SESSION 1: Innovative Approaches in Environmental Toxicology: Part I. Risk Assessment and Management.  (Chair, Sylvain DeGuise, sylvain.deguise@uconn.edu)

A Pilot Study to Evaluate the Potential Increase in River Water Toxicity Following Dam Removal.  Adria A. Elskus (aelskus@usgs.gov), U.S. Geological Survey, Maine Field Office, Orono, ME 04469; G. Mayer, C. Kim, L. LeBlanc, L.B. Perkins and R. Van Beneden, University of Maine, Orono, ME 04469.

Dramatic declines in Atlantic salmon (Salmo salar) populations have led to the complete loss of wild salmon in New England except for 8 rivers in Maine where salmon are now listed as endangered. Dam removal has been identified as the most important strategy for restoring salmon populations, but this can result in the release of contaminants from sediments into overlying waters. Because dams will be removed as part of the Penobscot River Restoration Project, there is a need to evaluate the toxic potential of Penobscot River sediments prior to dam removal. We are using a simple laboratory-based, sediment resuspension design and two well-established aquatic toxicology models, fathead minnows (Pimephales promelas) and zebrafish (Danio rerio), to evaluate if resuspension of Penobscot River sediment significantly elevates the toxicity of river water as measured by fish survival, hatch success, development, and immune competence, whether bioactive metals and/or endocrine disrupting substances are present, and to provide preliminary information on the types of chemicals likely to desorb during resuspension. River sediment resuspension did not increase organic pollutants (PAHs, PCBs, chlorinated pesticides) in our test system; metals data are pending. Preliminary results suggest river water alone elevates CYP1A expression compared to culture water; developmental abnormalities, vitellogenin, metallothionein expression and immune competence data are pending.

Contact: Dr. Adria A. Elskus U.S. Geological Survey, Maine Field Office, Dept of Biological Sciences 5751 Murray Hall, University of Maine Orono, ME 04469 Email: aelskus@usgs.gov

EVALUATION OF BUFFER ZONE EFFECTIVENESS IN THE MITIGATING THE RISKS OF AGRICULTURAL RUNOFF IN PRINCE EDWARD ISLAND.  Allison M. Dunn¹ (Allison.Dunn@EC.GC.CA), G. Julien¹, W. R. Ernst¹, A. Cook², K.G. Doe², and P.M. Jackman². ¹Environment Canada, Environmental Protection Branch, Dartmouth, NS; and ²Environment Canada, Environmental Conservation Branch, Moncton, NB.

Runoff from agricultural fields has lead to pesticide induced fish kills in Prince Edward Island (PEI). To minimize the risk posed by row crops, PEI introduced legislation in 2000 which stipulates a 10 metre buffer must be maintained between water courses and agricultural fields with slopes less than 5%. Since 2001, Environment Canada has been involved in a study to assess the effectiveness of 10 metre vegetative buffer zones in reducing toxicity, pesticide and nutrient loads to nearby aquatic ecosystems. Sample collectors have been placed in more than 40 fields at the field edge (0 m), 10 m and, on occasion, at distances greater than 10 m. Throughout the study’s duration, 17 rainfall events resulted in runoff collections; samples were collected within 24 hours and analysed for pesticides, water quality parameters and assayed for Daphnia magna toxicity. Buffer zones of 10 m were only moderately effective at reducing pesticide concentrations and toxicity and did not totally eliminate toxicity or decrease specific pesticide or unionized ammonia concentrations below the published LC50 value for Daphnia magna. Elevated pesticide concentrations and toxicity may be due to such factors as slope length and/or row orientation, land management practices or contribution from sprayer track rows, posited to concentrate and channel flow.

Contact: Allison M. Dunn, Environment Canada, Environmental Protection Branch, Dartmouth, NS.  Email: Allison.Dunn@EC.GC.CA

INTERPRETATION OF UNCERTAINTY IN DIOXIN/FURAN DATA FOR THE RISK ASSESSOR AND ENVIRONMENTAL DECISION-MAKER.  Nancy C. Rothman, New Environmental Horizons, Inc., 34 Pheasant Run Dr., Skillman, NJ 08558 and Susan D. Chapnick (s.chapnick@comcast.net), New Environmental Horizons, Inc., 2 Farmers Cir., Arlington, MA 02474.

What is the difference between low resolution and high resolution dioxin/furan results? What are “EDLs” and how do they differ from a sample reporting limit? Do EDLs have uncertainty? If so, what is the source of the uncertainty and are the non-detected results usable for risk assessment? Do detected dioxin/furan results have uncertainty? Will this uncertainty affect the way the data can be used in quantitative risk assessment? What is the “TEQ” and how is it derived? These are key questions that an environmental decision-maker might ask in reviewing dioxin/furan results to compare to regulatory standards or to evaluate potential risk to human health or the environment. We will answer these questions and give key information concerning dioxin/furan analysis and interpretation of results in terms of potential uncertainty that may bias the data. Topics to be covered will include method issues that may affect bias in results, reporting limit uncertainties (where they come from and why this is a critical issue for risk assessors), and homologues versus specific isomers in terms of toxicity interpretations. Examples from case studies and recommendations to reduce dioxin/furan data uncertainty will be given throughout the discussion.

Contact: Susan D. Chapnick, New Environmental Horizons, Inc., 2 Farmers Circle, Arlington, MA 02474. Email: s.chapnick@comcast.net

SESSION 2: Innovative Approaches in Environmental Toxicology: Part II. Toxicity Testing & Ecotoxicology.  (Chair, Sylvain DeGuise, sylvain.deguise@uconn.edu)

*MERCURY LEVELS IN WILD-GROWING FUNGI FROM INTERIOR AND COASTAL FORESTS NEAR THE BAY OF FUNDY, CANADA.  Mina Nasr (c4a33@unb.ca) and Paul A. Arp, Faculty of Forestry and Environmental Management, UNB, Fredericton, N.B., Canada, E3B 6C2.

Contributions of wet and dry atmospheric deposition of mercury (Hg) may have led to elevated amounts of Hg inputs into the coastal forests of the Bay of Fundy. Within these forests, soil-based fungi likely accumulate the deposited Hg within their mycelium and fruiting bodies (mushrooms). In this study, Hg concentration in mushrooms varied widely (15 to 5,500 ppb dry matter), mostly depending on fungal species, site location (interior versus coastal forests) and conditions (forest litter/ soil type). On average, wild-growing mushrooms from upland forests in Grand Manan Island showed higher Hg concentrations than those from Point Lepreau (coastland), and from Fredericton in central New Brunswick within the same species. Some of this Hg was in the form of neurotoxic methyl Hg (2 to 50 %). Fruiting bodies of Boletus edulis, Cortinarius armillatus and Bankera canosa contained the highest Hg concentrations, i.e. 3700, 2600 and 2290 ppb dry matter, respectively. The concentration ratio of Hg in cap to stalk was 2.5 in Amanita muscaria and Russula peckii. The ratio of Hg in mushroom caps to Hg in the soil substrate was approximately 8. Overall, certain ubiquitous mushroom species could become useful indicators of Hg bio-availability in the soil, regardless of Hg source. As such, mushrooms are also a component of the terrestrial bio-cycling and local food chains.

Contact: Mina Nasr Faculty of Forestry and Environmental Management, 28 Dineen Drive, University of New Brunswick, Fredericton, NB, Canada E3B 6C2. Email: minab.nasr@yahoo.com

*ANALYSIS OF ZEBRAFISH EXPOSED TO SYNTHETIC HORMONES.   Emily Notch (emily_notch@umit.maine.edu), Danielle Miniutti, Greg Mayer, Biochemistry, Microbiology & Molecular Biology Department, University of Maine Orono.

Recently, much attention has been given to endocrine active compounds in the environment. One class of xenoestrogens includes the synthetic estrogens commonly found in pharmaceuticals such as oral contraceptives and hormone replacement therapies. Synthetic estrogens such as these mimic natural estrogens well at the receptor level, but are more resistant to degradation by natural processes. Because of its greater stability and higher potency in vivo, ethinylestradiol (EE2) may be of disproportional toxicological importance despite being found at much lower concentrations than natural steroids such as E2 and E1. We exposed adult zebrafish to waterborne concentrations of EE2 and utilized fluorescence based quantitative RT-PCR to examine predictable biomarkers of endocrine disruption. We also examined the potential for EE2 to produce other deleterious effects in aquatic organisms, including disruption of cellular processes such as DNA repair, specifically nucleotide excision repair that removes a variety of environmentally relevant adducts.

Contact: Emily Notch University of Maine, 5735 Hitchner Hall Orono, ME 04469 Email: emily.notch@umit.maine.edu

*Avoidance response of the marine snail, Ilyanassa obsoleta, to harbour sediments.   Steve Marklevitz¹ (smarklev@dal.ca), J. Hellou^{2, 3}, ¹Marine Biology and Oceanography Departments, Dalhousie University, Halifax, Nova Scotia, Canada. ²Bedford Institute of Oceanography, Department of Fisheries and Oceans, Dartmouth, Nova Scotia, Canada; ³Chemistry and Oceanography Departments, Dalhousie University, Halifax, Nova Scotia.

