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CRD 04-13 Ninth Flatfish Biology Conference, December 1-2, 2004, Water's Edge Resort, Westbrook, Connecticut

Abstracts - Poster Presentations


Winter Flounder, Pseudopleuronectes americanus,
Hatching Success as a Function of Burial Depth in the Laboratory

Walter J. Berry1, Elizabeth K. Hinchey1, Norman I. Rubinstein1, and Grace Klein-MacPhee2

1U.S. Environmental Protection Agency, Office of Research and Development, National Health and Environmental Effects Research Laboratory, Atlantic Ecology Division, Narragansett, RI 02882
2University of Rhode Island, Graduate School of Oceanography, Narragansett, RI 02882

Previous experiments have shown that viable hatch of winter flounder eggs is reduced when the eggs are buried by as little as one half of one egg diameter (approximately 0.5 mm of sediment).  This sensitivity to burial has resulted in seasonal banning of dredging in several northeastern U.S. estuaries.  In this study, a series of experiments was performed to better determine burial effects on hatching success of winter flounder.  In the first experiment, eggs were exposed to clean, fine-grained sediment with burial depths including a no-sediment control, dusting (< 0.5 mm), and up to 2 egg diameters (2 mm) of sediment.  A trend of decreased hatch success and delayed hatch date with increasing depth of burial relative to controls was observed: however, differences were not statistically significant (p>0.05).  In a second experiment, treatments included a no-sediment control, 4 egg diameters (4 mm) of clean, fine grained sediment, and between 0.5  to 6 diameters (0.5 - 6 mm) of highly contaminated, fine-grained sediment.  Eggs buried in 4 diameters of clean sediment did not hatch.  Hatch from eggs dusted with contaminated sediment was similar to controls, while eggs buried by 3 diameters of contaminated sediment had little or no hatching success. There was no evidence of delayed hatch.  The results of the second experiment must be interpreted with caution, however, due to the low control survival (21%).  Overall, the results of our initial trials indicate that winter flounder eggs may be more resistant to burial than previously thought. 


Effect of Contaminated Sediments on Mortality
and Predation Avoidance in Winter Flounder

Andrew F. J. Draxler1, Daniel Wieczorek1, Lori Davias2, Michael Schafer1, Yan Waguespack3, and Bashir Balogun3

1NMFS/ NEFSC/Howard Marine Laboratory, Highlands, NJ 07732
2North Carolina State University, Raleigh, NC 27695
3University of Maryland Eastern Shore, Department of Natural Sciences, Princess Anne, MD 21853

Young-of-the-year (YOY) winter flounder (Pseudopleuronectes americanus) held on sediment from the south branch of the Elizabeth River, VA, sustained significantly higher rates of in situ mortality and higher predation by bay shrimp (Crangon septemspinosa) than did fish held on less contaminated York River, VA sediment.  The mortality pattern paralleled previous toxicity results for winter flounder on Newark Bay, NJ sediment.  We speculate that observed effects on these juvenile demersal fish were the result of transdermally absorbed sediment components, most likely PAHs, which are well documented in the Elizabeth River.  Preliminary measurements show that respiration was lower in fish held on Elizabeth River sediment, which suggests a mechanism for the observed reduction in escape capability.


A Comparison of the Use of Estuarine Habitats by Flatfishes in South Carolina and Georgia

Guy’ DuBeck and Mary Carla Curran

Savannah State University, Department of Natural Sciences and Mathematics, P.O. Box 20600, Savannah, GA  31404

As part of a class project in fall 2004, we began a monitoring program to assess the abundance of flatfishes in a small tidal creek in South Carolina and one in Georgia.  The purpose of the exercise was to familiarize a student with various fishing gear in two marsh creeks - one accessible by boat and the other by foot.  In Country Club Creek, GA we conducted three replicate hauls with a 1-meter beam trawl (3-mm mesh).  In Chowan Creek, SC we conducted 3 replicate hauls with a 3-meter seine net.  The Georgia site was dominated by the blackcheek tonguefish Symphurus plagiusa, although we also obtained the summer flounder, Paralichthys dentatus.  We caught fewer flatfishes in South Carolina, possibly due to the gear type, but S. plagiusa and P. dentatus were present.  From historical summer data collected from 1998 –2001, fringed flounder, Etropus crossotus, and bay whiff, Citharichthys spilopterus, were also found at this site.  Future research will include comparing the efficiency of the seine and beam trawl in Chowan Creek.


