Emerging contaminants in marine invertebrates: concentrations and effects

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1 Emerging contaminants in marine invertebrates: concentrations and effects EMMA PRICHARD AND ELISE GRANEK ENVIRONMENTAL SCIENCE & MANAGEMENT, PORTLAND STATE UNIVERSITY, PORTLAND, OR, Collaborators: Kathy Conn 2, Bill Fish 1, Elena Nilsen 2, Angela Strecker 1, Lori Pillsbury 3, Steve Rumrill 4, 1 Environmental Science and Management, Portland State University 2 USGS Water Science Center (Tacoma and Portland) 3 OR Department of Environmental Quality 4 OR Department of Fish and Wildlife

concentrations in environment u Sub-lethal effects (DNA damage, enzymatic

2 Contaminants of emerging concern u Pharmaceuticals, personal care products (PPCPs), estrogenic compounds u Ubiquitous in aquatic systems u Low concentrations in environment u Sub-lethal effects (DNA damage, enzymatic alternations, organ-level effects i.e. foot detachment, gonadal alteration or damage.)

3 Sources of PPCPs to the Environment u Wastewater treatment plant discharge u Combined sewer overflows u Onsite wastewater treatment systems u Stormwater run-off u Illegal discharge u Agriculture and livestock u Vaporization (Breton and Boxall 2003, Daughton and Brooks 2011, Fong and Ford 2014)

4 Outline u Effects of PPCPs on marine and estuarine invertebrates (review) u Spatial and temporal variability of PPCP in Olympia oysters (Ostrea lurida)

5 Effects of PPCPs on marine invertebrates (review) Pharmaceuticals u have specifically designed modes of action u intended to illicit physiological response (Breton and Boxall 2003). Personal care products u Unintended adverse effects from interaction with non-target receptors

6 Current Literature u 16 papers on marine or estuarine invertebrates + PPCPs (~10% of total aquatic literature on PPCPs) u 42 total PPCPs u 25 active pharmaceutical ingredients (APIs) (60%), u 14 endocrine disrupting compounds (i.e., Bisphenol A) or estrogens (33%), u 3 personal care products (PCPs) (7%) u 12 different organisms u 10 Mollusca, 1 Arthropoda, 1 Rotifera u Mytilus galloprovincialis = most studied organism (56% of studies)

Biomarkers u Importance of Biomarkers u 61 different endpoints organismal organ- level i.e. Feeding rate, gamete release, LC50, mortality 9 different endpoints measured i.e. Organ- level bioaccumulanon, foot detachment 9 different endpoints measured cellular endpoints i.

7 Biomarkers u Importance of Biomarkers u 61 different endpoints organismal organ- level i.e. Feeding rate, gamete release, LC50, mortality 9 different endpoints measured i.e. Organ- level bioaccumulanon, foot detachment 9 different endpoints measured cellular endpoints i.e. Apoptosis, annbacterial acnvity 3 different endpoints measured subcellular (biochemical) i.e. AchE acnvity, camp, protein kinase C phosphorylanon 16 different endpoints measured subcellular (molecular) i.e. DNA damage, mrna expression, transcripnon levels 14 different endpoints measures subcellular (physiological) i.e. Lipofuscin intensity, phagocytosis, maldondialdehyde (MDA) content 10 different endpoints measured

Locations, Time, Levels u 17 study locations (6 locations from Adriatic Sea) u Temporal range from 1 hour to 30 days (mean of 90 hours) u Only 37% of studies ran

8 Locations, Time, Levels u 17 study locations (6 locations from Adriatic Sea) u Temporal range from 1 hour to 30 days (mean of 90 hours) u Only 37% of studies ran for a week or longer u 50% of PPCPs studied at environmentally relevant levels u ng/l- mg/l levels Geographic locations of studies cited in the literature review

9 LEVELS OF BIOLOGICAL Toxic chemicals can affect... COMPLEXITY Organ Functions Histological Lesions Cell Integrity & Metabolism Gene function Enzyme activity Membrane function Ecology & Growth Behavior & Reproduction Homeostasis Slide credit: Dr. Eugene Foster, Environmental Science and Management, Portland State University

Challenges in determining community and ecosystem effects u Gaps: u Extrapolating subcellular/organismal effects to community or ecosystem effects u Studies of effects on growth or reproduction

10 Challenges in determining community and ecosystem effects u Gaps: u Extrapolating subcellular/organismal effects to community or ecosystem effects u Studies of effects on growth or reproduction in any toxicological area Eggen, R.I., Behra, R., Burkhardt- Holm, P., Escher, B.I., Schweigert, N., Challenges in Ecotoxicology. Environ. Sci. Technol. 38, 58A 64A. doi: / es040349c

11 Key data gaps

Risk Management and Prioritization u Vast number of PPCPs and possible affected organisms u Prioritize probable ecologically relevant effects u Prioritize substances expected to pose greatest risk u

12 Risk Management and Prioritization u Vast number of PPCPs and possible affected organisms u Prioritize probable ecologically relevant effects u Prioritize substances expected to pose greatest risk u Use pharmaceutical mode of action to identify relevant biological effects Boxall, A.B.A., Rudd, M.A., Brooks, B.W., Caldwell, D.J., Choi, K., Hickmann, S., Innes, E., Ostapyk, K., Staveley, J.P., Verslycke, T., Ankley, G.T., Beazley, K.F., Review Pharmaceubcals and Personal Care Products in the Environment : What Are the Big Quesbons? 120,