Harbours receive multiple sources of contamination where sediments act as a sink for natural products and anthropogenic chemicals. Polycyclic aromatic hydrocarbons (PAH) were previously analysed in Halifax Harbour and levels of many were above the Canadian Council of the Marine Environment sediment quality guidelines. A widespread benthic species could not be easily collected from field sediments. Therefore, laboratory exposures were developed to assess the quality of harbour sediments. The horizontal movement of marine snails described as an avoidance/preference response relative to contaminated/reference sediments was investigated. The behavioural response of animals is a sensitive, cumulative biological effect, observed at the organism level. Behavioural changes can result from exposure to neuro-toxic chemicals or contaminants with other modes of action. The sensitivity of the behavioural response can vary with species and type of chemical, but is expected to be more sensitive than an acute narcotic type response. An avoidance of sediments can translate into population extinction or be viewed as a defense mechanism. Experiments with mud whelks Ilyanassa obsoleta yielded distinct and changing avoidance behaviour, with time, towards contaminated sediments. The response was compared to PAH levels in the sediments. Significant avoidance was observed after 24 to 72 hrs of exposure. For lower percentage of harbour in reference sediments, there was a preference for contaminated sediments, while for higher levels of harbour in reference sediments, there was an avoidance of contaminated sediments. The reduction in avoidance is proposed to be in response to the balance between food and contaminants availability in the sediments.

Contact: Stephen Marklevitz, 1472 Tower Rd, Halifax, Nova Scotia, Canada, B3H 4K8 Email: smarklev@dal.ca

*APPLICATIONS AND ASSUMPTIONS OF STABLE ISOTOPE ANALYSIS IN ECOTOXICOLOGY.  Timothy D. Jardine (m614u@unb.ca), Karen A. Kidd, Canadian Rivers Institute and Biology Department, University of New Brunswick, Saint John, NB, Canada, and Aaron T. Fisk, Warnell School of Forestry and Natural Resources, University of Georgia, Athens, GA.

Stable isotope analysis (SIA) has become a powerful tool for ecotoxicologists to study dietary exposure and bioaccumulation of contaminants in wild animal populations. The use of SIA in ecotoxicology continues to expand, and while much more is known about the mechanisms driving patterns of isotope ratios in consumers, there remain several assumptions that can influence interpretation of data from field studies. Current uses of SIA in ecotoxicology will be outlined, including estimating the importance of dietary sources of carbon and their application in bioaccumulation studies, and six key assumptions associated with the approach and related caveats will be presented including: 1) diet-tissue fractionation is equal among species, 2) within a given species, diet-tissue fractionation never changes, 3) different tissues accurately reflect diet, 4) isotope ratios in a single species are comparable across systems, 5) omnivory is uncommon, and 6) all species are part of the local food web. We advocate that when studying multiple systems, the most important component in study design with SIA is an adequate characterization of baseline isotopic signatures, which can account for a d15N range of up to 12‰ across systems. We also advocate that the life-history characteristics (e.g. migratory habits, diet variation, and reproductive cycles) of the study species may play an important role in the interpretation of contaminant and isotope data. A thorough appreciation of the assumptions above will strengthen our ability to effectively use SIA to gain new insights in ecotoxicology.

Contact: Timothy D. Jardine Canadian Rivers Institute and Department of Biology, Loring Bailey Hall, University of New Brunswick, Fredericton, NB, Canada E3B 5A3. Email: tim.jardine@unb.ca

*MODELING ECOLOGICAL PATHWAYS OF MERCURY THROUGH AQUACULTURE.  Laura B. Sweeney (laura.sweeney@unb.ca), M.D.B. Burt, A. Diamond, P.A. Arp, University of New Brunswick, Fredericton, Canada.

Attention has been focused on food safety and the nutritional value of farmed finfish products because of increasing global demands on aquaculture. Our research has the goal of determining inputs of Hg, a heavy metal of concern, into the aquaculture cycle and modeling resulting Hg through-puts. In this context, we present a bio-energetics model to quantify biomass and Hg accumulation in farmed fish, from feed to fish, to waste. In collaboration with several fish farms in New Brunswick, fish, feed, and waste samples were collected on a regular basis and analyzed for total Hg. In addition, laboratory trials were conducted to determine the rate of administered methyl Hg absorption and release from farmed Atlantic haddock under controlled tank conditions. This was done for model parameterization and calibration. We now have determined that Hg concentrations in locally derived fish feed are not significantly different from Hg concentrations in internationally produced diets, with values ranging from 14ppb to 56ppb (dry wt, p<0.01). Compared with wild Atlantic salmon, Hg concentrations in farmed fish remained consistently low with increasing fish size, but wild fish concentrations increased (respective means, 84ppb and 260 ppb (dry wt, p<0.01). We have also determined that Hg, once administered to the fish in form of a fixed dose, is distributed throughout fish from the liver, to a gradual release into muscle tissue, where Hg will reside with a half-life of about 70 days.

Contact: Laura Sweeney 340 Victoria St., Apt 2 Fredericton, NB E3B 1W6 e-mail: laura.sweeney@unb.ca

SESSION 3: Natural Resource Damage Assessment (NRDA) (Chair, Stephen E. Petron, Stephen.Petron@ch2m.com)

INNOVATIVE APPROACHES TO PROVIDING RESTORATION FOR SMALL SPILLS.   Kate Clark (Kate.Clark@noaa.gov) NOAA Office of Response and Restoration (ORR), 28 Tarzwell Drive, Narragansett, RI 02882, T. Brosnan, NOAA ORR, 1305 East-West Highway, Silver Spring, MD, 20910.

To cost-effectively provide restoration for small spills, natural resource Trustees are continually developing alternative methods for assessing and restoring injured resources. Some practitioners consider a spill of less than 50,000 gallons to be a ‘small spill.’ However, a ‘small spill’ cannot be defined solely by the volume of the release since the ecological sensitivity of an area must also be considered. ‘Small spills,’ as discussed in this presentation, are spills in which the cost and time required to implement a Natural Resource Damage Assessment (NRDA) under the Oil Pollution Act of 1990 (OPA) would ultimately outweigh the cost and size of the compensatory restoration required to restore the injured natural resource. To cost-effectively provide restoration for small spills, some states have developed compensation tables, cost-per-acre habitat analysis, penalty schedules, and consent agreements as way of expediting small spill settlements. Conservative assumptions and scaling models have been used in lieu of lengthy and costly injury assessment field studies. Further, when in-kind or in-place restoration of urbanized waterways is not always feasible, alternative approaches to restoration can be sought. This presentation discusses three areas in which small spill NRDAs have been expedited: 1) invocation of state regulatory authorities to address restoration, 2) the use of cooperative expedited injury assessment methods (i.e., reasonably conservative injury estimates, habitat equivalency analysis, Type A modeling), and 3) alternative restoration approaches.

Contact:  Kate Clark, Injury Assessment Coordinator, NOAA Office of Response and Restoration Narragansett Lab, 28 Tarzwell Dr., Narragansett, RI 02882.  Email: kate.clark@noaa.gov

NATURAL RESOURCE DAMAGE ASSESSMENTS:  HOW COOPERATIVE ASSESSMENTS YIELD SIGNIFICANT RETURNS.  Steve Davis (SDavis@entrix.com), A. Parsons, and T. Williams, ENTRIX, Inc., Augusta, ME., Methuen, MA., and Barrington, Il.

A cooperative Natural Resource Damage Assessment (NRDA) is one where Natural Resource Trustees (Trustees) and Responsible Parties (RPs) are jointly involved in conducting a natural resource injury assessment and planning/implementing restoration. The degree of cooperation can vary from Trustees simply sharing their assessment plans and data to a fully integrated process where the Trustee/RP team jointly designs and conducts investigative studies and restoration measures.

In 2005, Maine Trustees, a paper manufacturing company, and local non-governmental organization (NGO) jointly participated in developing a creative natural resource damage settlement for a site located in Westbrook, Maine. A cooperative NRDA was undertaken to evaluate natural resource injury at the site, create restoration alternatives that fit various regulatory expectations, and allow for long-term management of the selected restoration alternative by an NGO. This settlement involved developing compensation parameters for groundwater injuries. A cooperative assessment and resulting partnership resulted in resolution of the RPs NRDA liability, compensation for natural resource injuries to Maine trust resources, and enhancement and dedication of habitat along the Presumpscot River.

This cooperative NRDA resulted in cost savings, reduced potential for litigation and shortened time to restoration. This presentation will provide site background information, a depiction of the cooperative NRDA framework and process, various parties involved, along with the details of the resulting settlement and selected restoration project.

Contact: Steve Davis, ENTRIX, Inc., Augusta, Maine.  Email: SDavis@entrix.com

NOAA’S APPROACH TO COOPERATIVELY RESOLVING NATURAL RESOURCE LIABILITY.   Kenneth Finkelstein (Ken.Finkelstein@noaa.gov), NOAA Office of Response and Restoration, c/o EPA Region 1 (HIO), 1 Congress Street, Boston, MA 02114.

