seawater.
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