OCS Study • MMS 2006-008 Use of Biological Endpoints in Flatfish to Establish Sediment Quality Criteria for Polyaromatic Hydrocarbon Residues and Assess Remediation Strategies Final Technical Summary Final Study Report U.S. Department of the Interior Minerals Management Service Pacific OCS Region OCS Study MMS 2006-008 Use of Biological Endpoints in Flatfish to Establish Sediment Quality Criteria for Polyaromatic Hydrocarbon Residues and Assess Remediation Strategies Final Technical Summary Final Study Report Authors Luke A. Roy, Scott Steinert, Steve M. Bay, Darrin Greenstein, Yelena Sapozhnikova, Ola Bawardi, Cherlynn Seruto, Ira Leifer, Daniel Schlenk Principal Investigator Daniel Schlenk Prepared under MMS Cooperative Agreement No. 14-35-0001-31063 by Coastal Marine Institute Marine Science Institute University of California Santa Barbara, CA 93106-6150 U.S. Department of the Interior Minerals Management Service Camarillo Pacific OCS Region February 2006 Disclaimer This report has been reviewed by the Pacific Outer Continental Shelf Region, Minerals Management Service, U.S. Department of the Interior and approved for publication. The opinions, findings, conclusions, or recommendations in this report are those of the authors, and do not necessarily reflect the views and policies of the Minerals Management Service. Mention of trade names or commercial products does not constitute an endorsement or recommendation for use. This report has not been edited for conformity with Minerals Management Service editorial standards. Availability of Report Extra copies of the report may be obtained from: U.S. Dept. of the Interior Minerals Management Service Pacific OCS Region 770 Paseo Camarillo Camarillo, CA 93010 Phone: 805-389-7621 A PDF file of this report is available at: http://www.coastalresearchcenter.ucsb.edu/CMI/ Suggested Citation The suggested citation for this report is: Schlenk, D. Use of Biological Endpoints in Flatfish to Establish Sediment Quality Criteria for Polyaromatic Hydrocarbon Residues and Asses Remediation Strategies. MMS OCS Study 2006- 008. Coastal Research Center, Marine Science Institute, University of California, Santa Barbara, California. MMS Cooperative Agreement Number 14-35-0001-31063. 39 pages. Table of Contents FINAL TECHNICAL SUMMARY........................................................................ 1 FINAL STUDY REPORT....................................................................................... 5 1. Chapter 1 ........................................................................................................ 5 1.1 Introduction................................................................................................... 5 1.2. Materials and Methods.................................................................................. 6 1.2.1. Recirculating exposure system................................................ 6 1.2.2. Sediment chemical and water analysis .................................... 8 1.2.3. Biochemical endpoint measurements ...................................... 8 1.2.4. Statistical analysis.................................................................... 9 1.3. Results........................................................................................................... 9 1.3.1. Sediment PAHs........................................................................ 9 1.3.2. Biochemical markers of exposure............................................ 11 1.4. Discussion..................................................................................................... 14 1.5. Conclusion.................................................................................................... 17 2. Chapter 2 ........................................................................................................ 18 2.1. Introduction................................................................................................... 18 2.2 Materials and Methods.................................................................................. 19 2.2.1. Exposure system...................................................................... 19 2.2.2. Chemical evaluation of PAHs in sediments............................. 20 2.2.3. Biochemical endpoint measurements ...................................... 20 2.3 Results..................................................................................................... 21 2.4 Discussion..................................................................................................... 23 2.5 Conclusion .................................................................................................... 25 Acknowledgements ............................................................................................ 26 References........................................................................................................... 27 List of Tables Table 1. Concentrations of high molecular weight PAHs......................................... 9 Table 2. Concentrations of low molecular weight PAHs.......................................... 10 Table 3. E2 Concentrations (pmol/ml) in Hornyhead turbots................................... 14 List of Figures Figure 1. Recirculating system for exposing fish to sediments .............................. 7 Figure 2. High molecular weight PAHs measured in sediments ............................. 10 Figure 3. Low molecular weight PAHs measured in sediments.............................. 11 Figure 4. Equivalents of naphthalene, phenanthrene, and benzo(a)pyrene (µg/ml) in bile of hornyhead turbot (Pleuronichthys verticalis)....................................... 12 Figure 5. Percent DNA damage in comet tail .......................................................... 13 Figure 6. Tail Moment (TM) values for the blood and liver.................................... 13 Figure 7. Hepatic cytochrome P450 1A (CYP 1A) expression ............................... 21 Figure 8. FAC’s in halibut bile ................................................................................ 22 Figure 9. Plasma steroid concentrations between treatment groups ........................ 23 Use of Biological Endpoints in Flatfish to Establish Sediment Quality FINAL TECHNICAL SUMMARY STUDY TITLE: Use of Biological Endpoints in Flatfish to Establish Sediment Quality Criteria for Polyaromatic Hydrocarbon Residues and Assess Remediation Strategies. REPORT TITLE: Use of Biological Endpoints in Flatfish to Establish Sediment Quality Criteria for Polyaromatic Hydrocarbon Residues and Assess Remediation Strategies. CONTRACT NUMBER: 14-35-0001-31063 SPONSORING OCS REGION: Pacific APPLICABLE PLANNING AREA(S): Southern California FISCAL YEAR(S) OF PROJECT FUNDING: FY 01, FY 02 COMPLETION DATE OF REPORT: December 2005 COSTS: FY 01 - $64,746; FY 02 - $64,746; FY 03 - no cost CUMULATIVE PROJECT COST: $129,492 PROJECT MANAGER: Russell J. Schmitt AFFILIATION: Coastal Research Center, Marine Science Institute, University of California, Santa Barbara, CA 93106-6150 ADDRESS: University of California, Santa Barbara, CA 93106-6150 PRINCIPAL INVESTIGATOR: Daniel Schlenk ADDRESS: Department of Ecotoxicology and Environmental Toxicology, Science Laboratories I 316, University of California, Riverside, CA 92521 KEY WORDS: Polycyclic aromatic hydrocarbons, Pleuronichthys verticalis, Fluorescent aromatic compounds, DNA damage, PAHs, oil seep, CYP1A, FACs, GSI, sex steroids BACKGROUND: Input of Polyaromatic hydrocarbons (PAHs) occurs through anthropogenic and natural mechanisms. Toxicities resulting from chronic exposure include immune suppression, reproductive dysfunction and carcinogenesis. As most PAHs tend to be lipophilic, there is a high propensity for accumulation in organisms and sediments with high organic content. While analytical measurement of specific compounds has been shown to be a relevant indicator of exposure within invertebrates, rapid biotransformation prevents accurate assessments of exposure in vertebrates such as fish. 1 Final Technical Summary – Schlenk OBJECTIVES: The following study attempted to utilize biochemical and physiological indicators in flatfish to estimate a threshold concentration which could be used in risk assessment paradigms to evaluate sediments contaminated with PAHs. DESCRIPTION: Two species of flatfish were exposed to various dilutions of sediments collected from the natural oil seep off the coast of Santa Barbara, CA. In contrast to other studies carried out in anthropogenically contaminated areas, the predominant PAHs observed in the sediments were of low molecular weight. Hepatic cytochrome P450 1A (CYP1A), bilary fluorescent aromatic compounds (FACs), plasma steroid concentrations, gonadal somatic indices, and in some cases, hepatic DNA damage was utilized in each species exposed for either 7 (Hornyhead turbot) or 30 days (California Halibut) to diluted sediments. Attempts were made to generate dose-response curves which could be calibrated against reproductive function (GSI, sex steroids) for the estimation of sediment threshold concentrations. SIGNIFICANT CONCLUSIONS: Of all of the endpoints measured, diminished plasma concentrations of 17β-estradiol was the most sensitive endpoint in either species. With the exception of hepatic DNA damage in Hornyhead turbot following the 7 day exposure, none of the endpoints, in particular, the reproductive endpoints, displayed dose-response relationships preventing comparisons between indicators. Comparisons of a potential threshold at the 1% COP sediment dilution