NOAA’s Office of Response and Restoration (OR&R) acts as a federal natural resource trustee whose principal mission is to protect and restore natural resources by countering and responding to environmental threats and promoting sound decision-making in the coastal zone. The integration of remedial and restoration investigations, planning, and activities is a successful paradigm for achieving restoration-based settlements. Because NOAA places a technical liaison respectively within or nearby federal and state remedial decision-makers, we effectively work to integrate the remedial and damage assessment process to protect natural resources, to reduce or eliminate residual natural resource injuries after cleanup, and to achieve restoration as part of a cooperative natural resource injury settlement with responsible parties.

There are two primary paths within OR&R that can lead to the resolution of natural resource damage liability under CERCLA; 1) a cooperative and integrated approach to remediation and restoration leading to a negotiated comprehensive settlement, the results of which are often included in the Consent Decree or 2) a formal damage assessment to assess injury that uses the codified regulations under 43 CFR Part 11. Both processes result in a Restoration Plan but the former provides the benefits of flexibility, speed, and cost efficiency. Nevertheless, the latter may be necessary when: 1) the responsible party is not cooperative, 2) there is a potential for large-scale economic and biological injury, or 3) the remedial action does not address significant natural resource injury. A similar approach is available at oil spills whereby the responsible party is invited to engage in natural resource injury assessment and restoration efforts under Trustee oversight.

The cooperative integrated remediation/restoration approach is characterized by 1) an integrated and streamlined process for data assembly to support planning for remediation and restoration, 2) development of protective cleanup strategies to minimize residual injury and enhance recovery of coastal areas, 3) injury assessment, restoration planning and scaling, 4) the negotiated release of natural resource liability through a Covenant Not To Sue, and 5) implementation and monitoring of restoration.

Cooperative comprehensive or universal settlements to resolve both cleanup and natural resource liability under CERCLA are becoming more common around the country. This presentation will include a discussion of such settlements in Connecticut, Rhode Island, and Massachusetts.

Contact:  Kenneth Finkelstein, NOAA Office of Response and Restoration, c/o EPA Region 1 (HIO) 1 Congress Street, Boston, MA 02114.  Email: Ken.Finkelstein@NOAA.gov

ESTIMATION OF NATURAL RESOURCE DAMAGES FOR 23 FLORIDA CASES USING MODELING OF PHYSICAL FATES AND BIOLOGICAL INJURIES.  Jill Rowe (jrowe@appsci.com), Deborah French McCay, and Nicole Whittier; Applied Science Associates, Inc., 70 Dean Knauss Drive, Narragansett, RI 02882-1143.

Oil spill modeling was performed using the “type A” (simplified, with a minimum of field data requirements) modeling approach and SIMAP (Spill Impact Model Analysis Package) to assess marine biological injuries and natural resource damages (NRD) for 23 spill cases identified by Florida Department of Environmental Protection (DEP). The objectives were to provide (1) an assessment of the pathways and fate of the oil, and thus, estimate exposure to the water surface, shoreline and other habitats, water column, and sediments; and (2) estimates of injuries to wildlife, aquatic organisms, and habitats that were used to scale compensatory restoration. Compensatory habitat restoration for all quantifiable wildlife, fish and invertebrate injuries was scaled using Habitat Equivalency Analysis (HEA) and wetland habitat creation as mitigation. However, this is not a direct method of increasing sea turtle production. Therefore, scaling was performed to estimate the number of hatchlings needed to compensate for the sea turtle injuries. DEP then successfully submitted claims for NRD to the US Coast Guard Oil Pollution Fund.

Contact: Jill Rowe Applied Science Associates, Inc. 70 Dean Knauss Drive Narragansett, RI 02882-1143.  Email:  jrowe@appsci.com

NATURAL RESOURCE DAMAGE ASSESSMENTS IN URBANIZED AND CHANGING HABITATS - BERRY'S CREEK, NJ.  Mark S Laska (mlaska@geeinc.net), Great Eastern Ecology, Inc., 2231 Broadway, New York, NY 10024, Richard W. Galloway, Honeywell International, Morristown NJ, and Daniel T. Guest, MACTEC, Hamilton NJ.

Natural Resource Damage (NRD) penalties follow from CERCLA or OPA actions where a release into the environment has resulted in a loss of ecological services. Habitat Equivalency Analysis (HEA) is a model that is used to estimate the extent to which the PRPs are required to reimburse the public for the injury, payable in the form of cash or equivalent habitat enhancements on site or elsewhere. The HEA analysis is based on the presumption that before the release a baseline level of habitat services existed. However, complex urban industrial areas have experienced generations of habitat service losses resulting from development as well as releases. CERCLA is specific that a PRP inherits the environmental degradation of previous property owners, but where is that baseline to be drawn with regards to habitat? For instance, the habitat of the New Jersey Meadowlands has changed dramatically in the last 150 years. The remaining habitat in Berry's Creek, the location for a recent CERCLA action, was formerly freshwater Atlantic white cedar forest, and then it became brackish Spartina alterniflora marsh, and is now primarily Phragmites-dominated marsh. Wetlands have been drained and filled for navigational and developmental uses diminishing habitat value. We examine the difficulties and complexities in establishing a HEA protocol when baseline habitat values are changing over time and are due to multiple influences, releases and non-release events.

Contact: Mark S. Laska, Ph.D., Great Eastern Ecology, Inc. 2231 Broadway, New York, NY 10024 Email: mlaska@geeinc.net

SESSION 4: What’s New in the Gulf of Maine? (Chair, Janet Robinson, jrobinson@woodardcurran.com)

BACTERIAL POLLUTION SOURCE TRACKING IN COASTAL NEW HAMPSHIRE AND MAINE.  Steve H. Jones (shj@cisunix.unh.edu), University of New Hampshire, Durham, NH, 03824; N. Landry and S. Sumner, NH Department of Environmental Services, Concord, NH 03301.

Ocean beaches and shellfish harvesting are enormously popular recreational activities in New Hampshire. With increasing population growth and coastal development, additional pollution sources and documented impacts to beach water quality constitute an increasingly significant threat to public health. Investigations for identifying pollution sources are often inconclusive, and when sources are identified and eliminated, water quality has not always been improved.

The use of Escherichia coli ribotyping has provided critical information on the most significant source species in numerous studies conducted in the NH Seacoast and the southern Maine coast. Ribotyping has been conducted using a RiboPrinter, and fecal and water samples from beaches, estuaries and freshwater tributaries have been collected in cooperation with state agencies and other volunteers. A large regional (ME, NH, VT, MA) library and study site-specific small libraries of known source ribopatterns from 35 species have been used to identify source species for water isolates. Studies have included research to test method assumptions and surveys to identify pollution sources in areas of concern. Results show humans, pets, wild animal and livestock species to be significant types of pollution sources in different coastal areas. Use of E. coli ribotyping has become progressively more focused and integrated into more comprehensive MST efforts to open more shellfish harvesting areas and beaches by identifying and eliminating pollution sources.

Contact: Stephen Jones Jackson Estuarine Laboratory, 85 Adams Point Rd., Durham, NH 03824 e-mail: shj@unh.edu

MERCURY LEVELS IN SEABIRDS IN THE GULF OF MAINE.  W. Goodale¹ (wing.goodale@briloon.org), D. Evers¹, B. Allen², J. Ellis³, S. Hall⁴, S. Kress⁴,  S. Mierzykowski⁵, and L. Welch⁶; ¹BioDiversity Research Institute, ²Maine Inland Fisheries and Wildlife, ³Shoals Marine Laboratory, ⁴National Audubon Society, ⁵U.S. Fish and Wildlife Service, ⁶Maine Coastal Islands NWR.

We conducted a pilot study to screen mercury levels in Gulf of Maine seabirds in an effort to determine which species are most at risk, are the most appropriate bioindicators, and to refine sampling methods. From 13 Gulf of Maine islands, we evaluated Hg levels in the eggs or blood of seven species of seabirds: razorbill (Alca torda), black guillemot (Cepphus grille), Atlantic puffin (Fratercula arctica), double-crested cormorant (Phalacrocorax auritus), common eider (Somateria mollissima), Leach’s storm-petrel (Oceanodroma leucorhoa), and common tern (Sterna hirundo). We found: (1) black-guillemots and double-crested cormorants had the highest Hg levels; (2) a suggestive relationship between cormorant levels and proximity to river outflows, but not statistically significant; (3) no significant relationship between cormorant Hg levels and proximity to the mainland; (4) low cormorant intra-clutch and island variation; and (5) a significant relationship between within cormorant clutch mean egg Hg levels and standard deviation. These results indicate that black guillemots, double-crested cormorants, and Leach’s storm-petrels are effective bioindicators of Hg and other containments in the marine environment.

Contact: Wing Goodale, BioDiversity Research Institute, 19 Flaggy Meadow Road, Gorham, ME 04038 URL: www.briloon.org Email: wing.goodale@briloon.org

CONCENTRATION OF TOXIC POLLUTANTS IN CASCO BAY SEDIMENTS AND BLUE MUSSELS AS INDICATORS OF ECOSYSTEM CONTAMINATION.  Diane Gould (gould.diane@epa.gov), US Environmental Protection Agency and Lee Doggett, Maine Department of Environmental Protection.