13 Spatial and temporal variability of PPCPs in Olympia oysters (Ostrea lurida) Elise Granek, Kathy Conn, Elena Nilsen, Angela Strecker, Bill Fish, Lori Pillsbury, Steve Rumrill

14 Objectives u Identify spatial and temporal variability in O. lurida PPCP concentrations u Determine whether concentrations correlate with seawater and sediment levels u Identify contaminants at biologically relevant levels for O. lurida

Methods u Collect 60 Ostrea lurida per season (Summer, Winter, Spring) Coos and Netarts Bays u Oysters measured, shucked and weighed; guts rinsed u Aggregate

15 Methods u Collect 60 Ostrea lurida per season (Summer, Winter, Spring) Coos and Netarts Bays u Oysters measured, shucked and weighed; guts rinsed u Aggregate groups of 10 oysters analyzed for Lists 1 and 3 PPCPs (using LC-MS/MS) u Analyze temporal variability and correlations with seawater and sediment concentrations

16 List 1 List 1 - Acid Extraction in Positive Ionization Acetaminophen Digoxigenin Norfloxacin Sulfadimethoxine Ampicillin 1 Digoxin Norgestimate Sulfamerazine Azithromycin Diltiazem Ofloxacin Sulfamethazine Caffeine 1,7-Dimethylxanthine Ormetoprim Sulfamethizole Carbadox Diphenhydramine Oxacillin 1 Sulfamethoxazole Carbamazapine Enrofloxacin Oxolinic acid Sulfanilamide Cefotaxime Erythromycin-H20 Penicillin G 1 Sulfathiazole Ciprofloxacin Flumequine Penicillin V Thiabendazole Clarithromycin Fluoxetine Roxithromycin Trimethoprim Clinafloxacin Lincomycin Sarafloxacin Tylosin Cloxacillin 1 Lomefloxacin Sulfachloropyridazine Virginiamycin Dehydronifedipine Miconazole Sulfadiazine

17 List 3 List 3 - Acid Extraction in Negative Ionization Bisphenol A Furosemide Gemfibrozil Glipizide Glyburide Hydroclorothiazide 2-hydroxy-ibuprofen Ibuprofen Naproxen Triclocarban Triclosan Warfarin

18 Results : Summer 2013 and Spring 2014 Contaminant Coos Netarts Summer Spring Summer Spring Azithromycin 0/3 2/3 0/3 1/3 Diphenhydramine 3/3 0/3 0/3 0/3 Erythromycin- H2O 1/3 0/3 1/3 0/3 Furosemide 0/3 1/3 0/3 0/3 Hydrochlorothiazide 0/3 0/3 0/3 1/3 Ibuprofen 0/3 1/3 0/3 0/3 Naproxen 1/3 1/3 0/3 1/3 Sulfadiazine 2/3 0/3 1/3 0/3 Sulfamethoxazole 0/3 1/3 0/3 0/3 Virginiamycin M1 0/3 0/3 1/3 0/3

19 Results Contaminant Azithromycin, Erythromycin- H2O, Sulfamethoxazole Diphenhydramine Furosemide, Hydrochlorothiazide Ibuprofen, Naproxen Sulfadiazine Virginiamycin M1 Anbbiobc used for bacterial infecbons Anbhistamine (Benadryl) Diurebc edema, congenital heart failure, kidney disease Non- steroidal anb- inflammatory drug Sulfa drug- eliminates bacteria that cause infecbon, esp. UTIs Streptogramin anbbiobc; in fuel ethanol (prevent microbial contamin.); livestock rearing

20 Next steps u ODEQ results from 2013 sediment and water sampling of PPCPs u Spatial and temporal trends, correlations among fresh and marine waters, and marine bivalves u Mapping hotspots of and most common PPCPs

21 Future directions u Contaminants in septic outflow (sources to manage?) u Sampling to assess chronic vs. acute exposure (e.g., around storm events) u Examine additional sublethal effects (growth, reproductive output, larval survival) J. Botel

Acknowledgements u Funding from u Oregon Sea Grant Program Development Grant to EG; u Oregon Sea Grant Contaminants Grant to EFG, AS, BF, KC, EN, LP, SR; u OUS Katherine Bisbee II Grant to EG and AMF

22 Acknowledgements u Funding from u Oregon Sea Grant Program Development Grant to EG; u Oregon Sea Grant Contaminants Grant to EFG, AS, BF, KC, EN, LP, SR; u OUS Katherine Bisbee II Grant to EG and AMF u Jim Pankow Lab (Portland State University) for GC-MS analysis, and generous help from Lorne Isabelle and Wentai Luo u PSU students for help in the field and laboratory: Andy Harwood, Brianna Tarnower, Joey Peters, Dominic Galen, Dylan Dayrit u Gene Foster for use of his materials and with the review

EPA Method 1694: Analysis of Pharmaceuticals and Personal Care Products in Water

EPA Method 1694: Analysis of Pharmaceuticals and Personal Care Products in Water UCT Part Numbers ECHLD156-P ENVIRO-CLEAN HL DVB 500 mg, 6 ml cartridge VMF016GL 16 position glass block manifold VMFSTFR12