SDMS DocID 2009094

Evaluation of Liver and Skin Tumor Prevalence in Fish from theDelaware Estuary Watershed

Proposed by

Fred Pinkney, Ph.D.Senior Scientist

Environmental Contaminants ProgramU.S. Fish and Wildlife ServiceChesapeake Bay Field Office177 Admiral Cochrane Drive

Annapolis, MD 21401tel: 410-573-4519fax: 410-269-0832

Fred Pinkney@fws.gov

for

David CarterDelaware Coastal Management Program

Delaware Department of Natural Resources and Environmental Control89 Kings Highway, P.O. Box 1401

Dover, DE 19903

Mav 2003

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Project Title: Evaluation of Liver and Skin Tumor Prevalence in Fish from the DelawareEstuary WatershedPrincipal Investigator: Fred Pinkney, Ph.D., U.S. Fish and Wildlife Service, Chesapeake BayField Office, Annapolis, MD. Tel: 410-573-4519, Fax: 410-269-0832Collaborator: John Harshbarger, Ph.D., George Washington University Medical Center,Washington, DC, Tel. 202-994-1056, Fax: 202-994-2618

INTRODUCTION

Tumor surveys in bottom-dwelling fish have been used widely as an indicator of contaminationand a method for monitoring the success of cleanup actions (e.g., Baumann el al. 1996, Baumannand Harshbarger 1998, Myers et al. 1994). In freshwater ecosystems, the brown bullhead (Ameiurus nehulosus) has been shown to develop skin and liver tumors in response tocontaminant exposure, with the most persuasive linkage for areas with polynuclear aromatichydrocarbon (PAH) -contaminated sediments (Baumann and Harshbarger 1998). Baumann et al.(1996) suggested that, in brown bullheads, liver tumor prevalence above 9% and skin tumorprevalence above 20% were nearly always observed in chemically contaminated areas.Recently, Baumann (2002) refined these estimates and stated that liver tumor prevalence aboveabout 5% and skin tumor prevalence above 12% could be used to distinguish between highlycontaminated and less contaminated areas in the Great Lakes. The U.S. Fish and WildlifeService, Chesapeake Bay Field Office (CBFO) has developed a data base on tumors in bullheadsfrom the tidal freshwater areas of the Chesapeake Bay watershed (Pinkney et al. 1995, 2001,2002).

Brown bullheads have been captured in waters with salinity as high as 15 parts per thousand(Boyer 1995) but they are primarily a freshwater species and can be difficult to locate inestuarine waters. The mummichog, Fiindulus hetcroclitus, has been used in tumor surveys inestuaries of the U.S. east coast. Vogelbein et al. (1990) reported a 33% prevalence of livertumors in mummichogs collected near the Atlantic Wood Treating Superfund Site on theElizabeth River, Virginia, where sediments are contaminated with creosote. A sediment samplemeasured in the study at the site with the 33% tumor prevalence contained 2200 ppm total PAHs.In a pilot study, Harshbarger (2003) diagnosed hepatocellular carcinomas in 30% ofmummichogs collected in Hershey Run, adjacent to the Koppers Wood Treating Superfund Site,near Newport, Delaware. In Hershey Run, total PAH concentrations as high as 13,300 ppm hasbeen reported (Woodward Clyde 1996). If a size minimum of 7.0 cm is applied, the Harshbarger(2003) prevalence is increased to 43%. This report is attached as an appendix to this proposal.

Here we propose an approach for an initial tumor survey of selected sites in the DelawareEstuary. We identify areas as possible sites of interest due to sediment contamination withPAHs and other compounds. We propose to conduct sampling at or near the two DelawareEstuarine Research Reserve sites - Blackbird Creek and the Lower St. Jones River - to establishbaselines for long-term monitoring of these areas.

MATERIALS AND METHODS

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Sites of Interest

Area 1: The Christina River is a tidal freshwater system that flows into the Delaware through ahighly industrialized watershed, containing eight federal and several state Superfund sites. Forexample, the Koppers Superfund site is a former wood treating facility that released PAHs andmetals into Hershey Run, and wetlands adjacent to White Clay Creek and the Christina.Junkyards with automobile parts can be observed along the banks of some stretches of the river.Throughout the watershed, sediments are contaminated with polychlorinated biphenyls (PCBs),polynuclear aromatic hydrocarbons (PAHs), pesticides, and toxic metals (Olinger 1997). A fishtissue advisory is in place (R. Greene, Delaware Department of Natural Resources andEnvironmental Control (DNREC), pers. comm.). Olinger (1997) reported the occurrence ofsediment toxicity and measured carcinogenic PAHs in the 20 ppm (wet weight) range usingimmunoassay screening procedures. Limited sampling conducted as part of the Koppersinvestigation, indicated that a Christina River sediment sample collected near the confluence ofWhite Clay Creek contained a total PAH concentration of 23.530 ppm dry weight (Woodward-Clyde 1996). One location in Hershey Run had a total PAH concentration of 13,300 ppm. Thissection of the Christina River (mid-Christina River) was listed on the Delaware 1998 303(d) listof impaired waters due to PCBs and nutrients.

Mummichogs were collected from Hershey Run for the Harshbarger (2003) study and have beencollected from the tidal Christina (Woodward Clyde 1996). Brown bullheads and mummichogsare included in the species lists for the tidal Christina River compiled by Woodward-ClydeAssociates (1992).

Area 2: The industrialized Delaware City area of the Delaware River is impacted by releasesfrom several operating and former facilities. There are reports of repeated National PollutantDischarge Elimination System (NPDES) violations from the Motiva Refinery and severalSuperfund sites (Standard Chlorine, Tybouts Corner) are located in the nearby Red Lion Creeksubwatershed. NOAA (2001) reported a total sediment PAH concentration of 12.6 ppm in asample collected several miles downriver of the refinery outfall. A fish tissue advisory is inplace for this section of the Delaware River (R. Greene, DNREC, pers.comm.). Red Lion Creekcontains a mixture of contaminants in sediments including chlorinated benzenes from theMetachem (Standard Chlorine) Superfund site and PCBs from unknown sources. This creek isalso covered by a fish advisory.

Brown bullheads were observed in small numbers in Red Lion Creek but may be diff icul t toobtain in sufficient numbers for surveys. Mummichogs are likely to be available from both RedLion Creek and the areas near the Motiva facility.

Area 3: One of the two Delaware National Estuarine Research Reserve sites is the Lower St.Jones River Reserve. The St. Jones River watershed drains a portion of the coastal plain incentral Kent County, DE, including the city of Dover, the surrounding suburbs, industrial areas,agricultural areas and Dover Air Force Base (NERR web site:http://inlet.geol.sc.cdii/DEL/st-joncs-river.html). The river is less than pristine; four National

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Priority List Superfund sites, including the former Dover Gas Light facility that released PAHs,are in the watershed. In 1993, DNREC issued a fish tissue consumption advisory based on PCBconcerns for catfish, carp, white perch, and largemouth bass (DNREC 1999). The upper St.Jones River is included on the 303(d) list for nutrients, pathogens, low dissolved oxygen, andPCBs. The Lower St. Jones River is listed on the state's 1998 303(d) list of impaired waters dueto PCBs, nutrients, and low dissolved oxygen.

Brown bullheads have been identified in preliminary trapping efforts in the St. Jones River nearthe Route 13 Bridge. It is possible that electroshocking would produce sufficient numbers inareas near Dover, Delaware. It is likely that mummichogs could be easily collected, especiallyfrom the lower section of the river. Their abundance at the Reserve was documented on the website: http://www.dnrec.state.de.us/dnrec2000/divisions/soil/dnerr/tmpl023300816.htm.

Area 4: The other Delaware National Estuarine Research Reserve is Blackbird Creek.Blackbird Creek is believed to be relatively unimpacted from local sources of industrialcontaminants (D. Carter, DNREC, pers. comm.), although there is concern about non-pointsource pollution from silviculture and agricultural runoff (DNREC 1999). Preliminary surveys(conducted last fall) and salinity data suggest that mummichogs would be the appropriate speciesfor this river.Brown bullhead are listed on the Reserve web sitehttp://inlot.geol.sc.edu/DEL/hlackhird-creck.html as being found in the freshwater areas.

Sampling Design

We propose to conduct a tumor survey in the two industrial areas (the Christina River and theDelaware City area of the Delaware River) and the two NOAA Estuarine Research Reserve(Blackbird Creek and St. Jones River) sites in Delaware. The proposed study would be the firstsystematic fish tumor survey in Delaware and should give DNREC a useful environmentalindicator for watershed monitoring. The proposed areas and sites are listed in Table 1.

We propose to use the mummichog in an initial survey of these areas. Mummichogs are knownto be highly localized (Whitehead 1995) with summer home range of 36 meters in tidal creeksand somewhat more extensive movements in fall and winter. They have considerable contactwith sediments, feeding on bottom-dwelling invertebrates. Advantages of the mummichog intumor surveys are that they are often easy to obtain large numbers at low cost and they have beenshown to develop liver tumors when exposed to high concentrations of PAHs (Vogelbein et al.1990, Harshbarger 2003). Their response to lower PAH concentrations and othersediment-bound carcinogens is not yet known. Based on available data, we believe thatmummichogs will be available from all six of the proposed collection sites (Table 1).

The number of collection sites depends on the availability of funding (see below). Based onconversations with DNREC, we propose to work together with DNREC personnel to obtainsamples. One approach would be to collect mummichogs from two sites in each of the industrialareas and one site in each of the Estuarine Reserves. Thus, for the Christina area, the possiblesites would be (1 ) Hershey Run near the Koppers site, and (2) either Churchman's Marsh or the

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Christina River near White Clay Creek. For the Delaware City Area, the possible sites would be(1) Red Lion Creek, and (2) the near shore area near the Motiva refinery (Table 1).

If requested, we can provide a separate proposal for investigating tumors in brown bullheads.The advantage of bullheads is that there is an extensive data base on tumor prevalence in boththe Chesapeake Bay and Great Lakes watersheds. Thus, we would be able to compare theprevalence in the Delaware Estuary against proposed criteria of 5% liver tumors and 12% skintumors proposed by Baumann (2002) to distinguish highly contaminated Areas of Concern vs.less contaminated Areas of Recovery. Of the four identified areas, we believe that the mostlikely locations for brown bullheads in sufficient size and number would be the St. Jones Rivernear Dover and sections of the Christina River. The disadvantage of the bullhead would be theexpense and difficulty in collecting sufficient numbers offish of appropriate size (>250 mm).

Objectives

1. To evaluate the prevalence of skin and liver tumors in mummichogs from locations in theDelaware River watershed where polynuclear aromatic hydrocarbon (PAH) and other organicchemical contamination is suspected and from the two Delaware National Estuaries ResearchReserves;2. To evaluate the response of mummichogs in areas of extreme (Hershey Run) and less extreme(Churchman's Marsh) sediment contamination;3. To provide environmental managers with a monitoring tool to evaluate habitat quali ty andmonitor changes in loadings and/or cleanup actions.

Methods/Procedures:

We will collect thirty mummichogs (> 70 mm) from each of the six sites. With the assistance ofDNREC, fish will be collected either with minnow traps, seining, or electroshocking. Samplingefforts wil l be coordinated through David Carter and Bob Scarborough of DNREC. Fish wouldbe held in collection site water at one of the Estuarine Reserve facilities and transported live tothe Chesapeake Bay Field Office for necropsy.

Fish wil l be measured, weighed, euthanized, and necropsied. Visible lesions will be noted andexamples will be photographed. Livers will be excised and weighed and visible skin lesions willbe dissected. The organs will be preserved in 10% buffered neutral formalin, embedded,sectioned at 4 um, and stained with hematoxylin and eosin. Dr. John Harshbarger of GeorgeWashington University Medical Center will perform the histopathology and diagnose thepresence of tumors and preneoplastic lesions. A statistical comparison of lesion prevalence willbe performed using an extension of Fisher's Exact Test (Stokes et al. 1995).

Dissemination of Results

Results wil l be presented as an FWS scientific report including a description of the habitats,compilation of existing contaminant and water quality data, statistical comparison of tumorprevalence, and interpretation of findings. A fact sheet wil l be prepared that can be placed on

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FWS and DNREC web pages. The report will be distributed to appropriate state agencies,interagency groups working in the watershed, and to the public. An effort will be made to publishstudy results in a peer-reviewed journal. This may be dependent on the availability of relevantsediment quality data from the collection sites so that inferences can be made on the degree ofcontaminant exposure. If requested, FWS will collect and analyze sediment samples fromtransects at each collection site. Sediments would be analyzed for PAHs (including alkylatedcompounds), PCBs, organochlorine pesticides, grain size, and total organic carbon. FWS contractlaboratories would be used and quality control/quality assurance would be provided by the FWS'Patuxent Analytical Control Facility.

Relevance to Environmental Managers:

The results of the study would be useful in helping to determine the suitability of the mummichogas a biological monitoring tool. The proposed study would serve as a 2003 baseline; a similarsurvey could be repeated several years hence as a tool to monitor habitat quality. Tumor rates inbottom-dwelling fish have been used as such an indicator in the Great Lakes and Puget Sound forthe identification and monitoring of areas of concern (e.g., Hartig et al. 1990). Development of afish tumor indicator would partially address the need identified in the Delaware Estuary Program's Comprehensive Conservation and Management Plan for data on the acute and chronic effects ofcontaminated sediments on aquatic biota.

Project Time Period: June 2003 - March 2004

June -July 2003: Fish collection and necropsy

July - November 2003: Tissue processing and histopathology

December 2003 - January 2004: Preparation of draft report

Final report to be provided 30 days after receipt of all comments

Table 1. Proposed sampling sitesAreaChristina Hershey Churchman's

Delaware CitySt. JonesBlackbird Creek

Site 1Hershey Run near Koppers

near Motiva facilityLower St. Jones- ReserveBlackbird Creek - Reserve

Site 2Churchman's Marsh or Christinanear White Clay C'reekRed Lion Creek

BUDGET

Histopathology: Dr. John Harshbarger, George Washington University Medical Center

Estimate based on 180 samples (6 locations x 30 fish/location)via purchase order from DNREC based on SI00 per fish to Dr. Harshbarger

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$14,000 ($4000 in kind)

FWS charges

CBFO salary (collection and necropsy@10 days (2 biologists x 5 days@$560/day) $4900 ($700 in kind)

CBFO salary (data analysis and report) $2800 (S1120 in kind)7 days@$560/day

Supplies (traps, nets, jars, travel, reagents) $300

Total FWS Direct Cost $8000

Overhead (18%) $1440

Total to FWS $9440

Total Requested for Proposal (based on 180 fish estimate) $23,440

Note: Dr. Harshbarger would only be paid for the sites that are collected. For example, if due tocollection problems, there are only 3 sites (90 fish), his fee would be $7000. The FWS chargeswould not change.

REFERENCES

Baumann, P.C. 2002. Beneficial use impairment #4: Fish tumors and other deformities. In:Workshop on the delisting criteria on the Detroit River Area of Concern. Proceedings of aworkshop held at the University of Windsor, April 26-27, 2002. Environment Canada,Burlington, Ontario, Canada.

Baumann, P.C. and J.C. Harshbarger. 1998. Long term trends in liver neoplasm epizootics ofbrown bullhead in the Black River, Ohio. Environ. Monit. Assess. 53:213-223.

Baumann, P.C., I.R. Smith, and C.D. Metcalfe. 1996. Linkages between chemical contaminantsand tumors in benthic Great Lakes Fish. J. Great Lakes Res. 22:131-152.

Boyer, M.R. 1995. Catfish. In: L.E. Dove and R.M. Nyman, editors. Living Resources of theDelaware Estuary. The Delaware Estuary Program, pp. 157-166.

Delaware Department of Natural Resources and Environmental Control (DNREC). 1999.Delaware National Estuarine Research Reserve Estuarine Profiles. DNREC, Dover. DE.

Harshbarger, J.C. 2003. Hershey Run Mummichog Histopathology Report. Prepared for U.S.Fish and Wildl i fe Service, Chesapeake Bay Field Office, Annapolis, MD.

Hartig, J.H., D.E. Rathke, and D.J. Williams. 1990. How clean is clean in Great Lakes areas ofconcern? J. Great Lakes Res. 16:169-179.

Myers, M.S., C.M. Stehr, O.P. Olson, L.L Johnson, B.B. McCain, S-L. Chan, and U. Varanasi.1994. Relationships between toxicopathic hepatic lesions and exposure to chemicalcontaminants in English sole (Pleuoronectes vetulus), starry flounder (Platichthvs stellatus),and white croaker (Genyonemus lineatus) from selected marine sites on the Pacific coast,USA. Environ. Health Perspect. 102:2-17.

National Oceanographic and Atmospheric Administration (NOAA). 2001. Magnitude andextent of contaminated sediment and toxicity in Delaware Bay. NOS NCCOS CCMA 148,Silver Spring, MD.

Olinger, K. 1997. Sediment quality assessment for the tidal Christina River basin. Volume I:Report of findings. Delaware Department of Natural Resources and EnvironmentalControl, New Castle, DE.

Pinkney, A.E., D.W. Sutherland, R.E. Foley, and J.C. Harshbarger. 1995. Investigation ofsediment contamination and fish pathology at Featherstone National Wildlife Refuge,Virginia. U.S. Fish and Wildlife Service, Annapolis, Maryland. CBFO-C95-02.

Pinkney, A.E., J.C. Harshbarger, E.B. May, and M.J. Melancon. 2001. Tumor prevalence andbiomarkers of exposure in brown bullheads (Ameiurus nebulosm) from the TidalPotomac River Watershed. Environ. Toxicol. Chem. 20:1196-1205.

Pinkney, A.E., J.C. Harshbarger, E.B. May, and W.L. Reichert. 2002. Tumor prevalence andbiomarkers of exposure and response in brown bullheads (Ameiurus nebulosus) from theAnacostia River, Washington, D.C. and Tuckahoe River, Maryland. U.S. Fish and WildlifeService, Chesapeake Bay Field Office, Annapolis, MD. CBFO-C02-07.

Stokes, M.E., C.S. Davies, and G.G. Kock. 1995. Categorical Data Analysis Using the SASSystem. SAS Institute, Inc., Gary, North Carolina.

Vogelbein, W.G., J.W. Fournie, P.A. Van Veld, and R.J. Huggett. 1990. Hepatic neoplasms inthe mummichog Fundulus heteroclitus from a creosote-contaminated site. Cancer Res. 50:5978-5986.

Whitehead, A.C. 1995. Freshwater marsh kill if ish. In: L.E. Dove and R.M. Nyman, editors.Living Resources of the Delaware Estuary. The Delaware Estuary Program, pp. 205-210.

Woodward-Clyde Consultants, Inc. 1992. Risk assessment DuPont Newport Site Newport,Delaware August 7, 1992. Volume 2 of 2 Environmental Evaluation. Woodward-Clyde,Plymouth Meeting, PA.

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Woodward-Clyde Consultants, Inc. 1996. Phase III RI Scope of Work Former KoppersCompany, Inc., Newport Site. Woodward-Clyde, Blue Bell, PA.

(J:'.C'ONTAM proposals Jelltumsurv2003.wpd

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Alfred E. Pinkney, Aquatic Contaminants Team Leader

U.S. Fish and Wildlife Service, Chesapeake Bay Field Office, 177 Admiral Cochrane Drive,Annapolis, MD 21401. Tel. 410-573-4519; Fax 410-269-0832;e-mail: FRED_PINKNEY@FWS.GOVEDUCATION:Ph.D., Marine-Estuarine-Environmental Science, University of Maryland, College Park, MD,1988 (Research conducted at Chesapeake Biological Laboratory, Solomons, MD)M.S., Environmental Health Sciences, New York University, Tuxedo, NY, 1982B.S., Zoology, University of Michigan, Ann Arbor, MI, 1976FIELDS OF COMPETENCE AND EXPERIENCE:Aquatic toxicology, fish and invertebrate physiology and biochemistry, contaminant monitoring,ecological risk assessment, natural resource damage assessment.EMPLOYMENT:1993-present: Senior Environmental Contaminants Specialist/Team Leader, U.S. Fish andWildlife Service, Chesapeake Bay Field Office, Annapolis, Maryland1989-1993: Scientist, Versar, Inc., Columbia, Maryland1988-1989: Senior Scientist, ICF Inc., Fairfax, Virginia1986-1988: Graduate Research Assistant, Chesapeake Biological Laboratory, Solomons,Maryland1983-1985: Research Assistant, Johns Hopkins University, Applied Physics Laboratory, ShadySide, MarylandPRIMARY RESPONSIBILITIES:Develops and conducts studies assessing the impacts of contaminants on fish and wildl iferesources and their habitats; serves on the EPA Region III , Biological and Technical AssistanceGroup (BTAG), which reviews and advises EPA on ecological issues at hazardous waste sites.SELECTED PUBLICATIONS: (-20 peer-reviewed articles, 40 reports, 40 abstracts)Pinkney et al. 2001. Tumor prevalence and biomarkers of exposure in brown bullheads (Ameiurus nehulosus) from the tidal Potomac River watershed. Environ. Toxicol. Chem.20:1196-1205.Pinkney et al. 2000. Effects of the mosquito larvicides, temephos and methoprene, on insectpopulations in experimental ponds. Environ. Toxicol. Chem. 19:678-684.Pinkney et al. 1999. Effects of temephos (Abate 4E) on fiddler crabs (Uca pugnax and Ucaminax) on a Delaware salt marsh. J. Am. Mosq. Cont. Assoc. 15:321-329.Pinkney et al. 1999. Ecological risk assessment-Quantico embayment. Marine Corps CombatDevelopment Command - Quantico. Quantico, Virginia. CBFO-C99-03.Pinkney et al. 1998. Nontarget effects of the mosquito larvicides, temephos and methoprene, atBombay Hook and Prime Hook National Wildlife Refuges. USFWS, CBFO-98-01.Pinkney et al. 1997. Bioaccumulation of organochlorine contaminants in the Potomac River nearthe Old Landfill , Marine Corps Combat Development Command, Quantico, Virginia. FinalReport. USFWS, CBFO-C96-04.Pinkney et al. 1995. Sediment PAHs and tumors in brown bullhead (Ameiurus nehulosus) atFeatherstone National Wildlife Refuge, Virginia. USFWS, CBFO-C95-02.

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CURRICULUM VITAE

Name: John Carl Harshbarger, Ph.D.Telephone/Fax/Email: (202) 994-1056; (202) 994-2618; patjch@gwumc.edu

Education:1957 - B.A., Bridgewater College, Bridgewater, Virginia1959 - M.S., Virginia Polytechnic Institute, Blacksburg, Virginia1962 - Ph.D., Rutgers University, New Brunswick, New Jersey1966-88 - Postgraduate pathobiology courses: Harvard Univ. Medical School, Georgetown

Univ. Medical School and the NIH Graduate SchoolEmployment:

1962-64 - National Science Foundation Postdoctoral Fellowship in Insect Pathology, InsectPathology Laboratory, USDA, Beltsville, MD

1964-67 - Research Pathobiologist, University of California, Irvine, CA1967-95 - Director, Registry of Tumors in Lower Animals,

Smithsonian Institution, Washington, D.C.1995- pres. Professor of Pathology and Director, Registry of Tumors in Lower Animals,

George Washington University Medical Center, Washington, D.C.

Selected Bibliography:Baumann, P. C. and Harshbarger, J. C.: Long term trends in liver neoplasm epi/ootics of brownbullhead in the Black River, Ohio. Environ. Monit. Assess. 53: 213-223. 1998.Harshbarger, J. C., Chang, S. C., DeLanney, L. E., Rose, F. L., and Green, D. E.: Cutaneousmastocytomas in the neotenic caudate amphibians, Amhystoma tnexicanum (axolotl) andAmbystoma tigrinum (tiger salamander). J. Cancer Res. Clin. Oncol. 125(3/4): 187-192. 1999.Coffey, M., Phillips, K., Berg, C., Harshbarger, J., and Gross, K.: Middle Mississippi Riversturgeon health assessment: Illinois, Iowa and Missouri. U.S. Fish and Wildlife Service RegionalIdentification Number 3F25, Division Identification Number 9730003, Draft Report - February16,2000.Beckwith, L. G., Moore, J. L., Tsao-Wu, G. S., Harshbarger, J. C., and Cheng, K. C.:Ethylnitrosourea induces neoplasia in zebrafish (Danio rerio). Lab. Invest. 80(3): 379-385 +cover + editorial. 2000.Pinkney, A. E., Harshbarger, J. C., May, E. B., and Melancon, M. J..: Tumor prevalence andbiomarkers of exposure in brown bullheads (Ameiurus nehulosus) from the tidal Potomac Riverwatershed. Environ. Toxicol. Chem. 20:1196-1205. 2001.

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Chris_Guy@fws.gov

06/24/2003 02:28 PM

To: Matthew Mellon/R3/USEPAAJS@EPAcc:

Subject: proposal—delaware mummichogs

Christopher P. Guy177 Admiral Cochrane DriveAnnapolis MD 21401410-573-4529chris_guy3fws.gov

Forwarded by Chris Guy/CBFO/R5/FWS/DOI on 06/24/2003 02:20 PM

Fred Pinkney

Guy/CBFO/R5/FWS/DOIOFWS06/24/2003 02:14PM

mummichogs

To:

CC :

Chris

Subject: proposal --delaware

(See attached file: delltumsurv2003.wpd)

delltumsurv2003.wp<

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