Abstract:

This dataset covers GoMRI RFP-II research performed at Louisiana Universities Marine Consortium (LUMCON) during 2016 in Dr. Chesney’s lab. Bay anchovy embryos and 1 day post hatch larvae (1dph) were exposed to HEWAF mixtures at concentrations of 0, 3, 10, 20% HEWAF to evaluate low dose, short duration (2 and 6 hrs.) exposure effects. High-energy water accommodated fraction (HEWAF) were prepared on the day of exposure and diluted with clean seawater to reach the target concentrations. The condition of the embryos and the larvae were assessed for overall survival and condition (malformations) immediately post exposure and then placed in clean seawater, held for 24 hrs. and assessed again 24 hrs. post-exposure.

Suggested Citation:

Chesney, Edward. 2017. Comparison of toxicity of crude oil to bay anchovy (Anchoa mitchilli) embryos and larvae exposed to low doses of HEWAF for short durations. Distributed by: GRIIDC, Harte Research Institute, Texas A&M University–Corpus Christi. doi:10.7266/N7TB157W

Purpose:

This dataset was compiled so the effects of short duration exposure to crude oil with representative Gulf of Mexico fish embryos and larvae could be predicted. Specifically, the study is designed to determine the effects of toxicity of crude oil when embryos and larvae are exposed for durations shorter than typical exposures (>24 hrs) and at low crude oil doses. Establishing the degree of post exposure effects on survival and the rate of developmental malformations was a key element of these studies.

Data Parameters and Units:

Embryo experiment1 filename: BA Embryo_3%, 10%_2hr, 6hr_5-24-16Exp1 Worksheet 1: Exposure&24hpostexposureresults Parameters: Beaker code, Experiment start date, HEWAF concentrations (% of total volume), exposure duration (hrs), replicate number, ML of 25 psu seawater, ML of HEWAF, Number of embryos start, Number of embryos at transfer, Number of dead embryos at transfer, number of unhatched dead embryos, Number hatched healthy larvae, Number hatched dead larvae, Number hatched moribund larvae, Number missing, Notes. Transfers refers to moving embryos to clean seawater. Worksheet 2: Fluorescence Parameters: 3% HEWAF RFUs, 10% HEWAF RFUs. Fluorescence was measured by mixing the appropriate volume of HEWAF (3%=30mL, 10%=100mL) with seawater to make 1L of solution. A sample of this solution was measured three times with a fluorometer to obtain an average fluorescence in RFUs for each concentration. Worksheet 3: GCMS Parameters: sample treatments, 3% BA HEWAF, 10% BA HEWAF, seawater control in ug/L Embryo experiment2 filename: BA Embryo_10%, 20%_2hr, 6hr_5-30-16Exp2 Worksheet 1: Exposure&24hpostexposureresults Parameters: Beaker code, Trial start date, HEWAF concentrations (% of total volume), exposure duration (hrs), replicate number, ML of 25 psu seawater, ML of HEWAF, Number of embryos start, Number of embryos at transfer, Number of dead embryos at transfer, number of unhatched dead embryos, Number hatched healthy larvae, Number hatched dead larvae, Number hatched moribund larvae, Number missing, Ammonia (ppm), Final DO (mg/L), Final Temp. (degrees C), Notes. Transfer refers to moving embyros into 1L clean seawater after exposure. HEWAF concentration is the percentage of the whole 1L volume composed of HEWAF (e.g. 20%=200 mL HEWAF with 800 mL 25 psu seawater to make 1L). Worksheet 2: Malformations Parameters: Tank code, Larval ID, Date, HEWAF concentrations (% of total volume), exposure duration (hrs), Alive (A) or Moribund (M), Malformations: Bent spine, Yolksac, Fin Fold, Eye, Facial, Notes. HEWAF concentration is the percentage of the whole 1L volume composed of HEWAF (e.g. 20%=200 mL HEWAF with 800 mL 25 psu seawater to make 1L). Malformations are given a Y or N to indicate whether or not the given larva exhibits the malformation (Y) or does not (N). Larval ID indicates different larva within a treatment (eg. larva 1 in tank 20%A 6 hr is not the same larva as larva 1 in tank 10%A 2 hr) and only larvae exhibiting some sort of malformation were listed. Worksheet 3: Fluorescence Parameters: 10% HEWAF RFUs, 20% HEWAF RFUs. Fluorescence was measured by mixing the appropriate volume of HEWAF (10%=100mL, 20%=200mL) with seawater to make 1L of solution. A sample of this solution was measured three times with a fluorometer to obtain an average fluorescence in RFUs for each concentration. Worksheet 4: GCMS Parameters: sample treatments, 10% BA HEWAF, 20% BA HEWAF, seawater control in ug/L Embryo experiment3 filename: BA Embryo_10%,20%_2hr, 6hr_6-22-16Exp3 Worksheet 1: Exposure&24hpostexposureresults Parameters: Beaker code, Trial date, HEWAF concentrations (% of total volume), exposure duration (hrs), replicate number, ML of 25 psu seawater, ML of HEWAF, Number of embryos start, Number of embryos at transfer, Number of dead embryos at transfer, number of unhatched dead embryos, Number hatched healthy larvae, Number hatched dead larvae, Number hatched moribund larvae, Number missing, Notes. Transfer refers to moving embryos into 1L of clean seawater after exposure. HEWAF concentration is the percentage of the whole 1L volume composed of HEWAF (e.g. 20%=200 mL HEWAF with 800 mL 25 psu seawater to make 1L). Worksheet 2: Malformations Parameters: Tank code, Larval ID, Date, HEWAF concentrations (% of total volume), exposure duration (hrs), Alive (A) or Moribund (M), Malformations: Bent spine, Yolksac, Fin Fold, Eye, Facial, Notes. HEWAF concentration is the percentage of the whole 1L volume composed of HEWAF (e.g. 20%=200 mL HEWAF with 800 mL 25 psu seawater to make 1L). Malformations are given a Y or N to indicate whether or not the given larva exhibits the malformation (Y) or does not (N). Larval ID indicates different larva within a treatment (eg. larva 1 in tank 20%A 6 hr is not the same larva as larva 1 in tank 10%A 2 hr) and only larvae exhibiting some sort of malformation were listed. Worksheet 3: Fluorescence Parameters: 10% HEWAF RFUs, 20% HEWAF RFUs. Fluorescence was measured by mixing the appropriate volume of HEWAF (10%=100mL, 20%=200mL) with seawater to make 1L of solution. A sample of this solution was measured three times with a fluorometer to obtain an average fluorescence in RFUs for each concentration. Worksheet 4: GCMS Parameters: sample treatments, seawater control, Ba 3% HEWAF 2 and 6 hr embryo, Ba 10% HEWAF 2 and 6 hr embryo in ug/L Larvae experiment1 filename: BAlarvae3%10%2,6hrHEWAFexposures7-21-2016exp1 Worksheet 1: Exposure conditions Parameters: Beaker code, start date, start time, Nominal concentration (% HEWAF), replicate number, larvae per beaker, exposure duration (hrs), ML of 25 psu seawater, ML of HEWAF, Notes. HEWAF concentration is the percentage of the whole 1L volume composed of HEWAF (e.g. 20%=200 mL HEWAF with 800 mL 25 psu seawater to make 1L). 