Casco Bay Estuary Project sampled the Bay’s bottom sediments in the early 1990s and again in 2000/2001 for heavy metals, pesticides, PAHs, polychlorinated biphenyls (PCBs), butyltins, dioxins (organochlorines) and furans. The results of the earlier sampling indicated that toxics were found throughout the Bay and that “high” concentrations (based on the NOAA Status and Trends Program standards) occurred in primarily in the Inner Bay, the area nearest to Portland, with some hot spots in other parts of the Bay. Analysis of the 2000/2001 data indicates that total pesticides, tributyl tin, total PCBs and most trace metals decreased in the Bay over the past decade. PAHs showed some localized increases. In general, the concentrations of toxics in the Bay appear to be decreasing or staying the same. To assess the impact of toxic pollution on biota, CBEP summarized blue mussel monitoring data from Maine DEP and the CBEP. Mussels from most areas in the Bay do not have concentrations elevated above those of “pollution-free” reference sites, with the exception of selected heavy metals in some developed sites (e.g., Mill Creek, Great Diamond Island and the inner Fore River). Also, PAHs are highly elevated in the Inner Fore River.

Contact: Diane Myrick Gould, Ph.D. USEPA 1 Congress Street, Suite 1100 COP Boston, MA 02114.  Email:  gould.diane@epa.gov

SESSION 5: Spatial Ecology in Ecological Risk Assessment (Chair, Anne Kuhn-Hines, kuhn.anne@epa.gov)

Forensic Mapping: Using GEOGRAPHIC INFORMATION SYSTEMS (gis) to track environmental contaminants.  Kirsten Read Boettcher (kirsten@orbismaps.com) & Rosemary Mosher, Orbis, LLC, 19 Hyde Road, PO Box 1105, Gray, Maine 04039.

Recent advances in GIS have added new dimensions to environmental research and analysis. Forensic Mapping is an innovative GIS application which layers both historical and contemporary land-use with environmental datasets and map sources to create a dynamic, queriable database. These databases translate into quantifiable, interactive, digital maps that serve as an integrated information clearinghouse that can be archived, managed and presented graphically. The digital maps allow for visual tracking of historical land use patterns, identification of potential contaminant source areas and human and ecological receptors through space and time. The historical development of the Greater Portland Peninsula (ME) provides a typical case study for the integration of land use investigations and historical forensics. Like many urban landscapes, Portland experienced extensive industrialization in the 19th and 20th centuries. The industrial chemicals used in many of these business activities have left a lasting mark on the environment. This case study examines and maps specific industries and related chemicals as an example of how complex visualization of data through time and space can aid in creating a more complete understanding of the environmental setting.

Contact:  Kirsten Read Boettcher, 19 Hyde Road, PO Box 1105, Gray, Maine 04039. Email: kirsten@orbismaps.com

EVALUATING EFFECTS OF LOW QUALITY HABITATS ON REGIONAL POPULATION GROWTH IN PEROMYSCUS LEUCOPUS: INSIGHTS FROM FIELD-PARAMETERIZED SPATIAL MATRIX MODELS.  Jason S. Grear¹ (grear.jason@epa.gov) and Catherine E. Burns² (catherine.burns@yale.edu), ¹U.S. Environmental Protection Agency, Atlantic Ecology Division, 27 Tarzwell Drive, Narragansett, RI 02882 and ²Department of Ecology & Evolutionary Biology Yale University, 165 Prospect St., New Haven, CT 06511.

Due to complex population dynamics and source-sink metapopulation processes, animal fitness sometimes varies across landscapes in ways that cannot be deduced from simple density patterns. In this study, we examine spatial patterns in fitness using a combination of intensive field-based analyses of demography and migration and spatial matrix models of white-footed mouse (Peromyscus leucopus) population dynamics. We interpret asymptotic population growth rates from these spatial models as fitness-based measures of habitat-quality and use elasticity analysis to further explore model behavior and the roles of migration. In addition, we compare population growth rates at the spatial scale of single habitats and the landscape-level scale at which these habitats are assembled. To this end, we employ emerging techniques in multi-scale estimation of demography and movement and recently described vec-permutation methods for spatial matrix notation and analysis. Our findings indicate that the loss of low quality habitats or reductions in movement from these habitat into higher quality areas could negatively affect landscape-level population fitness.

Contact: Jason S. Grear, U.S. Environmental Protection Agency, Atlantic Ecology Division, 27 Tarzwell Drive, Narragansett, RI 02882.  Email: grear.jason@epa.gov

UTILIZING BROAD SCALE ENVIRONMENTAL MONITORING DATA TO PREDICT ESTUARINE SEDIMENT METAL CONCENTRATIONS ALONG THE UNITED STATES’ ATLANTIC COAST.  Jeffrey W. Hollister (hollister.jeffrey@epa.gov), Department of Natural Resources Science, University of Rhode Island, Kingston, RI  02881,  Peter V. August, Department of Natural Resources Science, University of Rhode Island, Kingston, RI  02881, and John F. Paul, Environmental Protection Agency, Office of Research and Development, Research Triangle Park, NC 27711.

Both point and non-point sources of pollution degrade the ecological condition of estuarine receiving waters by adding sediments, nutrients, pesticides, and toxic metals. Many studies have documented strong quantitative relationships between the anthropogenic sources of these pollutants and resultant declines in ecological condition indicating it is possible to build reliable, predictive models of estuarine condition that may be used to identify potentially impacted estuaries. As such, we developed a total of 45 predictive models for the total concentrations of copper, lead, mercury, and zinc in estuarine sediments along the Southern New England and Mid-Atlantic regions of the United States. We constructed the models using landscape structure, estuarine properties, and sediment metal inputs as predictors. Using Information Theoretic assessment and model-averaging approaches, we assessed these variables and models and found total developed land and percent silt/clay of estuarine sediment were the most important variables for predicting the presence of all 4 metals. Estuary area, river flow, tidal range, and total agricultural land varied in their importance. The model-averaged predictions explained 78.4%, 70.5%, 56.4%, and 50.3% of the variation for copper, lead, mercury, and cadmium respectively. Overall prediction accuracies of the Effect's Range values were 83.9%, 84.8%, 78.6%, 92.0% for copper, lead, mercury, and cadmium respectively. From these results, we conclude that broad scale predictive models built from existing monitoring data provide reliable estimates of impacted estuaries and these impacts are best described by the physical characteristics of the estuarine sediment and the total amount of developed land in adjacent watersheds.

Contact:  Jeffrey W. Hollister, Department of Natural Resources Science University of Rhode Island, Kingston, RI 02881. Current and Corresponding Address: Science Department, U.S. Coast Guard Academy, New London, CT 06320. After 01 June 2006, Email contact at hollister.jeffery@epa.gov

CONSIDERING HABITAT, BEHAVIORS AND SPATIAL INTERACTIONS WITHIN TERRESTRIAL ECOLOGICAL EXPOSURE ASSESSMENT:  A CASE STUDY APPLICATION OF THE SPATIALLY EXPLICIT EXPOSURE MODEL (SEEM).  Wickwire W,T.¹ (wickwire@menziecura.com), Johnson MS², Parsons PJ³, Burmistrov D¹, and Menzie CA¹.  ¹Menzie-Cura & Associates, Inc., Winchester, MA. ²U.S. Army Center for Health Promotion and Preventive Medicine, Aberdeen Proving Ground, MD. ³Wadsworth Center, New York State Department of Health, Albany, NY.

Quantifying exposure of terrestrial wildlife to soil-based contaminants historically relies on simplified assumptions about foraging areas and habitat preferences. Over the past few years, spatial modelers have developed exposure models accounting for the impact of wildlife behaviors on overall exposure; however, few have been field tested. The Spatially Explicit Exposure Model (SEEM) has been developed to combine wildlife habitat preferences, foraging behaviors and chemical distributions to estimate population-level wildlife exposures. Within SEEM, individuals move across a predefined landscape (chemistry and habitat suitability) guided by habitat preferences and foraging ranges. Food consumption events occur for a user-defined number of times per day. Spatial models may not always yield predictable results. In order to evaluate how SEEM operates with measured data, the impact of lead in soil on songbirds at two small arms ranges is explored. Small arms ranges are a necessary and important component of US Army bases. SEEM is a powerful tool for examining the impact of soil lead distributions on songbirds moving through varying habitats surrounding the ranges. Measured data supply the necessary model inputs and provide a useful comparison to model outputs. Spatial models are accessible and increase the power of exposure assessments.

Contact: W.T. Wickwire, Menzie-Cura & Associates, Inc., Winchester, MA.  Email: wickwire@menziecura.com
 

SESSION 6: Application of Molecular Approaches and Tools to Ecological Risk Assessment (Chair, Diane Nacci, nacci.diane@epa.gov)

Application of Genomic Technologies to Ecological Risk Assessment.  Joshua W. Hamilon, Professor Pharmacology and Toxicology, Dartmouth College, New Hampshire.

Molecular evolution and molecular modeling of ecotoxicologically relevant CYP1A genes.  Jared Goldstone¹ (jgoldstone@whoi.edu), Jahnavi Prasad², Sandor Vajda², and John Stegeman¹; ¹Biology, Woods Hole Oceanographic Institution and ²Department of Biomedical Engineering, Boston University.

Cytochrome P450 family 1 (CYP1) genes are prominent in response to and metabolism of environmental contaminants. Toxic mechanisms of CYP1A include both metabolic activation of polynuclear aromatic hydrocarbons (PAHs) to DNA-adducting compounds and uncoupling by polyhalogenated aromatic hydrocarbons (PHAH) to produce reactive oxygen. We are examining the origin of CYP1 genes by genome searching, gene cloning, and phylogenetic analyses in aquatic organisms, including the sea urchin Strongylocentrotus purpuratus and tunicate Ciona intestinalis. Our analyses show that the CYP1 line arose early in deuterostome evolution, with definite CYP1 homologs present in tunicates but more distantly related sequences present in sea urchins. We are combining these sequence-level analyses with ensemble homology modeling and ligand docking to evaluate aspects of CYP1A function, including metabolite formation and uncoupling. Our docking results show that PHAH bind significantly closer to the heme in mammalian CYP1A1 than in fish CYP1A. This result is consistent with experimental evidence that TCB and TCDD are metabolized at least two orders of magnitude faster by mammalian than by fish CYP1As. Furthermore, ensemble modeling explains the PAH metabolite profile differences observed between fish and mammals. These types of computational molecular level analyses of PHAH and PAH ligand interaction with different CYP enzymes thus may allow the prediction of interspecies metabolite and toxicity differences. (NIH grants 2-P42-ES07381, 5-F32-ES012794, GM64700, and EPA R827102-01-0)

Contact: Jared Goldstone Redfield 3-52 MS#32 Woods Hole Oceanographic Institution Woods Hole, MA 02543 Email: jgoldstone@whoi.edu

NEUTRAL MOLECULAR MARKERS AND THEIR VALUE IN ASSESSING IMPACTS TO WILDLIFE POPULATIONS.  Amy M. McMillan (MCMILLIAM@buffalostate.edu), Buffalo State College, Buffalo, NY 14222.

Neutral molecular markers represent areas of the genome with no apparent functional significance. Two useful applications of these kinds of markers for ecological risk assessment are quantifying the amount of genetic variation within populations and defining connectivity between populations. Genetic variation within populations has been used to indicate population health. Populations with low genetic diversity are assumed to be negatively impacted and at risk whereas populations with high genetic diversity are assumed to be robust and evolutionarily stable. Population connectivity reflects the movement of genes between populations. High connectivity is assumed to help rescue impacted populations with an influx of new genes whereas populations with low or no connectivity may not recover. Two examples will be discussed where genotyping organisms has provided insights into population responses to ecological impacts. The first involved an analysis of AFLP (amplified fragment length polymorphism) markers to estimate inter-population relatedness and intra-population genetic diversity in fish exposed or not exposed to contaminants. In the second study, polymorphic microsatellite loci were used as indicators of disease impacts in migratory bird populations. I will address the importance of experimental design in these kinds of studies as well as the benefits and limits to using these and other genotyping methods to assist in ecological risk assessment.

Contact: Amy M. McMillan Biology Department, Buffalo State College 1300 Elmwood Ave., Buffalo, NY 14222 Email: mcmillam@buffalostate.edu

*THE LITTLE SKATE (LEUCORAJA ERINACEA) AS AN IN SITU INDICATOR OF ENVIRONMENTAL SPERMATOTOXICANTS. K.B. Engel (kabonner@bu.edu), G.V. Callard, GV, Boston University, Boston, MA 02215; R. Merson, M.E. Hahn, Woods Hole Oceanographic Institute, Woods Hole, MA 02543.

Epidemiological and wildlife studies have implicated environmental endocrine disrupting chemicals (EDC) in declining sperm counts and gonadal abnormalities. To detect and localize spermatotoxic pollutants in the marine environment without a priori knowledge of their chemical nature, concentration, or mechanism of effect, we are developing the little skate as a sentinel. Skates are nonmigratory and possess a testicular organization that facilitates stage-by-stage analysis of spermatogenesis. Skates were collected from 3 polluted sites, differing in proximity to the highly polluted Superfund site (New Bedford Harbor MA; NBH), and from 5 reference sites (ME, NH, MA). Histological analysis showed that NBH when compared to reference skates had reduced testis size and fewer spermatocysts (germ cell/Sertoli cell clones) in the spermatocyte and early spermatid stages of development, suggesting a defect in the advance of spermatogonial clones into meiosis. Consistent with high levels of PCBs known to be present in NBH, P4501A mRNA was induced in the testis of NBH skates. Also, testicular aromatase mRNA was elevated ~2-fold in NBH. Moreover, skates from polluted sites displayed a gradient of effect for all parameters examined, correlating to their proximity to NBH. We conclude that skates are reliable indicators of the point source and topography of spermatotoxic pollutants. Supported by an MIT Sea Grant and EPA STAR Grant (R825434).

Contact: Kelly Bonner Engel
Boston University, Biology Department
5 Cummington St., Boston, MA 02215
Email: kabonner@bu.edu

POSTER SESSIONS  &  RECEPTION

FUGAWEB, A PROBABILISTIC FUGACITY AND FOOD WEB MODEL FOR ASSESSING ECOLOGICAL RISKS ASSOCIATED WITH CONSUMER PRODUCTS.  M.J. Bock (mbock@Environcorp.com), ENVIRON International Corporation, Portland, ME 04101; and T.R. Barber and M. Ferguson, ENVIRON International Corporation, Burton, OH  44021.

A model was developed to predict the environmental fate and potential for ecological toxicity of new chemicals in consumer products. The model is based on the removal efficiency of wastewater treatment plants coupled to a Tier III Fugacity Model (Mackay et al., 1996). The outputs of the fugacity compartment are used as inputs into a bioaccumulation compartment which is based on Aquaweb (Arnot and Gobas, 2004). Finally, the resulting media specific chemical concentrations and tissue concentrations are used as inputs to a concentration- and dose-based ecological exposure assessment. The media specific concentrations, biota tissue concentrations, and estimated doses are used in the assessment of the ecological effects of the target chemical on various ecological receptors. The inputs are modeled probabilistically to the extent practical (e.g., watershed area; watershed population; sediment depth; organic carbon content, dietary preferences, etc.). The input parameters are designed to span the range of likely values occurring in the continental United States, rather than explicitly account for uncertainty. The results of the model are used to predict the potential for ecological impacts associated with the ingredient of consumer product.

Contact: M.J. Bock, ENVIRON International Corporation, Portland, ME 04101

*Non-Lethal effects due to pesticides’ exposure.  Kathryn Dunphy¹ (kathryn@dal.ca), Laurie Tremblay², Jim Leonard³ and Jocelyne Hellou ^{3,4.  1}Department of Biology, Dalhousie University, Halifax, Nova Scotia, Canada; ²Department of Earth Sciences, Dalhousie University, Halifax, Nova Scotia, Canada; ³Bedford Institute of Oceanography, Department of Fisheries and Oceans, Dartmouth, Nova Scotia, Canada; and ⁴Chemistry and Oceanography Departments, Dalhousie University, Halifax, Nova Scotia, Canada.

More than twenty insecticides, fungicides or herbicides are used to prevent damage to crop grown on Prince Edward Island. Pesticides have also been linked to fish kills witnessed in PEI rivers after rainfall events. Unlike the presence of moribund fish, sub-lethal effects on aquatic organisms are more difficult to assess. Our laboratory studies aimed to determine if invertebrates commonly found in estuaries and reported as tolerant to variable levels of salinity could be affected by pesticide runoff. Six pesticides, namely atrazine, azinphos-methyl, carbofuran, chlorothalonil, endosulfan and hexazinone were used over a gradient of concentrations added to water or sediments. As well, the pesticides partitioning between reference sediments and water was measured. Behavioural experiments were undertaken with the amphipod Corophium volutator and the snail, Ilyanassa absoluta. Concentrations eliciting an avoidance response to spiked compared to reference sediments were observed in amphipods at lower concentrations of hexazinone < chlorothalonyl < azinphos-methyl < endosulfan < atrazine, but not for carbofuran. Concentrations leading to an avoidance of water were determined with snails, while flipping times or attraction to food did not represent sensitive end points. A high amount of distress was determined in snails exposed to Chlorothalonil at 10, 100 and 1000ng/mL and Carbofuran at 100 and 1000ng/mL.

Contact: Kathryn Dunphy, Department of Biology, Dalhousie University, Halifax, Nova Scotia, Canada.

THE CYTOTOXICITY AND GENOTOXICITY OF HEXAVALENT CHROMIUM AND HYPOXIA IN MEDAKA CELLS.  Britton C. Goodale¹ (britton.goodale@maine.edu), Sandra S. Wise¹, Ronald B. Walter², Zehnlin Ju² and John Pierce Wise Sr¹.  ¹Wise Laboratory of Environmental and Genetic Toxicology, Maine Center for Toxicology and Environmental Health, University of Southern Maine, Portland, ME. ²Texas State University, San Marcos, TX.

Marine pollution poses a significant ecological and human health problem. Chromium is a common sediment pollutant that is discharged from metal-finishing, leather tanning and textile dyeing industries. Hypoxia (dissolved oxygen below 2.8 mg O2/L) is caused by increased nutrient levels and temperature in water with poor circulation, and often results from agricultural runoff. Hypoxia has significant physiological effects on marine organisms and is associated with mass mortality events. Medaka is a well-established model organism for marine ecosystem toxicological research. We utilized cells derived from the fin of adult Medaka to study the cytotoxicity and genotoxicity of hexavalent chromium (Cr(VI)) and hypoxia in fish. We found that Cr(VI) was cytotoxic to medaka cells as 24 h treatments of 25, 50, and 100 uM sodium chromate caused a concentration-dependent decrease in relative survival of 31, 19 and 3 percent respectively. Cr(VI) was also genotoxic causing chromatid exchanges and lesions. Hypoxia was not cytotoxic as 21% O2 (normoxia), 8% O2 (physiologic normoxia) and 2% O2 (low oxygen) showed no significant differences after 24 or 120 h exposures. This work was supported by subaward Agreement no. 8000000281.1 (J.P.W.) and Grant number NA04NOS4260202 from the US Department of Commerce, National Oceanic and Atmospheric Administration (R.B.W.).

Contact: Britton C. Goodale, Wise Laboratory of Environmental and Genetic Toxicology, Maine Center for Toxicology and Environmental Health, University of Southern Maine, Portland, ME.

INTEGRATING ECOLOGICAL RISK ASSESSMENT AND NATURAL RESOURCE DAMAGE ASSESSMENT: A HYPOTHETICAL CASE STUDY INVOLVING MULTIPLE RESOURCES.  Allison Guinan (AGuinan@entrix.com) and H. Byrd (HByrd@entrix.com) ENTRIX, Inc, Methuen, MA 01844; and ENTRIX, Inc. New Castle, DE 19720.

Federal and State trustees have become increasingly active in pursuing Natural Resource Damage Assessments (NRDA) for multiple resources, including aquatic and terrestrial habitats, ground water, wildlife, and human uses. Ecological risk assessment (ERA) is part of established methods for site investigation and remediation. Integrating NRD considerations into risk assessment and remedial decision processes has several advantages, including opportunities for shared data collection and analysis, integrated remediation and restoration projects, and potential cost savings. One challenge of integrating ERA and NRDA include establishing relationships between service losses for different resources used in NRDA and the hazard indices from the risk assessment.

This poster highlights the advantages and challenges of integrating ecological risk assessment and NRDA in the remedial process through real-world experiences. A hypothetical case study of an industrial site adjacent to an estuarine wetland is used to demonstrate the complexities of the NRDA process in today’s regulatory atmosphere.

Contact: A. Guinan, ENTRIX, Inc, Methuen, MA 01844

ENTERIC ADENOCARCINOMA IN THE EASTERN OYSTER, CRASSOSTREA VIRGINICA.  Doranne Borsay Horowitz (borsay.dodi@epa.gov), Atlantic Ecology Division, US EPA, NHEERL, 27 Tarzwell Drive, Narragansett, RI 02882 and Inke Sunila (isunila@snet.net), CT Dept of Agriculture, Milford, CT.

In the routine screening of oysters bound for market by the CT Department of Agriculture, several cases of enteric adenocarcinoma were discovered in eastern oysters, Crassostrea virginica, collected from areas along the Connecticut coastline in Long Island Sound. While disseminated neoplasia in bivalves is relatively common and may occur at epizootic prevalences, solid tumors in bivalves are extremely rare. Prevalence was 13 out of 10,801 (0.1%) oysters surveyed between 1997 and 2004 in Connecticut. Affected oysters were found in samples collected from different sites in coastal Connecticut such as the Pawcatuck River, Black Rock Harbor, the Housatonic River seed beds, and the Branford, Milford and Norwalk growing areas. Similar adenocarcinomas have been previously described by the EPA’s Atlantic Ecology Division (AED) from Pawcatuck River and Black Rock Harbor studies the 1980’s. A comparison of oysters collected in studies by CT Dept of Agriculture and the AED was done to further describe the extent of this lesion in the population. In this study thousands of animals from Maine through the Chesapeake Bay region were screened for these enteric lesions from the pathology collections at AED of field collected oysters from 1970 through early 1990’s. No similar neoplasia was detected at other sampling sites. Tumors consisted of single or multifocal basophilic lesions affecting areas of enteric epithelium from the anal to the stomach regions. Lesions were well-differentiated with neoplastic, mitotically-active cells. Neoplastic cells formed glandular structures, the lumina of which contained hemocytes or PAS-positive mucin. In most cases lesions did not penetrate basement membranes and were classified as adenocarcinoma in situ. While the etiology of this tumor is unknown, its occurrence in specific populations might prove significant to future research. Several cases were submitted to the Registry of Tumors in Lower Animals (RTLA 6549-6551).

Contact: Doranne Borsay Horowitz, Atlantic Ecology Division, US EPA, NHEERL, 27 Tarzwell Drive, Narragansett, RI 02882, Email: borsay.dodi@epa.gov

*semiconductor Nanoparticle Uptake and Toxicity correlates with Particle Size and core degradation.  A.N. Johnson (adam.johnson@umit.maine.edu), G. D. Mayer, University of Maine, Department of Biochemistry, Microbiology and Molecular Biology, Orono, ME 04469 and B.E. Bragdon, A. Nohe, University of Maine, Department of Chemical Engineering, Orono, ME 04469.

The rapidly expanding field of nanotechnology has developed exciting new tools beneficial for research. Use of these in vivo products may prove to be less invasive to the patient and also more specific than former diagnostics. Utilization of quantum dots is becoming a popular method of cell-specific imaging due to their fluorescent properties and ease of conjugation to macromolecules. However, the use of quantum dots in humans is questionable due to the composition of the quantum dot core. Since most types of semiconductor nanoparticles are comprised of a cadmium-selenide semiconductor composite, degradation and release of free cadmium is of great concern. Like many molecules of this size, nanoparticles can easily penetrate exterior barriers of organisms and potentially enter cell membranes to exert adverse effects. This also becomes more relevant on a global scale due to disposal of many electronic devices that have recently begun utilizing semiconductor nanoparticles. To examine these concerns, we exposed both transgenic zebrafish embryos and cultured liver cells to various sized quantum dots. Use of confocal microscopy and quantitative fluorescence assays have been employed to assess the nature of quantum dot uptake and cellular response.

Contact: Adam Johnson, 5735 Hitchner Hall, Orono, ME 04469.  Email: adam.johnson@umit.maine.edu

*CYP1A1 EXPRESSION IN SPERM WHALE (PHYSETER MACROCEPHALUS) SKIN BIOPSIES FROM THE PACIFIC OCEAN SHOWS SITE BUT NOT SEX DIFFERENCES. Carolyne LaCerte^1,2 (Carolyne_Lacerte@umit.maine.edu), Rebecca Clark², Ildiko Polyak², John Cannon², Kim Pinkerton², Nadia Rubio-Cisneros³, Sarah Mesnick⁴, Michael Moore5, Roger Payne², John P. Wise Sr.¹, John J. Stegeman5, and Céline A.J. Godard-Codding^1,2. ¹Wise Laboratory of Environmental and Genetic Toxicology, University of Southern Maine, Portland, ME 04104, USA; ²Ocean Alliance, Lincoln, MA 01773, USA; ³Centro de Investigationes Biologicas del Noroestre (CIBNOR), La Paz, B.C.S., Mexico; ⁴Southwest Fisheries Science Center, NOAA, La Jolla, CA 92037, USA; 5Woods Hole Oceanographic Institution, Woods Hole, MA 02543, USA.

The cytochrome P401A1 (CYP1A1) enzyme can metabolize and/or activate environmental pollutants, such as polychlorinated biphenyls and polycyclic aromatic hydrocarbons. In many animal species CYP1A1 induction is used as a biomarker of exposure to the above contaminants and we recently validated its use in cetaceans. CYP1A1 induction was examined using immunohistochemistry in 235 sperm whale skin biopsies collected from five regions within the Pacific Ocean (Sea of Cortez (78), Galapagos (25), Pacific Crossing (35), Kiribati (17) and Papua New Guinea (80)). CYP1A1 expression was measured in endothelial, smooth muscle and fibroblast cells. CYP1A1 was expressed in all three cell types but was highest in the smooth muscle cells. Our data shows no significant difference in CYP1A1 induction between males and females, but a significant difference among sites. CYP1A1 induction was highest in the Galapagos samples, high in the Sea of Cortez and relatively low in the three other sites. The high CYP1A1 induction levels found in the Galapagos samples suggest higher exposure to environmental pollutants in whales from this region, therefore warranting further investigation into possible health effects and pollution sources.

Contact: Carolyne LaCerte, Wise Laboratory of Environmental and Genetic Toxicology, University of Southern Maine, Portland, ME 04104 and Ocean Alliance, Lincoln, MA 01773.

INTEGRATION OF ARCVIEW AND EVS FOR THREE-DIMENSIONAL MODELING OF SEDIMENT DEPTH DATA FOR A CREEK IN SOUTHWESTERN OHIO.  Erik Martin (emartin@environcorp.com), ENVIRON International Corp., Portland, ME, Tim Barber, Ph.D. and Michael Ferguson ENVIRON International Corp., Burton, OH, and Angela DeDolph, ENVIRON International Corp., Deerfield, IL.

Accurate, simple and understandable three-dimensional computer models are increasingly important for planning remediation of contaminated stream sediments. Model imagery not only reveals patterns and makes data more accessible to a wider audience, but it also helps guide field remediation efforts. In order to accurately model the sediment profile and associated contamination within an Ohio creek, water and sediment depths were probed at regular intervals along a two mile reach in conjunction with a sediment sampling plan. Measurements were recorded in river feet relative to a known datum and were converted into X and Y coordinates in ESRI’s ArcView 9.1 GIS. These data, along with surveyed elevation transects, were exported to C-Tech’s Environmental Visualization System (EVS), which krigged the data and created a three-dimensional representation of the creek. The three-dimensional image could be sliced at any point along the length of the modeled creek to view interpolated cross-sections. The model, in concert with sample concentration data, allows calculation of the volume of contaminated sediment, so that field crews can be directed to excavate to the appropriate elevation at any given location. This degree of precision helps ensure that contaminated sediments are removed to the specified remediation goal, while minimizing disturbance of uncontaminated sediment. In order to further improve accuracy, future plans include the integration of digital topographic lines to supplement surveyed elevation transects, thereby reducing the dependence on interpolated elevation data.

Contact: Erik Martin, ENVIRON International Corp., Portland, ME.

*EVALUATION OF THE EFFECTS OF SEDIMENT CONTAMINANT RELEASE.  Danielle M. Miniutti (danielle.miniutti@umit.maine.edu), Emily G. Notch, and Gregory D. Mayer, Biochemistry, Microbiology, & Molecular Biology Department, University of Maine Orono.

One factor that is known to contribute to the decline of Atlantic salmon (Salmo salar) populations is lack of access to quality spawning areas upstream of dams. Two dams that hinder salmon migration have been slated for removal from the Penobscot River near Bangor, Maine in an effort to revive endangered Atlantic salmon populations as part of the Penobscot River Restoration Project. Due to mechanical perturbation of sediment stores behind dams, their removal can result in release of sediment bound toxicants into the river water column. This potential dissolution of toxicants could presumably increase bioavailability of sediment-bound toxicants and induce deleterious effects in aquatic species associated with the perturbed water column. In an effort to characterize the toxicity of resuspended sediments from the two dams, multiple biomarkers were analyzed to examine effects of resuspended contaminants on aquatic organisms. The presence of estrogen receptor agonists and aryl hydrocarbon receptor agonists in the resuspended water was analyzed utilizing estrogen- and Ah-induced, cell based, transcriptional assays incorporating luciferase as a reporter gene. Furthermore, adult and embryonic zebrafish (Danio rerio) were exposed to resuspended water and analyzed for transcription of vitellogenin, metallotheionein, and cyp1A1 using fluorescence based qRT-PCR. These three biomarkers are indicative of estrogens, heavy metals, and polycyclic aromatic hydrocarbons, respectively. Analysis of potential toxicant release prior to dam removal will help direct remediation efforts within the overall restoration plans.

Danielle M. Miniutti 5735 Hitchner Hall, University of Maine, Orono Maine 04469.  Email: danielle.miniutti@umit.maine.edu

*ESTROGENIC COMPOUNDS IN HALIFAX HARBOUR:  ANALYTICAL METHODS OF DETECTION AND ENVIRONMENTAL FATE STUDIES.  B. Robinson, Dept. of Oceanography, Dalhousie University, Halifax, NS; and J. Hellou, Dept. of Fisheries and Oceans, Bedford Institute of Oceanography, Dartmouth, NS.

There is growing concern over the occurrence of natural and synthetic estrogens in waterways, since low concentrations of endocrine-disrupting compounds (EDC’s) have been linked to reproductive and developmental effects in aquatic organisms. The occurrence and especially partitioning of EDC’s has received less attention in marine relative to freshwater environments. An analytical method was developed to detect for the phenolic targets 17β-estradiol (E2), ethynylestradiol (EE2) and bisphenol-A (BPA) in samples of seawater and sediment. The compounds were extracted from seawater using solid phase extraction and from sediment using a sonic probe. Extracts were fractionated to help remove less and more polar anthropogenic chemicals, prior to the identification and quantification using negative-ion LC-MS/MS operating in MRM mode. Results from the analysis of sites in the harbour revealed low concentrations (ng/L and ng/g) of all three target compounds over one year of sampling. To study the partitioning and biodegradation of EDC’s in marine samples, seawater/sediment mixtures from the harbour were spiked with the three target compounds and concentrations in the aqueous and sediment phases were monitored for up to 28 days. The following ranking of biodegradation rates E2 > BPA > EE2 was consistently observed, but variability in degradation rates did not always correlate with measured microbiology parameters. This indicates that the chemical degradation may be more important than biological degradation for these EDC’s, although microorganisms may still play a key role. In general, partitioning was directly related to the organic carbon content of the sediment, with the greatest levels of partitioning for EE2 and least for BPA.

Contact: B. Robinson, Dept. of Oceanography, Dalhousie University, Halifax, NS

*THE CYTOTOXICITY AND GENOTOXICITY OF CR(VI) IN LUNG AND TESTES CELLS FROM THE NORTHERN RIGHT WHALE (EUBALAENA GLACIALIS).  Fariba Shaffiey¹ (fariba.shaffiey@maine.edu), Sandra S. Wise¹, Scott Kraus², Todd O’Hara³ and John Pierce Wise, Sr.¹; ¹Wise Laboratory of Environmental and Genetic Toxicology, Maine Center for Toxicology and Environmental Health, University of Southern Maine, 96 Falmouth St., Portland, ME. 04104; ²New England Aquarium, Edgerton Research Laboratory, Central Wharf, Boston, MA 02110; ³Institute of Arctic Biology, University of Alaska, Fairbanks, AK 99775.

The Northern Right Whale was hunted to the brink of extinction. Despite international protection since 1935, population recovery has been stagnant and in decline. Only about 300 individuals remain. Our research investigates whether environmental contaminants, specifically hexavalent chromium (Cr(VI)), induce cytotoxicity and genotoxicity in cells from reproductive organs of this endangered species. We found that Cr(VI) induced time- and concentration-dependent cytotoxicity in right whale testes and lung cells. In lung cells, after a 24 h exposure to 2.5 and 10 uM sodium chromate, the relative cell survival was 74 and 32 percent respectively, which decreased to 16 and 0 percent at these respective concentrations after a 120 h exposure. In testes cells, the relative survival after 2.5 and 10 uM was 63 and 2 percent respectively, which decreased to 28 and 0 percent at these respective concentrations after a 120 h exposure. Cr(VI) was also genotoxic. In lung cells, 2.5 and 10 uM damaged 13 and 43 percent of metaphases after a 24 h exposure respectively. In testes cells, these concentrations induced 13 and 34 percent of metaphases respectively. This work was supported by grant number NA03NMF4720478 from the United States Department of Commerce, National Oceanic and Atmospheric Administration (J.P.W.).

Contact: Fariba Shaffiey Wise Laboratory of Environmental and Genetic Toxicology Maine Center for Toxicology and Environmental Health University of Southern Maine, 96 Falmouth St., Portland, ME 04104

*FATE OF ESTROGENIC COMPOUNDS IN THE MUD SNAIL.  A. Parsons^1,2 (ashley.parsons@dal.ca), K. Campbell^1,2 (campbell_katie@hotmail.com) and J. Hellou^1,2 (HellouJ@mar.dfo-mpo.gc.ca), ¹Chemistry Department, Dalhousie University, Halifax, NS, B3H 4J3; ²Bedford Institute of Oceanography, 1 Challenger Drive, Dartmouth, NS, B2Y 4A2.

Some xenobiotics such as pharmaceuticals can mimic the endogenous estrogen, estradiol, and their uptake may lead to detrimental effects within aquatic life. The abundant priority pollutants, polycyclic aromatic hydrocarbons (PAHs), are also potentially estrogenic due to their reactivity within organisms. Large snails can be found in the offshore and represent an attractive species for the fishing industry. Many other species of snails, including the mud snail, Ilyanassa obsoleta, have an inter-tidal habitat and can be found in harbours, where they can be exposed to contaminants present in water, food, or sediment deriving from sewage effluents. In our lab, the offshore whelk, Buccinum undatum, were found to bioaccumulate and biotransform a model PAH, pyrene. The ability of the nearly one hundred times smaller mud snails to bioaccumulate and biotransform anthropogenic compounds was also investigated. Results regarding the choice of tissue parts, lipid content, and approach for contaminants' analyses including quality assurance/ quality control protocol will be presented. Recoveries of three surrogate standards representative of a wide range of polarities and analysed by HPLC with ultra-violet and fluorescence detectors will be outlined.

Contact: Ashley Parsons, Chemistry Department, Dalhousie University, Halifax, NS, B3H 4J3.  Email: ashley.parsons@dal.ca

*THE EFFECTS OF PCBs ON IMMUNE FUNCTIONS OF THE EASTERN OYSTER.  J. M. DiGalbo, S. De Guise, Department of Pathobiology and Veterinary Science, University of Connecticut, Storrs, CT 06269.

PCBs have been shown to have toxic effects on immune functions of several laboratory and wild animals. The effects that PCBs have on the immune functions of oysters are not known. This study looks at the effects PCBs have on the immune functions of the Eastern Oyster relevant to resistance to the parasite Perkinsus marinus, specifically phagocytosis. Hemolymph was collected from the oyster and hemocytes were incubated in vitro with PCBs for three hours. The PCBs tested include the coplanar PCB 169 and the non-coplanar PCBs 138, 153, and 180 at concentrations ranging from 0 ppm to 20 ppm. The hemocytes were then incubated with latex beads for one hour. Flow cytometry was used to determine how many cells had ingested one or more beads. The four PCBs tested did not modulate the phagocytosis of latex beads by the hemocytes. Though PCBs did not affect the phagocytosis of the latex beads, further studies are being done to determine if PCBs modulate the phagocytosis of the parasite Perkinsus marinus, and whether the ingestion of the parasite further modulates the phagocytosis of latex beads, as well as the modulation of hemocyte apoptosis upon infection, all of which were previously determined as important in disease resistance.

Contact: J. M. DiGalbo Department of Pathobiology and Veterinary Science, University of Connecticut, Storrs, CT 06269

*In Vitro effects of individual polychlorinated biphenyl congeners on lymphocyte proliferation in HUMANS, MARINE MAMMALS AND MICE.   H. Leibrecht, C. Mori, M. Levin, M. Trailsmith, S. De Guise, Department of Pathobiology and Veterinary Science, University of Connecticut, Storrs, CT 06269.

There is both epidemiological and experimental evidence which suggests PCBs cause immunomodulation. Non-ortho (coplanar) PCBs are structurally similar to TCDD and as such mediate their effects via the aryl hydrocarbon receptor (AhR). Non-coplanar PCBs, on the other hand, have been shown to produce AhR-independent immunomodulatory effects. The aim of this study was to assess if PCB structure could predict PCB-induced changes in lymphocyte proliferation. Lymphocytes of mice, humans and marine mammals were exposed in vitro to increasing concentrations of non-ortho (81, 126, 169), mono-ortho (156, 157) or di-ortho (153, 138, 180) PCBs and stimulated to proliferate with the mitogens Concanavalin A (ConA) or lipopolysaccharide (LPS). Dose response curves were generated and compared within and across species. Results showed that PCB congeners of the same class do not always produce the same effect within species. For example, PCB 81 produced a significant increase of ConA induced mouse lymphocyte proliferation whereas PCB 126 produced a significant decrease. In addition, individual PCB congeners induced distinctly different changes in different species. Upon exposure to PCB 156, ConA-induced lymphocyte proliferation was significantly decreased in mice but increased in humans. The effects of PCBs can not be predicted by structural class nor can the effects in one species predict the effects in others. Therefore, risk assessments could be improved through the use of species-specific and congener-specific data, such as that produced in this study.

Contact: H. Leibrecht Department of Pathobiology and Veterinary Science, University of Connecticut, Storrs, CT 06269

IMPACTS OF MULTIPLE STRESSORS ON COMMON LOONS IN NEW HAMPSHIRE, USA: A DEMONSTRATION STUDY FOR STRESSOR EFFECTS ACROSS SPACE.  Steven Walters¹, Anne Kuhn¹ (kuhn.anne@epa.gov), Jason S. Grear¹, Matthew C. Nicholson², Jane Copeland³, Steven A. Rego¹, and Diane E. Nacci¹.  ¹U.S. Environmental Protection Agency, Office of Research and Development, National Health and Environmental Effects Research Laboratory, Atlantic Ecology Division, Narragansett, RI; ²U.S. Environmental Protection Agency, Region 3, Philadelphia, PA; ³Computer Sciences Corporation.

Factors that significantly impact wildlife population dynamics, such as resource availability and exposure to stressors, frequently vary over space and thereby contribute to the heterogeneous spatial distributions of organisms. The spatial co-occurrence of organisms, environmental factors, and multiple stressors has important implications for ecological risk assessment and conservation management strategies. We are evaluating modeling tools for predicting population responses to multiple stressors in a spatially explicit framework. The goal of the present study is to illustrate how localized effects of spatially heterogeneous stressor distributions can impact the regional persistence or decline of wildlife populations. Using the modeling package RAMAS-GIS, we explored the impact of spatial dependence in organism and stressor distributions, using common loon (Gavia immer) populations in New Hampshire and exposure to chemical and land use stressors as a case study. Environmental data and lake-specific loon demographic information, population and habitat suitability models, and estimates of dispersal behavior were integrated in RAMAS to simulate localized population and regional metapopulation trends in NH under hypothetical unstressed conditions. These results were then compared with dynamics under simulated stressor-induced decreases in fecundity on source versus sink lakes. Decreases in fecundity on source lakes resulted in metapopulation occupancy rates and probabilities of persistence that were lower than those observed under unstressed conditions. However, comparable decreases in fecundity on sink lakes did not appreciably alter dynamics relative to those observed under unstressed conditions: only when accompanied by reductions in dispersal rates did impairment within sinks result in lower occupancy rates. The results illustrate the importance of accounting for spatial heterogeneity in population and stressor distributions, as well as understanding the degree to which dispersal connects localized populations, when assessing ecological risks from multiple interacting stressors. Further analyses will be used to explore and test effects of stressors of concern, such as lake acidification and increases in human development.

Contact: Anne Kuhn, U.S. Environmental Protection Agency, Office of Research and Development, National Health and Environmental Effects Research Laboratory, Atlantic Ecology Division, Narragansett, RI.  Email:  kuhn.anne@epa.gov

*CYTOTOXICITY AND GENOTOXICITY OF MERCURY IN BOWHEAD WHALE (BAELAENA MYSTICETUS) AND HUMAN LUNG CELLS.  Heather Throckmorton¹ (Heather.throckmorton@maine.edu), Sandra S. Wise¹, Rebecca Kelly¹, Scott Kraus², Todd O’Hara³, Tracy Romano⁴, John Pierce Wise, Sr.¹; ¹Wise Laboratory of Environmental and Genetic Toxicology, University of Southern Maine, 96 Falmouth St., Portland, ME 041042 New England Aquarium, ²Edgerton Research Laboratory, Central Wharf, Boston, MA 021103 Institute of Arctic Biology, ³University of Alaska, Fairbanks, AK 99775, ⁴Mystic Aquarium, 55 Coogan Blvd, Mystic, CT 06355.

Mercury is persistent in the environment and has recently become a widespread human health concern. This study examines the cytotoxic and genotoxic effects of mercury chloride in human and bowhead whale lung cells. Preliminary data indicate that mercury begins to cause cytotoxic effects for both cell types at 50 uM. However, mercury did not induce chromosome damage at any concentration tested. We considered genotoxicity on a finer scale by using an immunofluorescence staining technique to identify gamma-H2AX, a protein responsible for the repair of DNA double strand breaks. This protein is phosphorylated specifically in response to DNA double strand breaks and therefore the presence of gamma-H2AX within the nucleus of a mammalian cell represents a clear indication of double strand breaks. We found that mercury does induce gamma-H2A.X foci formation in bowhead whale lung cells thus indicating that mercury is genotoxic to whale cells. This work was supported by Grant number NA03NMF4720478 from the United States Department of Commerce, National Oceanic and Atmospheric Administration (J.P.W.).

Contact: Heather Throckmorton 37 Merrill St., Portland, ME 04101.  Email: heather.throckmorton@maine.edu

*GENE EXPRESSION IN ZEBRAFISH EMBRYOS AS AN ENDOCRINE DISRUPTOR SCREEN. Lucinda Burnam (lcburnam@bu.edu), Apolonia Novillo, and Gloria V Callard, Boston University Department of Biology, Boston MA, 02215.

By virtue of their ability to bind to hormone receptors, chemicals of diverse structure and origin have been reported to interfere with normal development and reproduction (termed endocrine disrupting chemicals (EDC): however, few of the ~87,000 chemicals added to the environment have actually been screened for EDC effects. Here, we test the hypothesis that gene expression in zebrafish embryos can be used for screening EDC without a priori knowledge of their chemical nature, concentration, uptake, access to targets, receptor binding, or mechanism of action. Initial studies selected genes known to be markers of estrogen receptor (ER) or arylhydrocarbon receptor (AhR) signaling, and to represent different tissues types and cellular functions. Real-time quantitative (q)RT-PCR assays were developed for measuring multiple different mRNAs from a given sample. Using authentic ER or AhR ligands (estradiol, dioxin), treatment protocols were optimized for dose, duration and stage-of-development. Dose-response characteristics were gene-, ligand- and stage-specific. Using a single optimized protocol, testing was expanded to include diverse ER and AhR ligands, alone and in combination; and chemicals that act upstream or downstream of receptor binding. We conclude that zebrafish embryos have the convenience of an in vitro assay with the added value of a whole animal endocrine system for identifying EDC. Supported by an EPA STAR grant (RD831301) and an EPA STAR Fellowship (FP91653101).

Contact: Lucinda Burnam, 5 Cummington St., Boston, MA 02215  Email: lcburnam@bu.edu

* denotes student presentations.