Bay Whiff Distribution and Abundance in Galveston Bay, Texas

Lindsay Ann Glass

Texas A&M, 5007 Ave U #102, Galveston, TX 77551

Bay whiff (Citharichtys spilopterus) are the numerically dominant flatfish in the Galveston Bay estuary, yet little is know about the ecological significance of the species. Our objective was to characterize the distribution and abundance of bay whiff in Galveston Bay.  We used a one-meter beam trawl to sample three types of habitat (marsh edge, 1m depth water, and 2-to-5m water depth) in three main regions of the bay. Total length ranged between 20 to 30mm in the February, increasing to 40 to50mm in May. The highest density (0.618/m2) was measured in deep water habitat during March, and overall equal densities (0.579/m2) were measured in both east and west bay regions with lower densities (0.3/m2) in the central portion of Galveston Bay. Based on preliminary results, the large numbers of bay whiff in Galveston Bay could represent competition for resources with other flatfish of the same size range. 


The Abundance of Flatfishes in Country Club Creek, GA

Karen Harris and Mary Carla Curran

Savannah State University, Department of Natural Sciences and Mathematics, P.O. Box 20600, Savannah, GA  31404

The purpose of this study was to provide a quantitative comparison of the relative abundance and sizes of the flatfish species occupying a small estuarine tributary over the course of two seasons (Jan-Apr 2004) as part of an undergraduate research project.  Using three replicate hauls with a 1-meter beam trawl (3-mm mesh), we noted that the catch was dominated by the blackcheek tonguefish Symphurus plagiusa and, towards late spring, summer flounder Paralichthys dentatusS. plagiusa ranged from 5-20 individuals per date and lengths varied from 1.7-7.8 cm TL.  P. dentatus ranged from 0-23 individuals per date and lengths varied from 2.0-7.9 cm TL.  The southern flounder, Paralichthys lethostigma, and oscellated flounder, Ancylopsetta quadrocellata, were also collected.  This project will provide some preliminary data that may be used as part of a future long-term monitoring study. 


Differential Effects of Preservative Methods
on American Plaice and Atlantic Cod Ovarian Tissue

Nikolai Klibansky and Francis Juanes

University of Massachusetts, Department of Natural Resources Conservation, Amherst, MA 01003

Addition of relative fecundity estimates to stock-recruit models can significantly strengthen recruitment predictions, but confusion in the literature about the effects of common preservatives on ovarian tissue hinders the standardization of fecundity methods and comparison of fecundity data across studies.  In this research subsamples (~1.5g) of ovarian tissue of developing American plaice (Hippoglossoides platessoides) and Atlantic cod (Gadus morhua) were subjected to four preservative treatments (10% phosphate buffered formalin, non-toxic Gilsons solution, 70% ETOH, and freezing) to quantify effect on sample weight, and assess whether the preserved samples were suitable for measurement using digital image analysis.  All treatments preserved eggs well enough to measure digitally, though only eggs in formalin could be analyzed without first removing debris.  Mean weight of American plaice samples in the ethanol treatment increased by 25%, samples in 10% formalin increased by 20%, and samples in non-toxic Gilson’s solution decreased by 21% on average.  Samples that were frozen in distilled water increased by an average of 8%, but this change was not significantly different from zero.  Percent change in weight due to treatment differed significantly between the two species for all treatments except formalin.  In both species, coefficient of variation for the change in weight was highest for the frozen samples.  Ethanol altered the weight of the cod samples the least, while freezing altered the weight of plaice samples least, and formalin was the most stable treatment overall.  This information is useful in planning fecundity lab methods and comparing data across studies using different preservatives.


Use of Shallow Habitats by Juvenile Winter Flounder,
Pseudopleuronectes americanus,
along the Maine Coast

Mark A. Lazzari

Maine Department of Marine Resources, West Boothbay Harbor, ME 04575

Historically, the fisheries in Maine's shallow waters (< 10 m) provided resources of recreational and commercial importance, but no comprehensive studies of the fishes occupying these habitats along the entire coast ever occurred. Recent legislation has emphasized the importance of “essential fish habitat” including “those waters necessary for spawning, breeding, feeding, or growth” to healthy fisheries. 

We divided the entire Maine coast into three zones for sampling over five consecutive years. In 2000, we sampled the midcoast from Casco to Penobscot Bay, during 2001 - 2002 along the southern Maine coast to New Hampshire and along the eastern Maine coast to Canada in 2003 - 2004. Sampling with a 2.0-meter beam trawl (5 minute tows) provides estimates of the importance of various shallow water habitats (eelgrass, kelp, drift algae, sand/mud) to the early life history of fishes.

Winter flounder were a major component of midcoast estuaries (> 2 fish tow –1), but much less common to the east and south (< 0.2 tow –1). All shallow habitats function as nurseries. CPUE was consistent (0.73 – 1.0 tow –1) in the three major habitats sampled in eelgrass, Zostera marina, kelp, Laminaria longicruris, and over sand, but much lower in drift algae, Gracilaria sp. (0.10 tow –1). CPUE increases from 0.01 tow –1 in April to average about 1 tow –1 during June to September before dropping by half in October. Winter flounder ranged from one – 42 cm TL with most between 2 – 10 cm (73%) and few adults greater than 20 cm.


Impacts of Size on Growth and Early Maturity
in Female Yellowtail Flounder, Limanda ferruginea (Storer)

Anthony J. Manning, Margaret P.M. Burton and Laurence W. Crim

Ocean Sciences Centre, Memorial University of Newfoundland, St. John’s, Newfoundland, A1C 5S7 Canada

Yellowtail flounder is a small, early-maturing flatfish that has been considered as a candidate species for cold-water aquaculture in Newfoundland. The experiment examined three size classes of one-year-old, cultured female fish over a 14-month period. Small, medium and large females were separated as 0+ and young 1+ year fish. Pubertal development in 1+ fish was lowest, intermediate and highest for the small, large and medium size classes, respectively. Growth rates were highest for the small size class and lowest for the medium size class. Large fish still grew well but mature individuals of this group grew more slowly than immature individuals. These results reinforce the negative relationship between maturity and growth rate in flatfish. However, pubertal onset at one year of age was not always connected with a larger size in these cultured females.  


Real-time PCR Analysis of Steroid Enzyme Gene Expression During Atlantic Halibut (Hippoglossus hippoglossus) Development

Makoto Matsuoka1, Solveig van Nes2, Øivind Andersen2, Tillmann Benfey3, and Michael Reith1

1National Research Council of Canada, Institute for Marine Biosciences, Halifax, Nova Scotia, Canada B3H 3Z1
2Norwegian Institute of Aquaculture Research, Aas, Norway
3University of New Brunswick, Department of Biology, Fredericton, New Brunswick, Canada E3B 6E1

Atlantic halibut aquaculture would benefit from the development of all female lines, since females grow faster and reach a larger size than males.  Determining the period when gonad development occurs would define the critical time when hormonal or environmental treatments should be applied to influence sex ratios.  We investigated the transcription levels of the steroid hormone biosynthetic enzyme genes, aromatase (both brain- and ovary-type) and 11-beta hydroxylase, which catalyze key steps in the synthesis of 17 beta-estradiol and 11-ketotestosterone, respectively, in juveniles (16 - 67 mm in standard length) using real-time PCR.  The results suggest that treatments to manipulate sex ratios should be started by the time fish reach as small as 16 mm when both of the aromatase genes showed elevated levels in the brain.  For all three genes, elevations in expression levels were observed in both brain and gonad regions in fish prior to histological sex differentiation (approximately 32 mm).  The elevations in brain occurred slightly earlier than in gonad, supporting the idea that steroid hormone expression in the brain is a key determinant of phenotypic sex in fish.  However, we were unable to detect consistent differences in the expression patterns of these genes that would be diagnostic of sex.


PLEUROGENE: Genomics for the Enhancement
of Commercial Production of Atlantic Halibut and Senegal Sole

Makoto Matsuoka1, Michael Reith1, Joan Cerdà2, Harry M. Murray1,
Debbie Martin-Robichaud3, Brian Blanchard4, and Susan E. Douglas1

1Institute for Marine Biosciences, 1411 Oxford Street, Halifax, Nova Scotia, Canada B3H 3Z1
2Institute de Ciències del Mar, Consejo Superior de Investigaciones Científicas (CSIC), Passeig, Marítim 37-49, 08003-Barcelona, Spain
3Fisheries and Oceans Canada, Biological Station, 531 Brandy Cove Rd., St. Andrews, New Brunswick, Canada E5B 2L9
4Scotian Halibut Limited, P.O. Box 119, Clarks Harbour, Nova Scotia, Canada B0W 1P0

Atlantic halibut (Hippoglossus hippoglossus) and Senegal sole (Solea senegalensis) are two flatfishes yielding high value market products with good potential for aquaculture in eastern North America and Mediterranean Europe, respectively.  Production–related problems in these two evolutionary–related species may be addressed with improved knowledge of important basic biological processes such as reproduction, development, nutrition, genetics and immunity.  The use of genomic approaches to thoroughly characterize these processes will translate into knowledge that can be used to overcome the production obstacles and create (for PLEUROGENE is a new initiative funded through Genome Canada-Genome Spain for three years (2004-2007). There are two main goals: (i) the construction of genetic linkage maps Atlantic halibut and Senegal sole for use in the selection of improved broodstock based on molecular markers, and (ii) design, construction a flatfish microarray studies gene expression these two species.  High-throughput genome- proteome-based technologies will be used identification, characterization mapping genes important reproduction, larval development, immunity nutrition.  All the genetic and molecular information obtained in this project will be integrated into an interactive bioinformatic platform specifically developed for the project. The knowledge generated by the PLEUROGENE project will ultimately lead to the establishment of new technologies for the control of reproduction and optimization of larval health and nutrition in the Senegal sole, Atlantic halibut, and other related flatfish species under intensive culture conditions.


Effects of Contaminants on Winter Flounder Living
in a Sewage-Impacted Estuary-Jamaica Bay, NY

Lourdes Mena1, Lucia Cepriano2, Nancy Denslow3, Martin Schreibman4, and Anne E. McElroy1

1Stony Brook University/MSRC, Stony Brook, NY 11794
2State University of New York, Farmingdale, NY 11735
3University of Florida, Gainesville, FLA 32611
4Aquatic Research & Environmental Assessment Center, Brooklyn, NY 11210

Jamaica Bay, NY is an urban estuary that receives millions of gallons of sewage effluent daily making sewage effluent its primary source of freshwater. Extremely high levels environmental estrogen mimics (e.g., nonylphenol, estradiol, estrone) observed in sediments led us to postulate that benthic fish residing in Jamaica Bay, NY are likely targets for endocrine disruption and, potentially, reproductive impairment. We collected adult, juvenile and young-of-the-year winter flounder, Pseudopleuronectes americanus, from multiple sites in Jamaica Bay and a reference site off the east coast of Long Island, Shinnecock Bay, in the spring of 2002 and 2003.  Very few male fish were found at the site with the highest levels of endocrine disruptors. Levels of circulating 17 beta-estradiol (E2), vitellogenin (VTG) and 11-Ketotestosterone (11-KT) showed unusual patterns of endocrine disruption in this species. Females from the most contaminated site showed significantly higher levels of VTG and males only showed decreased levels of E2 and 11-KT as compared to reference fish. Histological analysis of liver and gonad tissue are also ongoing.


Habitat for Flatfish: The Relative Importance of Depth, Temperature and Substrate

Elizabeth T. Methratta, Brian E. Smith, and Jason S. Link

NMFS/NEFSC/Woods Hole Laboratory, 166 Water Street, Woods Hole, MA 02543

Understanding how flatfish abundance, distribution, and production are related to habitat factors and how these relationships change across ontogeny at large spatial scales are key components to the development of spatial tools for fisheries management.  A geographic information system (GIS) was used to bin USGS substrate grain size data and a 34-year NEFSC time series of environmental and biological data into 10-minute squares.  Data were averaged over 2 seasons (fall and spring) for each of 3 time blocks (1968-79, 1980-89 and 1990-02).  Multivariate statistical analyses were then used to examine the relative importance of bottom temperature, depth, substrate grain size, season, and time block for individual size classes for four flatfish species: American plaice (Hippoglossoides platessoides), fourspot flounder (Paralichthys oblongus), winter flounder (Pseudopleuronectes americanus), and yellowtail flounder (Limanda ferruginea).  Depth explained the largest proportion of variance in the distribution of size classes for American plaice, winter flounder, and fourspot flounder with larger individuals more abundant in deeper waters.  This pattern reflects the characteristic onshore-offshore spawning migration undertaken by adults of these species.  For yellowtail flounder, which inhabits deeper waters, time block was the strongest explanatory factor, with larger individuals relatively more abundant later in the time series.  This is likely a response to the combined effects of changing exploitation and subsequent recruitment patterns.  In contrast, larger individuals declined over the time series for the other three species most likely in response to the preferential exploitation of bigger fish.  Substrate grain size explained 2-13% of the explainable variance in species size-class distributions.  Larger substrates were associated with shallower depths and smaller individuals for all four species.  Although more refined estimates of essential fish habitat for marine fishes will continue to be challenging, novel geostatistical approaches offer strong potential in meeting these challenges. 


Development of Digestive Capacity in Larval Atlantic Halibut: A Preliminary Survey

Harry M. Murray, Jeffrey W. Gallant, Stewart C. Johnson, and Susan E. Douglas

Institute for Marine Biosciences, National Research Council of Canada, 1411 Oxford Street, Halifax, Nova Scotia B3H 3Z1

The objectives of the present study were to identify the location and timing of gene expression of specific digestive enzymes in first-feeding Atlantic halibut larvae and juveniles. Prior to the start of first feeding, the gastrointestinal tract (GIT) is divided into anterior, mid and hindgut regions. The liver is present at this time, as is the pancreas. During larval development the pancreas changes from a compact organ to a diffuse tissue interspersed though much of the mesentery surrounding the GIT. Functional gastric glands are not present until approximately 66 days post-hatch (dph).  Using primers based on winter flounder digestive enzyme gene sequences for bile salt-activated lipase (BAL), trypsinogen (Trp), and pepsinogen (Pep), we were able to amplify products from RNA extracted from whole larvae and juveniles using RT-PCR. These products were sequenced and the sequences used to design halibut gene-specific primers for the above enzymes. RT-PCR analysis using halibut-specific primers revealed that Trp and BAL gene expression was evident at least from the time of first feeding. Pep gene expression was not detectable until 80 dph. In situ hybridization with DIG-labeled antisense RNA probes localized expression of BAL and Trp to the exocrine pancreas. Pep expression was localized only to the glandular regions of the stomach. These data provide a first step toward understanding the molecular biology underlying the ontogeny of digestive capacity in Atlantic halibut.     


Fish Community Structure of Intertidal Habitats of the Mount Hope Bay Estuary with Emphasis on Juvenile Winter Flounder, Pseudopluronectes americanus

Adrienne Pappal

University of Massachusetts-Dartmouth School for Marine Science and Technology, 706 Rodney French Blvd, New Bedford, MA 02744

Fish respond to a variety of factors in the selection of habitat, including prey availability, predator avoidance, and competition. Fish are able to respond to these cues and alter their distribution to minimize metabolic cost. Identifying which habitats may be preferred for juvenile winter flounder is critical in conservation efforts and the definition of essential fish habitat for this species. My masters research focuses on the juvenile winter flounder population of Mount Hope Bay, where there is large areas of intertidal rocky/cobble habitat, with patches of Spartina patens. Intertidal habitat use of juvenile winter flounder was assessed using a three-pronged effort of field study, habitat mapping and laboratory study. Collections of winter flounder on site were attempted using a 50ft beach seine and a blocking tidal trap with mixed results. Intertidal habitats were quantified using random transects and .5m x .5m quadrats. Ongoing laboratory study will address in depth the substrate and vegetation preferences of juvenile winter flounder using habitat types present in the field. These laboratory studies will mimic the dominant rocky/cobble substrate and emergent Spartina patens grass present in Mount Hope Bay to determine use by juvenile winter flounder. These habitats are not sampled effectively with beach seines commonly used in monitoring programs, therefore juvenile winter flounder may be under sampled in Mount Hope Bay. It is generally understood in the literature that coarse/ sandy substrates or organic coves are the preferred habitat for juvenile winter flounder, but it is yet unclear if juvenile winter flounder utilize rocky/cobble substrates to any great extent.


Regulation of Carbonic Anhydrase-Dependent Renal Sulfate Secretion by Cortisol in Winter Flounder, Pseudopleuronectes americanus

Ryan M. Pelis1,2, James E. Goldmeyer1, Joseph Crivello1, and J. Larry Renfro1,2

1University of Connecticut, Department of Physiology and Neurobiology, Storrs, CT 06269-4156
 2Mount Desert Island Biological Laboratory, Salisbury Cove, ME 04672

Sulfate secretion by the marine teleost renal proximal tubule is stimulated by cortisol and dependent on carbonic anhydrase (CA).  Thus, cortisol modulation of sulfate secretion may be through stimulation of CA.  To study this relationship primary cultures of flounder renal proximal tubule epithelium (fPTCs) were exposed to elevated hydrocortisone (HC) or reduced HC for 5 days prior to determination of CA activity, CAII protein abundance (immunoblotting), and transepithelial transport of sulfate in Ussing chambers.  Degenerate primers and RT-PCR were used to obtain a partial cDNA clone (327 bp) of fPTC CAII that exhibited high sequence similarity to CAII from numerous species.  CAII was localized to the cytosol and plasma membranes of intact flounder renal tubules with immunohistochemistry.  Treatment of fPTCs with reduced HC caused reductions in CA activity (28%), CAII protein abundance (65%), and active sulfate secretion (28%), without affecting cell differentiation.  Treatment of fPTCs with methazolamide (MTHZ, 0.1 mM), a CA inhibitor, reduced active sulfate secretion 55%.  Treatment with reduced HC and MTHZ together had the same effect as MTHZ alone.  These data demonstrate that the marine teleost proximal tubule contains both cytosolic and membrane-associated CAII and that cortisol directly stimulates CA activity, CAII abundance, and a fraction of sulfate secretion that is CA-dependent.  Supported by NSF. 


A Flatfish Perspective on Asymmetry:
What Genes Mediate Southern Flounder Metamorphosis?

Alex M. Schreiber and Yan Tan

Carnegie Institution of Washington, Department of Embryology, 115 West University Pkwy, Baltimore, MD 21210

Metamorphosis in frogs and flatfish is mediated by thyroid hormone (TH).  The craniofacial morphology for both tadpoles and larvae is initially bilaterally symmetric, but during metamorphosis the skull and jaw undergo abrupt and extensive remodeling. In contrast to the newly remodeled froglet head that retains bilateral symmetry, the juvenile flatfish has metamorphosed into the world’s most asymmetric vertebrate: translocation of one eye to the opposite side of the head is accompanied by asymmetric skull and jaw development.  What TH-responsive genes mediate flatfish metamorphosis in general, and craniofacial remodeling in particular? Does asymmetric gene expression correspond with left and right sidedness within and among flatfish species? How evolutionarily conserved is metamorphosis among vertebrates?  The first step in addressing these questions is to identify genes that are differentially expressed during metamorphosis.  Premetamorphic 13-day old southern flounder were treated for 3 days with either TH or an inhibitor of TH production (methimazole). Messenger RNA from each group was purified, and differentially-expressed genes were enriched using a PCR-based subtractive hybridization procedure. Though we will sequence 3,000 differentially-expressed clones, here we describe the first 300 TH-upregulated and 160 downregulated clones of our screen based upon sequence homology with known vertebrate genes. Upregulated genes include several implicated in bone remodeling (periostin, PTH-responsive osteosarcoma protein, Sox, and alpha-2-HS-glycoprotein), as well as development of skin, gut, muscle, central nervous system, and cell proliferation. Downregulated genes were homologous to proteins expressed in skin (keratins), gut (trypsin, lipid binding protein), blood proteins (parvalbumin) and many metabolic proteins.


Benthic Food Webs on Georges Bank: Where Do Flatfish Fit into the Picture?

Brian. E. Smith, Elizabeth T. Methratta, and Jason S. Link

NMFS/NEFSC/Woods Hole Laboratory, 166 Water Street, Woods Hole, MA 02543

Flatfish are an important component of benthic food webs.  These species provide major links between the benthos and upper trophic level predators in the northeast US continental shelf ecosystem.  We examined more than 30 years of food habits data for seven species of flatfish and their major predators from Georges Bank to evaluate the temporal variation in flatfish predator-prey relationships.  The morphology of flatfish (gape width) suggests that most species have a strong preference for certain prey, adopting either a benthivorous or piscivorous diet after reaching adulthood.  Summer flounder (Paralichthys dentatus) and fourspot flounder (Paralichthys oblongus) were generally piscivorous, consuming herring, sand lance, hakes, mackerel, and similar forage fish.  American plaice (Hippoglossoides platessoides) specialized on ophiuroids and sand dollars (Echinarachniusparma).  Yellowtail flounder (Limanda ferruginea), winter flounder (Pseudopleuronectes americanus), windowpane (Scophthalmus aquosus), and witch flounder (Glyptocephalus cynoglossus)ate primarily polychaetes, gammarids and similar benthic invertebrates.  Significant shifts in diet were observed for summer and fourspot flounder across the time series.  In contrast, changes in diet composition were not apparent for the benthivorous flatfish.  The major predators of flatfish on Georges Bank include spiny dogfish, Atlantic cod, red hake, white hake, sea raven, goosefish, longhorn sculpin, winter skate, little skate, and piscivorous flatfish.  Our results suggest that the major predator-prey relationships in the Georges Bank benthic food web have not fundamentally changed over time, despite changing energy flows and abundance over the past three decades.


Elemental Fingerprints of Juvenile Winter Flounder Otoliths
from Narragansett Bay, RI and Surrounding Coastal Ponds

Brian K. Taplin1, Richard J. Pruell1, John Brazner2, Ross Kean3, and Jennifer Yordy4

1US EPA, ORD, NHEERL, Atlantic Ecology Division,  27 Tarzwell Drive, Narragansett, RI 02882
2US EPA, Mid-Continent Ecology Division,  6201 Congdon Blvd, Duluth, MN 55804
3Research and Productivity Council, 921 College Hill Road,  Fredericton, New Brunswick, Canada E3B 6Z9
4Medical University of South Carolina,  171 Ashley Avenue, Charleston, SC 29425

Elemental fingerprinting has become a powerful tool in fisheries science for identifying fish migration patterns, seasonal changes in habitat use, and for delineating the nursery origins of adult fish populations. In this study we investigated whether elemental signatures in juvenile winter flounder (Pseudopleuronectes americanus) otoliths, collected from different habitat types and locations, could be used as natal fingerprints.

Juvenile flounder were collected from different nearshore habitats (unvegetated, macroalgae, and eelgrass) and locations (upper, mid, lower) within Narragansett Bay, RI (USA) and surrounding coastal ponds. Sagittal otoliths were removed, cleaned under a laminar flow hood and digested with high purity nitric acid. Elemental analysis was then conducted using Inductively Coupled Plasma Mass Spectrometry (ICP-MS) and or Inductively Coupled Plasma Atomic Emission Spectroscopy (ICP-AES).

The concentrations of nine elements (Ca, Sr, K, Na, Ba, Rb, Li, Mg and Mn) were measured and statistically analyzed using multivariate techniques. Both canonical discriminant analysis (CDA) and linear discriminate function analysis (LDFA) showed that otolith chemical fingerprints differed between fish collected from Narragansett Bay and the coastal ponds.  In addition, significant differences were observed in the otolith chemistry of fish collected from the three habitat types (unvegetated, macroalgae, and eelgrass).  These differences were more pronounced for comparisons either within the Bay or the coastal ponds and less pronounced for the combined data including all stations. In order to enhance our ability to differentiate among juvenile nursery areas, additional measurements including stable carbon and oxygen isotopes and rare earth elements will be investigated in the future.


Fin Rot in Winter Flounder from New Haven Harbor, CT:
The Clinical Profile of a Wasting Disease 

John J. Ziskowski, Jose Pereira, Renee Mercaldo-Allen, and Catherine Kuropat

NMFS/NEFSC/Milford Laboratory, 212 Rogers Avenue, Milford, CT 06460

Fin rot is a wasting disease characterized by fin loss and muscular emaciation often found in winter flounder sampled from urban estuaries.  This condition was observed in nearly 50% of flounder present in trawl catches from New Haven Harbor, Connecticut during the 1980’s.   From 1988 to 1990, 106 fish were processed for blood serum analyses to gain an understanding of the underlying physiological effects of this disease condition.  Initially, blood was removed from affected fish and controls at sea, soon after capture.  About half-way through the study, it was decided to take advantage of the wet lab capabilities of the Milford Laboratory and return specimens to shore, holding them overnight for blood removal and processing the next day.  Fish were identified by sex, measured, fin loss was quantified, hematocrits determined and blood serum chemistry values measured for: hemoglobin, calcium, bilirubin, osmolality, phosphate, and total protein.  A categorical database was created using fin rot (1/0) as the outcome variable and all other clinical parameters as predictor variables.  Stepwise Logistic Regression was used to model the contribution of blood values resulting in the final fin rot outcome.  Statistical analyses were done separately for each sex and for each mode of sampling.  For flounder that were sampled after an overnight rest period, hematocrit and calcium  values were the primary predictors of fin rot in female fish only.  Hematocrits were depressed and calcium values were elevated in these female fin-rotted fish.  Hemoglobin levels were the only predictors of fin rot on female flounder sampled at sea; calcium levels were not identified as a contributing variable in the final Stepwise Regression   model. Only 6 male flounder were available for sampling at sea; an insufficient number for statistical analysis.  Blood parameter values varied for diseased fish, depending on the magnitude of fin loss.  The etiology of fin rot is not known but could result from toxic chemicals or microbes adversely impacting resident flounder through the food chain, resulting in anemia and hypercalcemia in affected individuals.


Axial Skeletal Anomalies in Commercially Important Fish from the North Atlantic

John Ziskowski

NMFS/NEFSC/Milford Laboratory, Milford, CT 06460

Deformities of the axial skeleton are usually observed in fish in the form of dwarfism, spinal curvature, and cranial compression.  When radiographs are displayed in fish with these conditions; the following underlying structural deformities can be seen: fusion of vertebral centra, complexed vertebrae resulting from incomplete fission during embryonic development, accessory spinal processes, reduced spinal processes, deformed spinal processes, reduced centra, and deformed centra.  Species displayed include: winter flounder, American plaice, yellowtail flounder, witch flounder, Dover sole from the North Sea, Acadian redfish, and alewife. Specimens were obtained from NMFS research vessels operating in the Gulf of Maine and Georges Bank, from Boston Harbor during New England Aquarium’s Fish Days, and from trawling activities conducted by the NMFS Milford Laboratory in New Haven Harbor, Connecticut.  The Dover sole was purchased at a fish market. Associated with this poster presentation, radiographs of fish will be displayed.