60 bay anchovy embryos were placed in each treatment beaker and left overnight to hatch. After hatching, the larvae were exposed to the desired oil concentration for the given amount of time. Worksheet 2: Assessment (Immed. post-exp) Parameters: Beaker code, Assessment Date, Assessment time, number unhatched dead embryos, number hatched healthy larvae, number of hatched dead larvae, number of moribund larvae, notes Worksheet 3: PostexposureMalformations Parameters: Beaker code, larval ID, date, HEWAF concentration (%), treatment time (hours), Vitality: Alive (A) or moribund (M), Malformation categories: bent spine, yolksac, finfold, eye, facial, Notes. Malformations are given a Y or N to indicate whether or not the given larva exhibits the malformation (Y) or does not (N). Worksheet 4: 24 hr postexposureassessment Parameters: Beaker code, Date, Time, number of larvae transferred to growout beakers, number healthy larvae, number dead larvae, number moribund larvae. 60 larvae were exposed of which 25 were moved to clean seawater. About 35 larvae were assessed immediately post-exposure, while the other 25 were assessed 24 hours later. Worksheet 5: 24 h. postexposuremalformations Parameters: Beaker code, Larval ID, Date, HEWAF concentration (%), treatment time (hours), Vitality: Alive (A) or moribund (M), Malformation categories: bent spine, yolksac, finfold, eye, facial, Notes Worksheet 6: HEWAFFluorescence Parameters: 3% HEWAF, 10% HEWAF Worksheet 7: GCMS Parameters: sample treatments, seawater control, BA 3% 2 and 6 hrs HEWAF embryo, BA 10% 2 and 6 hrs HEWAF embryo Larvae experiment2 filename: BA Larvae_10%_2hr,6hrHEWAFexposures6-7-16exp2 Worksheet 1: Exposure conditions Parameters: Beaker code, trial start date, replicate number, larvae per beaker, exposure duration (hrs), HEWAF concentrations (% of total volume), ML of 25 psu seawater, ML of HEWAF, Ammonia (ppm), Notes. HEWAF concentration is the percentage of the whole 1L volume composed of HEWAF (eg. 20%= 200 mL HEWAF with 800 mL seawater to make 1L). 60 embryos were placed in each treatment beaker and left overnight to hatch. After hatching, the larvae were exposed to the target oil concentration for the given amount of time (2 or 6 hrs) and then condition assessed. Worksheet 2: Assessment (Immed. post-exp) Parameters: Beaker code, Assessment Date, Assessment time, number unhatched dead embryos, number healthy larvae, number of dead larvae, number of moribund larvae, number of missing larvae, notes. 25 Larvae were dipped out of treatment water and placed in clean water which was then added to beakers where the larvae were allowed to grow out for 24 hours. See 24 h post-exposureassessment tab. The rest of the larvae were assessed for condition and discarded. Worksheet 3: Malformations (immed. post exp.) Parameters: HEWAF concentration (%), Exposure duration (hrs), Alive (A) or moribund (M), Malformation categories: bent spine, yolksac, finfold, eye, facial, Notes. HEWAF concentration is the percentage of the whole 1L volume composed of HEWAF (eg. 20%= 200 mL HEWAF with 800 mL seawater to make 1L). Malformations are giving a Y or N to indicate whether or not the given larva exhibits (Y) the malformation or does not (N). Larval ID is meant to indicate a specific larva within a treatment (eg. larva 1 in tank 20%A 6 hr is not the same larva as larva 1 in tank 10%A 2 hr). Note that only those larvae that exhibited some sort of malformation were listed. Worksheet 4: 24 hr postexposureassessment Parameters: Beaker code, Date, Time, number of larvae transferred to growout beakers, number healthy larvae, number dead larvae, number moribund larvae, number missing larvae Worksheet 5: 24 h. postexposuremalformations Parameters: Beaker code, Larval ID, Date, HEWAF concentration (%), Exposure duration (hrs), Alive (A) or moribund (M), Malformation categories: bent spine, yolksac, finfold, eye, facial, Notes. HEWAF concentration is the percentage of the whole 1L volume composed of HEWAF (eg. 20%= 200 mL HEWAF with 800 mL seawater to make 1L). Malformations are giving a Y or N to indicate whether or not the given larva exhibits (Y) the malformation or does not (N). Larval ID is meant to indicate a specific larva within a treatment (eg. larva 1 in tank 20%A 6 hr is not the same larva as larva 1 in tank 10%A 2 hr). Note that only those larvae that exhibited some sort of malformation were listed. Worksheet 6: HEWAFFluorescence Parameters: 10% HEWAF (RFUs) Worksheet 7: GCMS Parameters: sample treatments, seawater control, 10% BA HEWAF in ug/L

Methods:

Exposure experiments (bay anchovy embryos and 1 DPH larvae): Bay anchovy larvae or embryos, were exposed to HEWAF at various concentrations for short durations (2 or 6 h exposures). Once the exposure time period was complete the number and condition of the larvae or embryos were noted. After exposures all living embryos and a subset (25 apparently healthy larvae) were moved to clean seawater and held for 24 hrs. After 24 hrs. the survival and condition of remaining larvae in all treatment was evaluated. Petroleum analyte levels were estimated by GCMS.

Instruments:

Salinity was measured using a VitalSine ™ SR-6 Salinity Refractometer. Dissolved oxygen was measured using a YSI Professional Optical Dissolved Oxygen (ProODO™) Instrument. Ammonia levels were measured using an API Aquarium Pharmaceuticals Ammonia Test Kit. Relative fluorescent units (RFUs) were measured using a Trilogy Laboratory Fluorometer, model # 7200-000.

Individual Files: