Abstract:

As part of an ongoing assessment of the long-term impacts of oil and dispersants on ecosystem health experiments were performed to examine the effects of exposure on different species of fish and varying routes of exposure. As a part of this study we directly assessed the mutagenic potential of the contaminated treatments as well as the surrounding environment in which the fish were contained. For these assessments we used the biologically based assay of DNA damage termed the Microscreen Assay. General microbiological parameters including total viral and bacterial counts and culturable bacterial counts were also measured. Fish experiment data are available in GRIIDC datasets R4.x267.000:0040 and R4.x267.000:0041.

Suggested Citation:

McDaniel, Lauren. 2018. Fish Exposure Studies: Associated water, sediment and food microbiology and mutagenic potential. Distributed by: GRIIDC, Harte Research Institute, Texas A&M University–Corpus Christi. doi:10.7266/N7Z60MFF

Purpose:

To directly measure the mutagenic potential of the oil contaminated treatments utilized in fish exposure experiments. Additionally, the mutagenicity of the surrounding media was assessed to ensure the exposures were contained within the desired treatment milieu. For example, when the oil exposure was contaminated the surrounding water was assessed for mutagenicity to ensure the oil didn’t significantly leach out of the food. Similar assessments were performed for water column and sediment exposures. Basic microbiologic parameters were also be measured to ensure comparable environments between treatment and control exposures.

Data Parameters and Units:

Fish Exposure Samples – PAH: Sample ID, Sample Date (MM/DD/YYYY), Sample type, Trial number/type, Microscreen % Change, Microscreen pos/neg, Naphthalene (ng/L), C1N (ng/L), C2N (ng/L), C3N (ng/L), C4N (ng/L), Acenaphthylene (ng/L), Acenaphthene (ng/L), Fluorene (ng/L), C1F (ng/L), C2F (ng/L), C3F (ng/L), Dibenzothiophene (ng/L), C1D (ng/L), C2D (ng/L), C3D (ng/L), C4D (ng/L), Phenanthrene (ng/L), Anthracene (ng/L), C1PH/A (ng/L), C2PH/A (ng/L), C3PH/A (ng/L), C4PH/A (ng/L), Fluoranthene (ng/L), Pyrene (ng/L), C1F/Py (ng/L), C2F/Py (ng/L), C3F/Py (ng/L), C4F/Py (ng/L), Benzo[B]fluorine (ng/L), Napthobenzothiophene (ng/L), C1NBT (ng/L), C2NBT (ng/L), C3NBT (ng/L), C4NBT (ng/L), Benzo[A]anthracene (ng/L), Chrysene (ng/L), C1BA/C (ng/L), C2BA/C (ng/L), C3BA/C (ng/L), C4BA/C (ng/L), Benzo[B]fluoranthene (ng/L), Benzo[K]fluoranthene (ng/L), Benzo[E]pyrene (ng/L), Benzo[A]pyrene (ng/L), Perylene (ng/L), Indeno[1,2,3,-cd]pyrene (ng/L), Dibenzo[a,h]anthracene (ng/L), Benzo[g,h,i]perylene (ng/L), Total (ng/L); Trial 103 Food Exposure: 11/15 Microscreen Mutagenicity Assay of Fish Food from Exposure Study. Assay Performed 4/6/16 –Sample (Positive control (MitC), Negative control (FSW), Control Fish Food, Low Oil Fish Food, High Oil Fish Food), Sample ID (null), Replicate (1-1, 1-2, 2-1, 2-2), Dilution, Number of Plaques, Volume of sample (usually for 100 ul), Plate Factor, pfu/ml Average, StDev, t-test; 11/15 Plate counts of tank bacterial abundances from Fish Food Exposure Trial 103. Mote Red Drum Dietary Oil Exposure Experiment. Sampling Date 11/24/15 (note, sampled immediately post feeding) – Treatment Sample Name (Control - Sample 1, Treatment - Low oil exposure, Treatment - High oil exposure), Sample ID (null), Replicate (1-1, 1-2, 2-1, 2-2, 3-1, 3-2), Dilution Factor, Plate Factor, Colonies, Vol factor 50 ul plated, CFUs/ml, Average, StdDev; Mote Red Drum Dietary Oil Exposure Experiment. Sampling Date 11/27/15 (note, sampled prior to feeding) – Treatment Sample Name (Control - Sample 1, Treatment - Low oil exposure, Treatment - High oil exposure), Sample ID (null), Replicate (1-1, 1-2, 2-1, 2-2, 3-1, 3-2), Dilution Factor, Plate Factor, Colonies, Vol factor 50 ul plated , CFUs/ml, Average, StdDev; Mote Red Drum Dietary Oil Exposure Experiment. Sampling Date 12/01/15 (note, sampled prior to feeding) – Treatment Sample Name (Control - Sample 1, Treatment - Low oil exposure, Treatment - High oil exposure), Sample ID (null), Replicate (1-1, 1-2, 2-1, 2-2, 3-1, 3-2), Dilution Factor, Plate Factor, Colonies, Vol factor 50 ul plated, CFUs/ml, Average, StdDev; Trial 103 Food Exposure 11-16: 11/16 Plate counts of tank bacterial abundances from Fish Food Exposure Trial 103. Mote Red Drum Dietary Oil Exposure Experiment. Sampling Date 11/3/16 (note, sampled immediately post feeding) – Treatment Sample Name (Control - Sample 1, Treatment - Low oil exposure, Treatment - High oil exposure), Sample ID (null), Replicate (1-1, 1-2, 2-1, 2-2, 3-1, 3-2), Dilution Factor, Plate Factor, Colonies, Vol factor 50 ul plated, CFUs/ml, Average, StdDev; Mote Red Drum Dietary Oil Exposure Experiment. Sampling Date 11/10/16 (note, sampled after feeding) – Treatment Sample Name (Control - Sample 1, Treatment - Low oil exposure, Treatment - High oil exposure), Sample ID (null), Replicate (1-1, 1-2, 2-1, 2-2, 3-1, 3-2), Dilution Factor, Plate Factor, Colonies, Vol factor 50 ul plated, CFUs/ml, Average, StdDev; Mote Red Drum Dietary Oil Exposure Experiment. Sampling Date 11/16/16 (note, sampled after feeding) – Treatment Sample Name (Control - Sample 1, Treatment - Low oil exposure, Treatment - High oil exposure), Sample ID (null), Replicate (1-1, 1-2, 2-1, 2-2, 3-1, 3-2), Dilution Factor, Plate Factor, Colonies, Vol factor 50 ul plated, CFUs/ml, Average, StdDev; Trial 113 Water CEWAF 4-16: 4/26-29/16 Mote Exposure Trial 113. Adult Red drum water exposure to CEWAF. Microscreen Assay of Stock CEWAF 5/17/16. Performed six true replicates with two technical replicates of CEWAF with fish S9 liver enzyme activation for n = 12) – Sample (Positive control (MitC), Negative control (FSW), Reagents only, S9 mix & reagent, S9 mix & WAF, WAF only), Sample ID (null, CII-16-1924, CII-16-1925), Replicate, Replicate (1-1, 1-2, 2-1, 2-2, 3-1, 3-2), Plaques, volume factor, Fraction of plate, pfu/ml, average pfu/ml, Microscreen Assay CEWAF (% change over negative control), Microscreen Assay + Fish S9 enzyme activation (% change over negative control); Mote Exposure Trial 113 Microtox Acute Toxicity Assay Performed 7/21/16 (Two stock CEWAF vials were tested, did two true and two technical replicates from each for n = 8) – Replicate (CEWAF 1-1a, CEWAF 1-1b, CEWAF 1-2a, CEWAF 1-2b, CEWAF 2-1a, CEWAF 2-1b, CEWAF 2-2a, CEWAF 2.2b), Sample ID (CII-16-1924, CII-16-1925), Percent Inhibition of light production in comparison to control (%), Average, StDev; Mote Exposure Trial 113 – Bacterial (E. coli K-12/Microscreen) culture density after exposure to stock CEWAFs with and without S9 activation. Culture density measured by light absorbance at 600 nm wavelength (A600), measured for each sample (Mit C, Neg control (ASW), Reagents only, WAF S9 Activated, WAF + Reagents only, WAF only) and replicate (1-1, 1-2, 2-1, 2-2, 3-1, 3-2) combination; Trial 108 Water CEWAF 7-16: 6/28-30/16 Mote Exposure Trial 108. Adult Pompano water exposure to CEWAF (Acute exposure = spike with recovery). Microscreen Assay of Stock CEWAF 7/26/16. CEWAF "Spike 1" on 6/28 and "Spike 2" on 6/30/16. Controls are water from Non-exposure tanks, same day as CEWAF sample – Sample (Positive control (MitC), Negative control (FSW), Spike 1, carboy A&B, Spike 1, Carboy F, Control tank G (time of Spike 1), Spike 2, Carboy A&B, Spike 2, Carboy F, Control tank I (time of Spike 2)), Sample ID number (CII-16-2715, CII-16-2716, CII-16-2722, CII-27-2723, CII-16-2720, CII-16-2721, CII-16-2727, CII-16-2728, CII-16-2734, CII-16-2735, CII-16-2732, CII-16-2733), Replicate (1-1, 1-2, 2-1, 2-2, 1-1, 3-1, 3-2, 4-1), Dilution Factor, Plaques, volume factor, Fraction of plate, pfu/ml, average pfu/ml; 6/28-30/16 Mote Exposure Trial 108. Adult Pompano water exposure to CEWAF (Acute exposure = spike with recovery). Microtox Assay of Stock CEWAF and tank samples 7/26/16 Microtox Acute Toxicity Assay Performed 7/26/16 (Two stock CEWAF vials tested for each date, did two true and two technical replicates from each for n = 8) – CEWAF Stock Replicate (Stock 1 A&B 1a, Stock 1 A&B 1b, Stock 1 A&B 2a, Stock 1 A&B 2b, Stock 1 F 1a, Stock 1 F 1b, Stock 1 F 2a, Stock 1F 2b, Average), Sample ID (CII-16-2715, CII-16-2716, CII-16-2717, CII-16-2720, CII-16-2721, CII-16-2722, CII-16-2725, CII-16-2727, CII-16-2728, CII-16-2732, CII-16-2733, CII-16-2734, CII-16-2735, CII-27-2723), Percent Inhibition (%), Exposure Tank Replicate (Exposure tank A a, Exposure tank A b, Exposure tank E a, Exposure tank E b), Control Tank Replicate(Control Tank G 1a, Control Tank G 1b, Control Tank G 2a, Control Tank G 2b, Control Tank I 1a, Control Tank I 1b, Control Tank I 2a, Control Tank I 2b); Mote Exposure Trial 108 – Bacterial (E. coli K-12/Microscreen) culture density after exposure to stock CEWAFs and tank samples. Culture density measured by light absorbance at 600 nm wavelength (A600), measured for each sample (Mit C, Neg control (ASW), Spike 1. A&B, Spike 1. F, Spike 1. Control G, Spike 2. A&B, Spike 2. F, Spike 2. Control I) and replicate (1-1, 1-2, 2-1, 2-2) combination; MitC dose response: 11/1/16 Microscreen Assay- Mitomycin C Dose-Response – MitC dose (1 ug/ml, 0.5 ug/ml, 0.25 ug/ml, 0.125 ug/ml, 0.0625 ug/ml, 0 ug/ml), Replicate (1-1, 1-2, 2-1, 2-2, 1-1), dilution, plaques, ml conversion, fx plate, pfu/ml, Average, SD;

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